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Samara: the international newsletter of the Millennium Seed Bank Partnership

E-newsletter

Welcome to the Samara e-newsletter. The e-newsletter is published three times a year in March, June and September, starting September 2021. It aims to share up-to-date news and stories from across the Millennium Seed Bank Partnership (MSBP) as well as share information of relevance to the partnership.

The e-newsletter sits alongside the annual print version of Samara, published in December. More information and past issues of the print edition can be found on the dedicated Samara print edition webpage.

If you have any feedback or would like to get in touch with the Samara team, please email samara@kew.org .

Issue 4: September 2022

In this issue we hear about field adventures in Svalbard and lab work on the endemic Lobelioideae of Hawaiʻi. As well as steps towards creating the third edition of the Red Book of the Republic of Azerbaijan, the progress of seed banking at Bali Botanic Garden and the creation of a Seed Production Garden on Kangaroo Island, Australia. This issue's interview is with Dr Dian Latifah, Coordinator of Seed Conservation Research Group within the National Research and Innovation Agency (BRIN), Indonesia. The seed question of the quarter looks at the difference between mouldy and infested seeds when doing cut-tests, X-rays and germination testing.

A yellow flower with 5 petals each with a fringed margin
Nymphoides geminata, Kangaroo Island, Australia. (Photo: SASCC).
A tall woody stem with leaf scars, ending in several whorls of simple lanceolate leaves with nine visible infloresence spikes emerging from the top. On each infloresence spike is a row many curved pink flowers all facing towards the sky.
Trematolobelia kauaiensis, Hawaiʻi. (Photo: Ken Wood).
A close up photo of the fruiting stalk of alpine sorrel. The seeds are round and flat with red outline
Oxyria digyna, Svalbard. (Photo: RBG Kew).
A low growing herbaceous plant on a rocky substrate. The leaves are small, simple and pointed and are opposite each other on the stem. There are flower buds from the top of each stalk with a purple tinge to them. The flowers are white with a bifucated tip.
Cerastium latifolium, Azerbaijan. (Photo: Elman Yusifov).

Stories from the field and lab this issue explore the Lobelioideae of Hawaiʻi and seed collecting in Svalbard.

The remote archipelago of Hawaiʻi in the Pacific Ocean is a hotspot of endemism with nearly 90% of the native flora being endemic to the islands, many being single-island or even narrower endemics (Sakai et al., 2002). Of the nearly 1,400 native plant taxa, nearly half are considered species of conservation importance and more than 230 plant taxa survive with fewer than 50 individuals remaining in the wild. There are several reasons for this endangerment, including habitat destruction, loss of pollinators, invasive species (including plants, rodents, and diseases), and climate change.

A tall woody stem with leaf scars, ending in several whorls of simple lanceolate leaves with nine visible infloresence spikes emerging from the top. On each infloresence spike is a row many curved pink flowers all facing towards the sky. In the background are forested hills and the ocean
Trematolobelia kauaiensis in flower in its natural habitat on Kauaʻi. Endemic to the island of Kauaʻi, seeds were used in the studies done at the MSB. (Photo: Ken Wood).
A tall woody stem with with leaf scars, ending in several whorls of simple lanceolate leaves all curving downwards. The bottom leaves are brown. From the top of the plant multiple horizontal infloresence spikes are protruding. Each infloresence spike has many pink flowers each with many recurved pink petals and protruding anthers.
Trematolobelia kauaiensis in flower in its natural habitat on Kauaʻi. Endemic to the island of Kauaʻi, seeds were used in the studies done at the MSB. (Photo: Ken Wood).

A diversity of complementary conservation measures are being taken to safeguard these plants, from seed banking and ex situ collections of plants in botanical gardens, to propagation and in situ out-planting in their natural habitat with varying success. Seed banks are fundamental resources for plant conservation, simultaneously providing a safety net through longer-term storage, as well as a means for linking ex situ conservation with in situ restoration and reintroduction. However, seed storage behavior varies greatly among species (Hong & Ellis, 1996) and understanding seed behavior is critical for successful seed banking and conservation. There is therefore an urgent need to understand evolutionary and ecological relationships with morphological and physiological seed traits that determine seed storage behavior.

While the majority of species are thought to produce orthodox seeds (i.e., desiccation and freeze tolerant), an estimated 8% of plant species are recalcitrant, or sensitive to desiccation (Wyse & Dickie, 2017). With some key families, life forms, biomes and levels of extinction risk having higher levels of non-orthodox behavior. In these instances, the proportion of species with recalcitrant seeds could be 40-70%. For those species that produce more desiccation tolerant seeds, there is a gradient of storage behavior, which includes species with intermediate storage behavior which show desiccation tolerance but poor storage at -20°C, where apparently orthodox seeds show a more rapid than expected decrease in viability over the course of a decade (Walters, 2015). Fully orthodox seeds can be expected to survive dry, cold storage for many decades.

Efforts to predict storage behavior based on measurable morphological or physiological seed traits (e.g., seed mass, lipid phase state changes, moisture content, and dormancy class) have thus far had limited success. This is particularly true for physiological traits that have historically been challenging to measure (like lipid phase state changes), but for which new technology has made measurements more accessible. Efforts to understand how seed storage behavior relates to ecological and phylogenetic relationships have been limited. Research has shown that storage behavior of seeds can be predicted by habitat and habit (Tweddle et al., 2003; Wyse & Dickie, 2017) and that there is a phylogenetic signal (Dickie & Pritchard, 2002) indicating storage behavior may be relatively conserved at low taxonomic levels. Moreover, there are suggested physical co-correlants of desiccation (in)tolerance, including seed mass and seed coat: whole seed mass ratio (Daws et al., 2006).

The endemic Hawaiian Lobelioideae (Campanulaceae; 6 genera, 126 spp.) exemplify adaptive radiation in plants and comprise the largest family in the flora of the most isolated archipelago on Earth (Wagner et al., 2005). Endemic Campanulaceae is distributed on seven of the eight main Hawaiian Islands ranging in elevation from <100 to >4000 m a.s.l. and demonstrates remarkable diversity of habit (e.g., shrubs, trees, rosettes, succulents, vines, epiphytes) and habitat (e.g., high elevation bogs, cliff faces, forests; Wagner et al., 1999). While the majority of species in the family produce orthodox seeds (i.e., desiccation and freeze tolerant), this is not the case with Hawaiian-endemic Campanulaceae which appear to have anomalous responses to conventional seed storage methods (i.e., equilibrium to 15-25% relative humidity (RH), and -18°C storage; Chau et al., 2019). Consequently, Hawaiian lobeliads provide an ideal model group for studying prediction of seed behavior though comparison with Campanulaceae globally.

A straight green stem with many leaf scars. At the top of the stem a whorl of deep purple round fruits are present. From the top of the long stemed leaves are shooting. The leaves a simple with a tooth margin and a pointed tip.
Delissea rhytidosperma in fruit in the NTBG Conservation Nursery. Endemic to Kauaʻi, now extinct in the wild. (Photo: Seana Walsh).
Four small seeds each less than 1mm long. Each seed is a slightly different shape between round and oblong. The surface of the seeds are light brown appearing lightly wrinkled
Delissea rhytidosperma seeds collected in 2022 in NTBG's Limahuli Garden and Preserve. (Photo: Dustin Wolkis).

As Seed Bank and Laboratory Manager at the National Tropical Botanical Garden (NTBG) headquartered on Kauaʻi, Hawaiʻi and a PhD Student at the University of Copenhagen Natural History Museum of Denmark, I seek to understand factors that affect seed longevity in the native Hawaiian flora. I recently traveled from Hawaiʻi to the UK for a research stay at RBG Kew’s Millennium Seed Bank (MSB) to study lipid thermal properties in the Hawaiian lobelioids and related groups. I targeted one representative species from each of the six Hawaiian clades from a recent global Lobelioideae phylogeny (Kagame et al., 2021), and one species from a closely related clade from the Marquesas from the NTBG Seed Bank and Lab collection, as well as one species each from three other closely related clades from the MSB collection. Unfortunately, I was not able to bring species from one of the Hawaiian clades outside of the United States as all taxa in it are federally listed as endangered in the US and permits could not be obtained. Supervised by Drs. Hugh Pritchard, Louise Colville, and Daniel Ballesteros, I used differential scanning calorimetry to capture the lipid thermal fingerprints for three replicates of each species, as well as performed annealing experiments at two temperatures. Somewhat serendipitously, on the last day of my visit, I was able to collect seeds of the critically endangered (possibly extinct in the wild) Delissea rhytidosperma (Walsh, 2015) on site, from the MSB research greenhouse, thereby capturing the final clade in the study that otherwise would not have been possible. By investigating the interactions between temperature and lipid melt and crystallization behavior over warming, we hope to gain insight into why these species exhibit poor storage behavior during conventional seed bank storage.

Dustin is wearing a white lab coat, stood behind a bench with a microscope. He is using tweezers to pick up seeds from a black dish
Dustin preparing seed for analysis in the MSB seed cleaning laboratory. (Photo: Michael Way).
Dustin wearing a white lab coat, bent over a small black tray of seeds holding tweezers. Next to him on the bench is a microscope with a small black tray on ready for the next sample
Dustin preparing seed for analysis in the MSB cleaning laboratory. (Photo: Michael Way).
References
  • Chau, M.M., Chambers, T., Weisenberger, L., Keir, M., Kroessig, T.I., Wolkis, D., Kam, R. & Yoshinaga, A.Y. (2019) Seed freeze sensitivity and ex situ longevity of 295 species in the native Hawaiian flora. American Journal of Botany. 106: 1248-1270. DOI: https://doi.org/10.1002/ajb2.1351
  • Daws, M.I., Garwood, N.C. & Pritchard, H.W. (2006) Prediction of desiccation sensitivity in seeds of woody species: a probabilistic model based on two seed traits and 104 species. Annals of Botany. 97: 667-674. DOI: https://doi.org/10.1093/aob/mcl1022
  • Dickie, J.B. & Pritchard, H.W. (2002) Systematic and Evolutionary Aspects of Desiccation Tolerance in Seeds. In: Desiccation and Survival in Plants: drying without dying. Eds. Black, M. & Pritchard H.W. CABI publishing.
  • Hong, R.H. & Ellis, T.D. (1996) A protocol to determine seed storage behaviour. IPGRI Technical Bulletin No 1. Rome Italy: International Plant Genetic Resources Institute.
  • Kagame, S.P., Gichira, A.W., Chen, L., & Wang, Q. (2021) Systematics of Lobelioideae (Campanulaceae): review, phylogenetic and biogeographic analyses. PhytoKeys. 174: 13–45. DOI: https://doi.org/10.3897/phytokeys.174.59555
  • Sakai, A.K., Wagner, W.L. & Mehrhoff, L.A. (2002) Patterns of endangerment in the Hawaiian flora. Systematic Biology. 51: 276-302. DOI: https://doi.org/10.1080/10635150252899770
  • Tweddle, J.C., Dickie, J.B., Baskin, C.C. & Baskin, J.M. (2003) Ecological aspects of seed desiccation sensitivity. Journal of Ecology. 91: 294-304. DOI: https://doi.org/10.1046/j.1365-2745.2003.00760
  • Wagner, W.L., Herbst, D.R. & Lorence, D.H. (2005) Flora of the Hawaiian Islands website
  • Wagner, W.L., Herbst, D.R., Sohmer, S.H. & Wilson-Ramsey, Y. (1999) Manual of flowering plants of Hawai’i. University of Hawai’i Press.
  • Walsh, A. (2015) Delissea rhytidosperma. The IUCN Red List of Threatened Species 2015. e.T7986283. DOI: https://dx.doi.org/10.2305/IUCN.UK.2015-4.RLTS.T79862837A79862844.en
  • Walters, C. (2015) Genebanking seeds from natural populations. Natural Areas Journal. 35: 98-105. DOI: https://doi.org/10.3375/043.035.0114
  • Wyse, S.V. & Dickie, J.B. (2017) Predicting the global incidence of seed desiccation sensitivity. Journal of Ecology. 105: 1082-1093. DOI: https://doi.org/10.1111/1365-2745.12725

We have recently returned from the arctic, where we were carrying out the first year of fieldwork for a three-year project to conserve arctic flora. Funded by the Marris-Webbe Charitable Trust, the project will see seed collections made in three arctic locations – Svalbard (Norway), Disko Island (Greenland) and the Latnjajaure Lake area (Sweden). Research into the adaptability of tundra vegetation to climate change will be conducted, in addition to research looking at how resulting species and community level changes influence the functioning of arctic ecosystems. The Millennium Seed Bank (MSB) team will focus on the seed conservation aspects of the project, while Dr Anne Bjorkman’s team from the University of Gothenburg, Sweden will undertake the research and provide support with field sites and logistics.

Spitsbergen, the largest island in Svalbard, is not your typical summer destination – at 78°N, with forecast highs of 5°C, meltwater leading to boggy conditions and the promise of 24-hour sunlight, it sounded a long way from the heatwaves which enveloped Europe this year. The other fact that makes it stand out is that you cannot leave the main settlement of Longyearbyen (the world’s northernmost settlement) without a rifle and flares – to keep polar bears at bay.

The back of a lady who is carrying a backpack and also has a rifle over her shoulder
Safety in Svalbard. (Photo: RBG Kew).

The archipelago of Svalbard was completely ice-bound until 10,000 years ago, meaning that the vascular plant species found there have migrated relatively recently from Greenland, America, and Russia. The west coast of Spitsbergen (where the fieldwork took place) has a particularly favourable climate despite its latitude, as a result of the warm North Atlantic current.

A deep glacial valley with sparse low level vegetation and a rocky substrate. Two people are visible bending down looking at vegetation
Endolen to the east of Longyearbyen - our first collecting site. (Photo: RBG Kew).
A clump of short plants with flowering stems a couple of centimetres off the ground amongst a rocky substrate in a glacial valley. Two seed collectors are visible in the background
Bjørndale to the west of Longyearbyen to Dryas octopetala. (Photo: RBG Kew).

Luckily it had been an early spring this year, meaning that seed set was good when we arrived at the end of July. Over eight days we managed to make 20 collections from the area surrounding Longyearbyen. These included the Svalbard Poppy (Papaver dahlianum), Arctic Cottongrass (Eriophorum scheuchzeri), and the Polar Willow (Salix polaris). The latter is one of only two ‘trees’ to occur in Svalbard, and has a low, creeping form less than 5cm tall – quite different from most Willow trees we know in the UK! Everything we collected was less than 20cm tall, which meant a lot of bending down and kneeling, and our main competition for the seeds seemed to be the hungry reindeer and geese that we often found nearby. Having explored higher up the valleys towards the glaciers and right down the coast of the fjord where beluga whales could be spotted, we found that most of the seeds were (not surprisingly) found in the valley bottoms and lower slopes, and along the coast. The fewer plants found growing higher up had not yet had time for their seeds to ripen due to later snow melt and cooler temperatures.

Rocky ground substrate with two different small plants, the one on the left has lighter green leaves and fruiting stems a couple of centimetres tall. The one on the right has brighter green leaves and the fruiting stems are much closer to the ground.
Two Saxifraga species growing together (Saxifraga cespitosa left, and Saxifraga oppositifolia right). (Photo: RBG Kew).
A close up photo of the fruiting stalk of alpine sorrel. The seeds are round and flat with red outline
Alpine sorrel, Oxyria digyna. (Photo: RBG Kew).

Our collections, and their accompanying herbarium specimens will soon arrive back in the UK for us to process and store. Half of each seed collection and an herbarium specimen will be kept at the National Seed Bank of Norway, in the Botanical Garden in Oslo, to ensure the safety of the collections through duplication.

3 people standing in a row smiling behind a work bench. The work bench has a variety of paper envelopes and cloth bags on it along with a stack of herbarium specimens
Collections and vouchers safely delivered in Linn, Kristina and Tor at the National Seed Bank of Norway, Oslo. (Photo: RBG Kew).

Across the MSBP articles this issue come from Azerbaijan, Indonesia and Australia.

A far reaching view of lakes and mountains with a foreground close-up of specimens of purple and yellow flowers growing amongst lush grass on a steep incline
Goygol Lake, the Lesser Caucasus Mountains. (Photo: Fuad Guliyev).

The wild flora of Azerbaijan is extremely rich and diverse, with more than 5,000 species of higher plants. Azerbaijan's natural vegetation and its flora are exposed to anthropogenic influences, and the number of valuable plant species is decreasing, with some now threatened with extinction. This is gradually having a negative effect on the ecosystem. Our latest research shows that at least 10% of the higher plants of Azerbaijan are rare and endangered. There are many relicts as well as endemic plants of the Caucasus and Azerbaijan which are collected from their natural habitat for use in the production of food, fodder, medicine, spices or decorative items and have thus become rare. To address this, in recent years, the Institute of Botany has been carrying out research on the ex situ protection of endangered plants, as well as developing the intraspecies systematics reflecting the gene pool of those species; studying limiting factors under in situ conditions; including investigating the reasons for the increase of the limiting factors, and ecological assessments. Ecosystem management requires a comprehensive strategic approach, which means studying and protecting the coexistence of land, water, and wildlife, in other words, all living systems.

A flat-topped rock formation covered with lichen, water pools and a bubbling geyser
Istisu, Kalbajar district, Karabakh. (Photo: Elman Yusifov).

An important factor for the detection and documentation of rare and endangered plants is the preparation of Red Books and Red Lists. In 2013, the 2nd edition of the Red Book of the Republic of Azerbaijan (300 plant species) was published. The results of scientific research show that the number of plants to be included on this list may now have doubled. To help identify the species that need protection, monitoring and expeditions are organised in different regions of the country based on lists compiled by Azerbaijani botanists ahead of the publication of the 3rd edition of the Red Book of the Republic of Azerbaijan, due to be published in 2023. Two further books have already been written - "Red Book of Nakhchivan Autonomous Republic" and "Rare plants of Ganja-Kazakh region". In order to preserve these studied species, information about the species is collected and action plans for in situ and ex situ conservation are prepared. It is especially important to study the rich flora and biodiversity of the Karabakh territory, characterised by its climate and landscape diversity. There are 121 rare Azerbaijan and Caucasus endemics found across Karabakh, which is famous for its unique flora but has remained unstudied for 30 years due to conflict. Most of these plants will need to be added to the 3rd edition of the “Red Book”.

A white flowered plant growing out of a bed of grey shale viewed from above next to a GPS device showing the altitude as 3981 feet
Cerastium latifolium L. (Caryophyllaceae) at an altitude of 3989 m above sea level, the Greater Caucasus Mountains. (Photo: Elman Yusifov).

The Institute of Botany and the Millennium Seed Bank Partnership have been cooperating bilaterally for 10 years to protect biodiversity. The main goals of the partnership are the implementation of joint new projects, the exchange of personnel and experience, and joint research on priority areas. So, as a result of currently implemented joint projects, the seeds of approximately 40% of the rare and endangered higher plants included in the 2nd edition of the Red Book of the Republic of Azerbaijan have been collected. These collections are stored in the Seed Bank of the Institute of Botany and duplicated at the Millennium Seed Bank and are important for the protection of those species in the future and for the implementation of restoration and reintroduction work over the coming years.

A group of 3 people crouching down around a plant specimen, one with a camera, one taking notes and one touching the plant. With mountainous hills and a barren, grassy landscape in the backgound
Field work in the upper mountain belt, Lerik district. (Photo: Aida Ibrahimova).
A dense sward of pink clover infloresences and leaves with one white clover in the centre. Amongst the clover are pink Geraniaceae flowers and their dissected leaves. Grass leaves are poking up through the sward
Highland meadow vegetation. (Photo: Sayyara Ibadullayeva).

Since its opening in 2006, the collections at Bali Botanic Gardens’ seedbank mainly came from garden plants (Lestari & Asih, 2015). Recognising the need to increase genetic diversity for restoration and re-introduction purposes (Rao et al., 2006), collecting wild plant seeds began on Nusa Penida in 2018, and then in Bali Barat National Park in 2019. The seedbank also started to receive seed collections from Bali Botanic Gardens’ flora exploration teams at that time.

However, seed harvest and collection management efforts at Bali Botanic Gardens’ seedbank still did not entirely comply with the standardised seed banking procedures recommended by Kew’s Millennium Seed Bank. Increasing the capacity of staff at Bali Botanic Gardens was seen as a way to address this condition which might otherwise hamper the seedbank’s conservation effort. The capacity improvement need was highlighted further by the fact that in 2016, Bali Botanic Gardens seedbank had become part of the collaboration between the Indonesian Botanical Gardens Seedbank and the Millennium Seed Bank Partnership (MSBP).

Staff from Bali Botanic Gardens attended several capacity building activities hosted by the MSBP Kew team. The first was the regional Seed Conservation Techniques course held in Cibodas Botanic Gardens from 25th - 29th April 2017 (Figure 1). In the same year, a member of staff from Bali Botanic Gardens attended the 19-day Seed Conservation Techniques training course held at the MSB, Kew. These courses were very beneficial as the material and practical activities were provided directly by the Millennium Seed Bank teams.

A row of people all stood smiling in front of a wooden building holding a banner reading Regional Seed Conservation Technique Course
Figure 1a. Regional seed conservation techniques course at Cibodas Botanic Garden. (Photo: BRIN).
A group of people stood around a table with plastic trays on it listening
Figure 1b. Regional seed conservation techniques course at Cibodas Botanic Garden. (Photo: BRIN).

Seed banking capacity improvements at Bali Botanic Gardens needed to reach as many people as possible. Transferring knowledge to one or two staff members would not have been practical for the long term as staff at the seedbank are continuously changing. Thus, in September 2020 Bali Botanic Gardens seed banking staff met with others from all Indonesian Botanic Gardens to share and discuss their experiences in seed collection following MSBP standards. As travelling at that time was restricted due to COVID-19, the meeting was held virtually. The meeting gave staff valuable insight into seed and data collection processes to be used during and after field trips. However, confusion and uncertainty remained in the minds of staff as most had never used these processes first-hand.

6 people sat around a desk a large projector screen on the wall with a virtual meeting happening.
Figure 2: Virtual meeting to familiarize Indonesian Botanic Gardens Seed Banking team with seed collection procedures. (Photo: Bali Botanic Garden Seed Banking Team).

As a follow up, two seed and data collection practice sessions were conducted in October 2020, with the objective to provide staff with a glimpse of what they would do during the seed collection process. Both sessions were held in Bali Botanic Gardens, first with the Proteaceae collection and then with the Legumes collection (Figure 3). Although the first session was pretty challenging, the second went more smoothly as the staff were already familiar with the procedure and forms that would be needed in the field.

Four people stand in the Bali Botanic Gardens, two look up to the tree canopy, one leans against a tree trunk reading some notes, the other stands looking at her clipboard
Field practice on seed collection. (Photo: Bali Botanic Garden Seed Banking team).

In 2021, nine Bali Botanic Gardens staff participated in a virtual seed banking training session held by the MSBP, in partnership with various stakeholders in Indonesia. For this training, a small number of staff participated together in the office, to support those unfamiliar with virtual meetings (Figure 4).

Three people sit in a classroom looking at a microsoft teams meeting projected on the wall
Bali Botanic Garden staff participate in a virtual seed banking course. (Photo: Bali Botanic Garden Seed Banking team).

The training was very beneficial and provided the staff with the basic knowledge needed to maintain collections in the seedbank, including how to conduct seed viability tests, collection data management and other aspects regarding seed collection maintenance. The training also provided an opportunity for the staff to interact with the MSBP team, discuss and clear all confusion regarding seed banking procedures. The training gave staff a much more comprehensive understanding of the seed collection process and know how to maintain their collection in the seedbank.

We hope that the capacity strengthening conducted over the years will improve the standard of the Bali Botanic Gardens’ seed banking efforts in the future. We also hope that by involving a wider group in the capacity building activities, including staff not currently posted at the seedbank and a university student (at the second field practice), we will ensure the longevity of the seed banking efforts.

References
  • Lestari, D. & Asih, N.P.S. (2015) Pengelolaan bank biji Kebun Raya Eka Karya Bali. Proc. Masy. Biodiv. Indonesia. 1(3): 515-520.
  • Rao, N.K., Hanson, J., Dulloo, M.E., Ghosh, K., Nowell, D. & Larinde, M. (2006) Manual of Seed Handling in Genebanks. Biodiversity International, Rome.

The Australian Black Summer of 2019-20 saw the western half of Kangaroo Island, South Australia burnt by intense bushfires. More than 167,000 hectares were affected, largely comprised of remnant bushland, with 83% of the area burnt at high to very high severity. Kangaroo Island, along with the adjacent Mt. Lofty Ranges, is one of Australia’s 15 ‘Biodiversity Hotspots’ – concentrated areas of high biodiversity characterised by elevated levels of endemism.

A landscape with black burnt shrub branches sticking up in stark contrast to the sandy soil. Some of the shrubs are showing evidence of regeneration with leaves sprouting from the base
Burnt landscape of Kangaroo Island post-fire. (Photo: SASCC).

The team from the South Australian Seed Conservation Centre (SASCC) arrived on the island shortly after these catastrophic fires to assess the post-fire response of some of the island’s endemic and threatened flora and, where possible, collect germplasm for long-term storage at the Botanic Gardens and State Herbarium, South Australia. Sensing the community’s desire to not only restore what was lost during the fires, but also to be better prepared for the next fire event, the SASCC devised the novel concept of a ‘seed production garden’.

A grant delivered through the Australian Seed Bank Partnership, funded by the Commonwealth Government of Australia, was used to develop the Kangaroo Island Seed Production Garden (SPG). The vision is to create an aesthetic garden of genetically diverse, living plant collections that can be used for conservation, restoration and translocation, population supplementation, research and seed banking, and is supported by the local community, benefactors (Paton Family), business (Troppo Architects) and several environmental NGOs (Bio-R, Nature Conservation Society of South Australia). An important aspect of this community-led garden will be to introduce members of the local community and visitors to the island’s threatened flora. Targeted seed production will also bridge an existing gap in the restoration of self-perpetuating populations of many threatened plant species on the island by ensuring the availability of sufficient, genetically robust and sustainably sourced propagules.

A photo montage of artists impressions of the Kangaroo Island Seed Production Garden from different angles, showing a raised open building with a slanted roof with solar panels. There is a small open seating area looking out over the gardens. The gardens have lots of different beds spread out with curved paths running between them
Concept design for Kangaroo Island Seed Production Garden. (Photo: Troppo Architects).

The development of the SPG is the result of almost two decades of seed collecting trips to the island by the SASCC team. The island boasts 60 endemic plant species, many of which were already germplasm-banked at the Botanic Gardens in Adelaide at the time of the fire. The SASCC was able to utilise some of this material – often collected when wild populations were larger and most likely had higher levels of genetic diversity – to propagate plants for introduction to the SPG. Excess propagules were also distributed to conservation-minded community members to augment existing populations or to establish new populations.

While the concept of seed production areas is nothing new, the idea of marrying utilitarian seed production with an inviting, aesthetic garden setting may be. Removed from the abstractions of formal reports and scientific literature, viewing a diverse collection of threatened plants up close and in a single convenient location makes the germplasm collection and propagation aspects of plant conservation tangible and hands-on. Rather than traversing dense vegetation to reach an isolated swamp in a remote Wilderness Protection Area at the western end of Kangaroo Island, a volunteer or visitor can witness the splendour of the Vulnerable, midday blooming aquatic Nymphoides geminata (Menyanthaceae), collect seeds from dehiscing fruits and propagate plants for translocation – all just 20 minutes from the island’s major township of Kingscote.

A yellow flower with 5 petals each with a fringed margin
Nymphoides germinata blooming. (Photo: SASCC).

The half-hectare garden has been fenced to exclude browsing kangaroos, wallabies and possums and is landscaped with different soils and water features to match the habitats of the species being grown. On 9th July 2022 the SPG was officially opened by the State Minister for Climate, Environment and Water, the Hon. Susan Close MP, in the company of dozens of community members. During the opening the first 1,500 plants comprising over 60 threatened and endemic species were planted into the landscaped beds by eager volunteers.

A large group of volunteers within a fenced enclosure all working to plant out seedlings from plant pots spaced out over mounds of soil
Volunteers at the Seed Production Garden opening day. (Photo: SASCC).

At present, only one quarter of the garden has been planted, with enormous scope for the inclusion of additional species, multiple provenances and in-ground germination research. The SASCC team is now working on harnessing the public interest that the development of the garden has received into a ‘Friends of the Kangaroo Island Seed Production Garden’. This group will have a large ongoing role in the operation of the garden. The SASCC will provide technical guidance, including training volunteers in seed collection techniques, plant propagation and searching for new wild populations of target species. After what was for many Kangaroo Island residents a traumatic event, it has been wonderful to see people’s energy focused towards what will hopefully be a long-lived and substantive contribution to plant conservation in South Australia.

During a visit by MSB coordinators to Indonesia in August this year, they interviewed Dr Dian Latifah about her experiences of how she got into seed banking, her job and what are the challenges for seed banking in Indonesia.

Dr Dian Latifah stood next to Liz Truss presenting her with a book. They are stood in front of a round memorial building of 8 large white pillars with a central plinth with a plaque on.
Dr Dian Latifah welcoming British Prime Minister Liz Truss, at the time Secretary of State for Foreign, Commonwealth and Development Affairs of the United Kingdom, to Bogor Botanic Gardens in 2021. (Photo: Dina Safainanugraha).

How did you become interested in plant conservation?

Since I was a kid I loved growing plants in our garden, and my parents loved it too. That influenced me to take my BSc in Agronomy at IPB university. I did my BSc thesis on the seed science of rice. Then I applied to LIPI to be a seed scientist working on wild plants. I’ve been doing research on seed conservation since then.

My MSc and PhD were also on seed science, this time on palm regeneration of wild palms in Queensland. I spent 5½ years in Australia.

How has seed conservation at Bogor Botanic Garden (BBG) changed since you began working?

When I started working at Bogor Botanic Gardens, they collected orthodox seeds from the gardens, didn’t measure their moisture content and stored them at room temperature. Our director at the time, Dr Didik Widyatmoko (now he is a research professor), wanted to improve seed bank procedures, so my colleagues the late Mr Sutrisno, Dr Siti Roosita and I wrote a proposal to get state funding for seed bank development. After we got the funding, I searched online to find where I could do training and discovered the Millennium Seed Bank (MSB). I contacted the MSB and attended the Seed Conservation Techniques course in 2015, and our partnership developed from there.

Why is seed conservation important to BRIN (Research Centre for Plant Conservation, Botanic Gardens and Forestry at National Research and Innovation Agency of the Republic of Indonesia) and Indonesia?

At the start, the strategy for ex situ conservation, consisted of living collections and seed banking. Both have pros and cons. E.g., Living collections need more space, but the seed bank only needs a little space, and we can store a much greater diversity of plants. Therefore, we focused on both. We follow GSPC Target 8 that 75% of a country’s biodiversity should be stored ex situ in-country, and seed banking contributes to that. From the start, we have been building the collections through field work, prioritising rare and endemic plants, and those with economic potential, for example, Myristica spp. (nutmeg) and Eusideroxylon zwagerii (ironwood).

A seed bank is a big investment, and we need to show value for money. We do this through increasing collaboration with students, academics and the public.

Indonesia is a very large country, with lots of islands. You have two big seed conservation projects with the MSB. How do you organise your teams to collect from the different parts of Indonesia?

At the start I asked for proposals from my colleagues at the four botanic gardens in Bogor, Cibodas and Purwodadi (Java) and Eka Karya (Bali) and submitted them to Kew. We now have 11 sub-projects covering different geographic regions, aiming to collect over 1,000 species, under the Arcadia Threatened Biodiversity Hotspots project. This project lasts for three years until 2024 and each year we organise field trips to different islands, from Sumatra to Papua. We chose several National Parks and Nature Reserves that have high biodiversity and where we have built up a good relationship with the authorities

What are the challenges for seed collecting in tropical forests?

(Laughs). First, we are dealing with remote areas and challenging weather – it rains a lot so it may limit the time available for seed collecting and reduce access to the areas. There is also a lack of information on fruiting seasons – we may get some advice from the park rangers, but when we arrive the fruits are immature. Then there is often a lack of information on seed storage behaviour on references like the Seed Compendium, other articles, SID and the Wyse predictor – sometimes we are not sure if a species is orthodox or not, and when we try to do storage or germination experiments it seems to be recalcitrant. It is difficult to get enough seeds, up to 10,000. On average, we often can only get from 250 to 3,000. We may only be able to find a small number of trees, often less than five, and the ones we can find may be inaccessible. Finally, we need up to seven different letters and permits with named target species to collect and conserve seeds from protected areas and it may take weeks to get them, so we have to start planning well in advance of each trip.

How would you like the Kew-BRIN partnership developing in the future?

We would like to extend the project in Mt Ciremai National Park, to achieve more significant results. The current project, funded by the Garfield Weston Foundation, is a combination of seed conservation and utilisation for restoration, that involves several parties, including Mt Ciremai National Park, Kuningan Botanic Gardens / Kuningan District Council, BRIN and Kew. The National Park has an ecosystem restoration programme underway, so our project will support this. We can involve Kuningan Botanical Garden as they have nursery skills. BRIN and KEW bring additional technical and scientific resources and help provide training and develop facilities.

The Arcadia project will end in 2024 and we would like to do more research on the seeds we have collected and monitor their viability, to learn more about the seed storage behaviour and longevity. For example, we have found problems with conservation of Begonia seeds, many of them are endemic and also have economic potential as amenity species. We’d also like to do more projects, such as setting up seed hubs in other parts of Indonesia for local seed storage and use.

A row of people all wearing facemasks stood against a wall. Above them on the wall is a sign which reads LIPI Bank BIJI Seed Bank
Dr Dian Latifah together with Head of Research Centre for Plant Conservation, Botanic Gardens and Forestry as well as colleagues from Bogor, Cibodas and RBG Kew as part of an MSBP Standards Review in August 2022. Picture taken at Indonesia's national seed bank at Bogor. (Photo: Dr Dian Latifah).

What is the most interesting plant you're working on at the moment?

Eusideroxylon zwagerii (ironwood) is an important species as it’s endangered and its timber is very high quality and has great economic value. It is known to have recalcitrant seeds, so we have been doing research on its seed storage behaviour and the best method of short-term storage. We can now store its seeds for up to three years!

Mouldy versus infested seeds? Scoring X-rays and germination tests

Can an X-ray detect fungal, bacterial or viral infections in a seed? When should seeds be classified as 'mouldy' or 'infested' in a germination test? Do insect infestations differ from fungal, viral or bacterial infections when scoring germination tests? We asked Sarah, Nicola and Rachael from the Millennium Seed Bank (MSB) Seed Collections Team to talk us through the differences.

Can you spot seeds infested with fungus on an X-ray? And how should these be scored?

At the seed quality checking stage of seed processing, a seed counted as 'infested' can relate to either insect damage/ presence or a bacterial/ fungal/ viral infection.

Whilst an X-ray cannot always tell us if a seed is infected with a fungus, bacteria or a virus, sometimes it can be seen for example, as a darker grey 'shadow', spreading out across the seed from one point. This often has a vaguely granular texture rather than the pure white of a healthy seed. A seed can also be counted as 'infested' at this stage if the infection is so severe it is clear that the seed is non-viable from the outside. Or for some smaller seeded species, for example with many Scrophulariaceae, the contents can still appear solid on an X-ray. In these cases, you can do a cut-test instead of an X-ray to determine the internal morphology and quality.

The signs of insect infestation to look for on X-rays are tunnels, usually of increasing size, with a cavity at the end where the lava may be (Figure 1a). In some cases, a seed may have been almost entirely eaten by an insect, in which case the seed will appear as almost empty with a few lighter spots of content (likely frass) left behind (Figure 1b). Insect infested seeds also sometimes visibly show exit holes for larva on the outside of the seed coat, for example with some Fabaceae species. However, it should be noted that insect infestation does not necessarily mean that the seed is inviable, it can depend on how much of the seed has been consumed. At the MSB there is a ‘part full’ category alongside ‘full’, ‘empty’ and ‘infested’ for scoring X-rays which can be used if there is some uncertainty as to whether enough of the seed embryo remains (has not been eaten) for the seed to still be viable.

Many white oval healthy seeds, but three seeds are highlighted by red circles. The oval embryos of these seed are greyer than the other and each contain a small dark curved arc within them where the larva is
Figure 1a: An X-ray image of the seeds of Fraxinus excelsior showing larval infestation in some of the seeds (red circles). (Photo: RBG Kew).
Lots of white healthy oval embryos and a few which are duller grey indicating empty seeds. Two seeds are circled red, these seeds are mostly dark grey bust have smaller lighter patched at one end
Figure 1b: An X-ray image of the seeds of Fraxinus excelsior. Red circles highlight examples where the majority of the contents has been eaten by insects and minimal contents remain. (Photo: RBG Kew).

Should those seeds which are black at the end of the germination test be recorded as 'mouldy' or 'infested'?

NB. When you are setting up a germination test, if you have any concerns about the seeds, you can do a cut-test on a sub-sample prior to setting up the test.

At the MSB we use six different categories to classify seeds during a germination test (Table 1). But when you’re scoring your test as you go along or are doing your cut-test at the end, you may have some degree of fungal growth and it is not always clear under which category the seeds should be classified.

Table 1: Germination test seed classifications
Category Include in germination calculation?
Germinated Included as 'germinated'
Abnormal Included as 'not germinated'
Fresh Included as 'not germinated'
Mouldy Included as 'not germinated'
Empty Not included
Infested Not included

'Infested' varies from 'mouldy' in that a seed should be classified as 'mouldy' if you can see signs that there was suitable initial morphology at the start to enable germination, i.e., an embryo was present (Figure 2a). If this is not possible to detect the seed should be classified as 'infested' (Figure 2b). If a seed is classified as 'mouldy' it could suggest that the seeds have either lost their viability over time, there was poor test hygiene, or that the germination conditions were incorrect, e.g., too warm.

The two halves of the seed are deformed and light brown in colour, there is a dark creamy colour to the inside of the seed but it does not look solid
Figure 2a: A cross section of a mouldy Daucus carota seed. (Photo: RBG Kew).
The two halves of the dissected seed each have a small hollow within them where the embryo within the seed has been eaten. At the ends of the seed are small sections of the embryo remaining but not enough for germination to occur
Figure 2b: A cross section of an infested Daucus carota seed. (Photo: RBG Kew).
A cross section of a seed showing a clearly defined light brown seed coat and a creamy white embryo with no sign of damage.
Figure 2c: A cross section of a viable Daucus carota seed. (Photo: RBG Kew).

Is it possible for fungal spores to spread from one seed to another during storage?

It would be possible for a fungus to spread between seeds during the cleaning process if equipment is not cleaned properly between uses or if the seeds come into close contact with an infected batch whilst drying. Once the seeds are dried and sealed for banking, assuming they are fully sealed, it is less likely for fungal spores to transfer between collections. The other possible source of fungal spores for infection could be the medium used for germination testing, e.g., agar, or from the air whilst setting up the test. If you suspect seeds could be infected with fungal spores, it is possible to sanitise the seeds prior to starting the germination test. This can be done by soaking the seeds in 0.5% sodium hypochlorite (NaOCl) solution, containing a 1% surfactant (Tween 20) for 10 minutes and then rinsing under running water for 1 minute (see Technical Information Note 13a). Once the germination tests are set up, the container they are in should be sealed inside a plastic bag to reduce the chance on contamination (this also helps to prevent moisture loss). In addition, 'mouldy' and 'infested' seeds should be removed from the test as it progresses to limit the influence they have, and tests can be re-plated during the test if fungal growth gets too bad.

Further information on setting up germination tests can be found in Technical Information Note 13a.

Recent publications from across the MSBP:




Issue 3: June 2022

In this issue we hear about field adventures in Madagascar and Zambia, research studies from Georgia, Italy and the UK, and about the latest edition of the Australian Plant Germplasm Conservation guidelines. We interview Naomi Carvey, MSBP Data Warehouse Officer at the MSB and the species profile is the US endemic Daucosma laciniata.

Leaves and small round green fruits Psydrax sambiranesis
Psydrax sambiranensis, Madagascar. (Photo: Vonona Randrianasolo, KMCC).
A pot with a single seedling growing showing the leaves with pairs of leaflets
Daniellia alsteeniana seedling, Zambia. (Photo: David Mwale, FDR).
Close up of Daucosma laciniata in flower, with the white flowers within the umbel and the divided bracts.
Daucosma laciniata, USA. (Photo: Bill Carr, Lady Bird Johnson Wildflower Center).
Five plant pots each with a different species of Brassica being grown showing a variety of leaf morphologies
Brassica seedings, growing at the MSB. (Photo: Pablo Gómez).

Madagascar is committed to achieving Target 8 of the Global Strategy for Plant Conservation, and in the spirit of any post-2020 targets, we wish to ensure that all our threatened flora is conserved, and available as ex situ material for future conservation, restoration and research. For MSBP Madagascar this means, we want to rapidly increase the number of threatened species conserved as seed collections, where possible. The Kew MSBP Madagascar team, in collaboration with our national partners, the Silo National des Graines Forestières (SNGF) is implementing the Global Tree Seed Bank Programme and we have adopted a new collection strategy based on geographical gaps in our current collections. An analysis of the collection density over the years has given us a clear picture that the majority of the collections made so far, have been from outside protected areas. To compensate for this spatial anomaly, field trips carried out during 2022 are focussed on protected areas, in order to cover as many sites as possible to ensure the collection and ex situ conservation of species of restricted distributions, including large numbers of more threatened species.

Initially, six potential sites located in the northern part of Madagascar were chosen, four of which were new to the MSBP. These are the Kasijy, Ankarana, Andrafiamena Andavakoera and Analamerana Protected Areas. Each site has its own climatic and geographical specificities that affect the distribution of the different types of vegetation. There is xerophytic vegetation on the Tsingy (Ankarana, Montagnes des Français Protected Areas), deciduous dry forests (Kasijy, Analamerana, Andrafiamena Andavakoera, Ankarana Protected Areas) and degraded dry forest in the rocky hills around Ambilobe.

A limestone outcrop with trees and shrubs growing on it and trees in the foreground.
A vegetated limestone outcrop in Montagne des Français. (Photo: Vonona Randrianasolo).

In Madagascar, accessibility to many sites by vehicle is almost impossible especially during the rainy season, and this has been made worse as a result of the cyclones we have witnessed over the past few years. The team's fieldwork in January and March coincided with the passage of two cyclones in the north eastern part of the island, which subsequently led to heavy rains in the region. This has greatly affected our fieldwork plans. For example, the visit to Andrafiamena Andavakoera had to be abandoned due to the rise of the Irodo rivers that have to be crossed to reach the site. Our Landrovers are good, but 6ft deep rivers in spate are a match for any wading vehicle!

A muddy pathway leading to shrubland with two seed collectors carrying collections and equipment
Access path to Analamerana Forest. (Photo: Nomentsoa Randriamamonjy, KMCC).
Two men in a dug-out canoe crossing a wide river
Crossing the Irodo river in Analamerana Protected Area. (Photo: Nomentsoa Randriamamonjy, KMCC).

The team had a memorable 14-day collection trip recently to Kasijy. From our base it is a two day drive to the regional headquarters where we spent the next day securing access and collecting permits. We then had a two and a half day trek simply to reach the forest edge. The subsequent four days spent collecting yielded 14 collections, all new for us. I am very proud of the team commitment this effort showed.

Working in new sites is a big challenge especially with a poorly known flora like Madagascar. A lot of the information we have for our target species does not correspond with what we find on the ground. Predicting fruiting periods and seed maturity based on herbarium data has not been very successful and for many sites, we do not have the network of collaborators on the ground to monitor phenology. We are also witnessing a wide variation in periods of seed maturity between species at a particular site. Securing seed from many species on one site visit is therefore, a challenge. Many of these protected areas are void of roads and paths and it has proven very difficult to assess the total population size for our target species.

Of the 82 collections made during these six collection missions, 11 are thought to be new to the MSB collections and 19 are threatened species.

Among the many interesting trees harvested during these field trips was Psydrax sambiranensis. Assessed as Critically Endangered (CR) and known only at one site in the rocky hills around Ambilobe district, it is threatened by rock mining for construction. With the post harvest work of SNGF, all these seed collections have now been cleaned and are ready for duplication to the MSB for long-term storage.

Leaves and small round green fruits of Psydrax sambiranesis
Psydrax sambiranensis (Critically Endangered). (Photo: Vonona Randrianasolo).
Four people sitting on a plastic sheet in a grassy area, cleaning a pile of seeds
Cleaning seeds in the field. (Photo: Ando Andriamanohera).

Daniellia alsteeniana P.A.Duvign. locally known as Mukulabushiku in our Bemba language is a conspicuous and very ornamental tree with large pink flowers growing to some 25m. It belongs to the Caesalpinioideae sub-family of legumes, and in the Northern parts of Zambia it is considered a threatened species through over-exploitation for its timber and resin. It is highly valued by local people providing construction materials for housing, canoes, crates and decorative veneer.

Currently the species is Near Threatened on the IUCN Red List and whilst we recognise that large, healthy populations of the species are still to be found in Angola and the DRC, due to its over-exploitation in Zambia we made this a priority for our collection programme. We intend to make seed material much more widely available for the many restoration projects currently being launched amongst these Miombo woodlands in Zambia. Lab-based germination reaches 98% and seeds are easily raised in field nurseries.

Four members of the FDR/MSB team stood next to a bench. The bench has trays of seeds on and equipment used for seed processing including funnels and jars.
The FDR/MSB team processing seed of D. alsteeniana. (Photo: FDR).
A pot with a single seedling growing showing the leaves with pairs of leaflets.
D. alsteeniana seedling raised from seed collected under FDR/MSB partnership. (Photo: David Mwale, FDR).

Herbarium vouchers stored at one of the largest and oldest Zambian National Herbariums (NDO) located in Kitwe, show plant specimens collected in Kasama (1957) and Kawambwa (1958) in Northern and Luapula provinces of Zambia, respectively. These 70-year-old records suggested fruiting occurred between October and November but following our collection trips to these areas in 2019, we found the seed was actually at the point of natural dispersal during August and September. Was this change in fruiting time something to do with our changing climate or simply a response to variation in the time of the rainy seasons? Our collection will add to our understanding of the fruiting behaviour of this species. We now have a good herbarium voucher, useful field images and a modest seed collection of some 3000 seeds stored here in Zambia with a safety duplicate at the MSB. We intend to revisit these provinces in the coming seasons to bulk-up these numbers with new collections.

The species dispersal mechanism is interesting. Each fruit (pod) contains 1-2 bi-convex oval and very glossy brown seeds each weighing about 4g. The funicle is persistent and so it seems to be the detached valve of the pod that falls, like a whirling samara, thus assisting dispersal of the attached seeds beyond the shadow of the crown.

A tear drop shaped seed pod split in half, at the pointed end of each half a brown  oval seed is attached via the funicle.
The dehisced seed pod showing the seeds attached at the point of dispersal. (Photo: Mike Bingham).
Five oval brown seeds arranged in a circle each with a long grey persistent funicle pointing into the centre.
Showing the large seed size with the persistent funicles. (Photo: David Mwale, FDR, Zambia).

Modern plant taxonomy considers blackthorn (P. spinosa) as a conservative species. Fundamentally, phylogenetic niche conservatism refers to the tendency of species to retain their ancestral traits. There is a substantial difference in the interpretation of this term by Russian and European schools of botany. The floras of the USSR and post-soviet countries are rich in plant species with morphological, and in some cases, even phenological characteristics which vary over short distances across their spatial range.

The floras and plant nomenclatural checklists of the Black Sea countries of Ukraine, Moldova, and Turkey suggest the existence of blackthorn species, subspecies, and variations that share nearly the same ancestral traits as P. spinosa. These are: P. podolica Andrz. ex Trautv.; P. spinosa var. dasyphylla (Schur) Domin; P. kurdica Fenzl ex Fristch; P. spinosa subsp. dasyphylla (Schur) Domin; P. stepposa Kotov. Most of them, except for P. spinosa subsp. dasyphylla, are synonyms of P. spinosa according to the Global Biodiversity Information Facility (GBIF). The old edition of the flora of Georgia (Sosnovsky, 1949; Gulisashvili, 1965) and the modern updates of the plant nomenclatural checklists for the country (Gagnidze et al., 2005; Davlianidze et al., 2018) indicate the distribution of the single taxon P. spinosa in the country. However, the local botanist N. N. Bregadze discovered five sub-taxa (all considered synonyms in GBIF) within the local population of this species:

  • var. ovaliputaminata,
  • var. subglobosoputaminata,
  • f. globosodrupacea,
  • f. ovalidrupacea,
  • f. plano-subgloboso-drupacea.

The description of the morphological characteristics of these varieties and forms is provided in the study “The Botanical and systematical research of the diversity of Prunus spinosa L.” (Botaniko-sistematicheskoe Izuchenie Prunus spinosa L. in Russian) published in 1976. The study was soon followed by a series of more comprehensive studies on the diversity of the Prunus genus L. in the South Caucasus conducted by another local botanist Eugenia Baiashvili who revealed the existence of differences in the chromosome number of individuals of P. spinosa in the country. The compilation of her articles on this topic was published as a monograph in 2010 (Baiashvili, 2010).

The discovery of the higher level of intraspecific variability inspired us to conduct a case study where we researched the population diversity of blackthorn in Georgia with the use of the modern methods of morphometry and plant genetics. The research was carried out as part of the “Enhancing Rural Caucasian Community Livelihoods Through Fruit and Nut Conservation” project and responded to its general aim of collecting and interpreting information about the globally and locally threatened species to raise local public awareness about the diversity of these key groups of plants.

The morphological study revealed three clusters of individuals, two clusters (clusters A, B; Fig. 1) weakly separated individuals of P. spinosa on the basis of the height of the shrub and leaf size i.e., these clusters included large and small-sized shrubs which are not associated with specific habitats or geographic sites in Georgia with their distribution. The third cluster (Cluster C, Fig. 1) strongly separated morphologically distinct forms with oval-shaped leaves and elliptic fruits and fruit stones - generally distributed in drylands of the Samtskhe-Javakheti region in the south part of Georgia (Fig.'s 1 & 4).

Graph of PCA results along two axes, PC1 = 45.75%, PC2 = 25.8%
Figure 1: A graph showing the results of the Ordination analysis with three different clusters shown in red, black and blue symbols. A text description of this figure is available in the body of the text.
Outline map of Georgia
Figure 2: A country map of Georgia in green with the distribution of P. spinosa collections shown in red dots. A text description of this figure is available in the body of the text.

The results of the genetic study showed weak divergence both on the intra and interspecific levels in the studied samples which supports the assumption that the species emerged after quick evolutionary radiation in the plum genus. Despite the low resolution of the genetic divergence of the taxa in the studied groups, the results supported the data of the morphometric study by proving the existence of the difference in haplotype structure between the group of individuals of P. spinosa distributed in the south part of Georgia and the rest of the individuals of this species collected in the other parts of the country.

In the foreground, a line of P. spinosa shrubs leading to a researcher standing in the distance next to the shrubs.
Figure 3: Large shrub form of P. spinosa. (Photo: Ana Kvlividze).
A researcher sitting next to P spinosa shrubs and studying leaves whilst holding a notebook and pen.
Figure 4: Small shrub form of P. spinosa. (Photo: Tolkha Shetekauri).
In the foreground a P.spinosa shrub next to a stone outcrop with other trees and a blue sky in the background.
Figure 5: Morphological form of P. spinosa with oval-shaped leaves and elliptic fruits distributed in South part of Georgia. (Photo: Ana Kvlividze).

Our results support the observation of the local scientists N. N. Bregadze and E. Baiashvili, and provide a more complete understanding of the diversity of blackthorn and its distribution in the country. We plan to continue the research in the future to discover the diversity of the plum genus at larger geographical scales in the Caucasus. With the taxonomical and ecological aspects of the research, it is important to focus also on the collection of ethnobotanical information. Fruits of blackthorn are collected in all parts of the Caucasus by local people because of their gastronomical and medicinal properties. They are even sold in the small countryside and large markets in Georgia. In this respect, it will be interesting to find out if local people differentiate blackthorn fruits by taste or any other property as this information would serve as additional proof of our findings on the existence of morphological and genetic differences between the sub-populations of P. spinosa.

Acknowledgements

The research was carried out within the framework of the project “Enhancing Rural Caucasian Community Livelihoods Through Fruit and Nut Conservation” funded by the Darwin Initiative Foundation in collaboration with the National Botanical Garden of Georgia, the Botanical Institute of Ilia State University, Georgia, and Royal Botanic Gardens, Kew from 2017-2021. As a result of this study, an MSc thesis with a similar title to this research was defended by A. Kvlividze at the Ilia State University in Georgia on February 18, 2022.

We are grateful to the project managers: Aisyah Faruk, Ian Willey, and David Kikodze, and Dr. Tsira Mikatadze-Pantsulaia - Head of the Plant Conservation Department at the National Botanical Garden of Georgia, for supporting this research.

References
  • Baiashvili, E. (2010) Phylogenetic study of genus Prunus L. (‘Kliavis (Prunus L.) Gvaris Philogenetikuri Shestsavla). Ed. Akhalkatsi, M. ; Botanical garden and the Institute of Botany. Tbilisi, Publishing house ‘Universal’. 130 pp.
  • Davlianidze M., Gviniashvili Ts., Mukbaniani M., JinjoliaImnadze L., Jugheli, T. (2018) Nomenclatural checklist of the flora of Georgia (Sakartvelos Phloris Nomenklaturuli Nuskha). Tbilisi, Publishing house ‘Universal’. 298pp. (in Georgian).
  • Gagnidze R.I., Maier G., Nakhutsrishvili G.S. (2005) Vascular Plants of Georgia: A Nomenclatural Checklist. Tbilisi, Georgian Academy of Sciences. 247pp.
  • Gulisashvili, S. [Ed.] (1965). Flora of the woody plants of the Caucasus, vol.: 4 (Rosacea-Leguminosae)(Kavkasiis Dendrophlora, Tomi IV (Rosacea-Leguminosae)). The Academy of Sciences of Georgian USSR, Institute of Forestry. Tbilisi, Publishing house ‘Metsniereba’. 405 pp. (in Georgian)
  • Sosnovsky, D. (1949) The Flora of Georgia, vol.: 5 (Rosacea-Leguminosae) (Sakartvelos Phlora, Tomi: V (Rosacea-Leguminosae)). Ed. Kharadze, A. Publishing and press of the Academy of Sciences of Georgian USSR, Tbilisi. 134 pp. (in Georgian)

Limited availability of data relating to regeneration traits has curbed macroecological research into the germination niche (Saatkamp et al., 2019). However, seed banks that focus on the ex situ conservation of wild species routinely carry out research to identify seed dormancy and germination requirements (Hay & Probert, 2013), which have become a valuable source for large-scale studies into seed germination (Sentinella et al., 2020).

In Europe, the European Native Seed Conservation Network, known as the ENSCONET Consortium, was established in August 2010 with the aim of maintaining significant levels of ex situ seed conservation activity for European native plant species. It builds on the success of the ENSCONET project, funded under the European Commission’s Sixth Framework Programme for Research and Technological Development (FP6) between 2004 and 2009 (Eastwood & Müller, 2012). The objective of the initial project was to improve quality, coordination and integration of European seed conservation practice, policy and research for native plant species and to assist EU conservation policy and its obligations to the Convention on Biological Diversity and its Global Strategy for Plant Conservation (Müller, Eastwood & Linington, 2012). A total of 33 institutes across Europe currently constitute the ENSCONET Consortium, all of which have an explicit interest in seed conservation of European native plants. One of the successful outputs has been the establishment of the ENSCONET database (ENSCOBASE), an online database that has been storing data on European native seed accessions since 2005, covering all European geographical regions and major plant families (Rivière et al., 2018).

ENSCOBASE contains publicly available primary records of laboratory germination experiments from multiple seed banks across all European Biogeographical Regions. It holds information on experimental germination conditions (germination temperature, diurnal temperature regime, light and dormancy-breaking treatments), alongside seed viability records and the associated seed lot metadata (e.g. biogeographical region; Fig. 1). As such, ENSCOBASE represents a valuable source of information to understand the macroecology of seed germination. Yet, there are challenges to the analysis and interpretation of the data, mainly due to the inherent heterogeneity related to each seed bank employing differing seed processing protocols. This between-study variation, in terms of meta-analysis theory can be addressed by applying recent developments in phylogenetic meta-analytical methods (Garamszegi, 2014). Based on this approach, we recently conducted a meta-analysis of the germination data stored in ENSCOBASE (Carta et al., 2022), finding that climate, specifically drought and cold seasons, shape the germination niche of temperate plants. The methodology used was able to account for between-study variation relating to seed sources and physiological status, thereby highlighting how large datasets generated by conservation seed banking can be a valuable resource in addressing questions in plant macroecology and evolution. We started the process in 2018, and although initially we found the heterogeneity of the original data difficult to manage, the resulting experience and eventual outcome proved to be a worthwhile and impactful study of the European flora.

Outline map of Europe with different colours indicating the area covered by different biogeographical regions
Figure 1: Map of European Biogeographical Regions. A text description of the map is available. (Source: EEA).
References
  • Carta, A., Fernández-Pascual, E., Gioria, M., Müller, J.V., Rivière, S., Rosbakh, S., Saatkamp A., Vandelook F. & Mattana, E. (2022) Climate shapes the seed germination niche of temperate flowering plants: a meta-analysis of European seed conservation data. Annals of Botany. mcac037. DOI: https://doi.org/10.1093/aob/mcac037
  • Eastwood, R. & Müller, J.V. (2012) Achievements of the European Native Seed Conservation Network – ENSCONET. In: Maxted, N., Ehsan Dulloo, M., Ford-Lloyd, B.V., Frese, L., Iriondo, J., Pinheiro de Carvalho, M.A.A. Eds. Agrobiodiversity conservation: securing the diversity of crop wild relatives and landraces. pp. 286-291. Wallingford. CABI.
  • Garamszegi, L.Z. (2014) Modern phylogenetic comparative methods and their application in evolutionary biology: concepts and practice. Heidelberg; London. Springer Verlag.
  • Hay, F.R. & Probert, R.J. (2013) Advances in seed conservation of wild plant species: a review of recent research. Conservation Physiology. 1(1): cot030. DOI: https://doi.org/10.1093/conphys/cot030
  • Müller, J.V., Eastwood, R. & Linington, S. (2012) ENSCONET: a milestone for European seed conservation. Studi Trentini de Scienze Naturali. 90: 209–210.
  • Rivière, S., Breman, E., Kiehn, M., Carta, A. & Müller, J.V. (2018) How to meet the 2020 GSPC target 8 in Europe: priority-setting for seed banking of native threatened plants. Biodiversity and Conservation. 27: 1873-1890. DOI: https://doi.org/10.1007/s10531-018-1513-2
  • Saatkamp, A., Cochrane, A., Commander, L., Guja, L.K., Jimenez-Alfaro, B., Larson, J., Nicotra, A., Poschlod, P., Silveira, F.A.O., Cross, A.T., Dalziell, E.L., Dickie, J., Erickson, T.E., Fidelis, A., Fuchs, A., Golos, P.J., Hope, M., Lewandrowski, W., Merrit, D.J., Miller, B.P., Miller, R.G., Offord, C.A., Ooi, M.K.J., Satyanti, A., Sommerville, K.D., Tangney, R., Tomlinson, S., Turner, S. & Walck, J. et al. (2019) A research agenda for seed-trait functional ecology. New Phytologist. 221(4): 1764-1775. DOI: https://doi.org/10.1111/nph.15502
  • Sentinella, A.T., Warton, D.I., Sherwin, W.B., Offord, C.A. & Moles, A.T. (2020) Tropical plants do not have narrower temperature tolerances, but are more at risk from warming because they are close to their upper thermal limits. Global Ecology and Biogeography. 29(8): 1387-1398. DOI: https://doi.org/10.1111/geb.13117

Text description of Figure 1: Map of European Biogeographical Regions

A map of Europe showing the areas covered by 11 different biogeographical regions. The Alpine biogeographic region covers most of Norway, and is also present in the Pyrenees, the Alps, the Carpathian Mountains, the Balkan Mountains, the Caucasus Mountains, the Ural Mountains and a small section of the Apennines in central Italy. The Anatolian biogeographic region covers the central area of Turkey. The Arctic biogeographic region covers Iceland, Svalbard, Novaya Zemlya and northern Russia. The Black Sea biogeographic region covers extends around the coast of the Black Sea from Bulgaria, through Turkey and into Georgia (west to east). The Atlantic biogeographic region covers the United Kingdom, Ireland, northern Spain, westerns parts of France and Denmark, Belgium, the Netherlands and northern Germany. The Boreal biogeographic region extends from Sweden east covering Finland, Lithuania, Latvia, Estonia, northern Belarus and western Russia. The Continental biogeographic region extends from western France through southern and eastern Germany, Czechia, Poland, southern Belarus and northern Ukraine into Russia, also extending south into northern Moldova, eastern Romania and Bulgaria, Serbia, Bosnia and Herzegovina, northern Croatia and north-west Italy. The Mediterranean biogeographic region extends from Portugal and Spain, east into southern France, western areas of Italy and coastal areas of Croatia, Bosnia and Herzegovina, Montenegro, Albania, Greece and Turkey. The Pannonian biogeographic region covers Hungary, north into Slovakia and south into northern Serbia. The Steppic biogeographic region extends from western Romania along the Black Sea coast into southern Ukraine and Russia

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The third edition of ‘Plant Germplasm Conservation in Australia – strategies and guidelines for developing, managing and utilising ex situ collections’ was recently released by the Australian Network for Plant Conservation (ANPC) and the Australian Seed Bank Partnership (ASBP) (Fig. 1). These ‘Germplasm Guidelines’ are practical, technical and evidence-based, providing a workflow to address each step of acquiring, maintaining and utilising genetically representative collections (Fig. 2).

Plant Germplasm Conservation in Australia, strategies and guidelines for developing, managing and utilising ex situ collections in Australia. Third edition. Edited by Amelia J. Martyn Yenson, Catherine A. Offord, Patricia F. Meagher, Tony Auld, David Bush, David J. Coates, Lucy E. Commander, Lydia Guja, Sally L. Norton, R.O. (Bob) Makinson, Rebecca Stanley, Neville Walsh, Damian Wrigley, Linda Broadhurst.
Figure 1: Cover image of ‘Plant Germplasm Conservation in Australia’ 3rd edition. (Image: ANPC).
Six steps for deciding ex situ conservation options
Figure 2: Decision flowchart for ex situ conservation options. A text description of the flow chart is available. (Source: Offord, C.A., Makinson, R.O., Guja, L., Auld, T.D. (2021) Options, major considerations and preparation for plant germplasm conservation. In Martyn Yenson, A.J., Offord, C.A., Meagher, P.F., Auld, T.D., Bush, D., Coates, D.J., Commander, L.E., Guja, L.K., Norton, S.L., Makinson, R.O., Stanley, R., Walsh, N., Wrigley, D. & Broadhurst, L. (Eds.) Plant Germplasm Conservation in Australia: strategies and guidelines for developing, managing and utilising ex situ collections. Third Edition. Canberra. Australian Network for Plant Conservation).

These Guidelines bring together decades of research and experience in conserving Australian plants in seed banks, botanic gardens and conservation nurseries (Fig. 3). They take readers through the genetics and practice of acquiring collections and the processes of seed banking, tissue culture, cryopreservation, and living collections maintenance, with 50 case studies to highlight the application of research and theory. In this edition, we add new chapters on the role of the nursery and identifying and conserving non-orthodox seeds. We also address the need to capture and utilise ‘special’ types of germplasm (material from ferns, mosses, liverworts), or taxa with ‘special’ life history stages or growing requirements (terrestrial orchids with mycorrhizal associations, carnivorous and parasitic plants). With an eye to the future, we finish with new chapters on risk management, and collection maintenance and utilisation. We gratefully acknowledge the input of 78 contributors from across the restoration and agriculture sectors, botanic gardens, the Commonwealth Scientific and Industrial Research Organisation (CSIRO) and universities, as well as the Kew Millennium Seed Bank, Botanic Gardens Conservation International, the Center for Plant Conservation and the Exceptional Plants Conservation Network to update this edition; and thank The Ian Potter Foundation for funding this project.

A map of Australia split into the different states indicating the location and facilities of the major Australian flora conservation centres.
Figure 3: Location of major ex situ conservation facilities for Australian flora, including ASBP Partners, the Australian Tree Seed Centre, the Australian Grains Genebank and Australian Pastures Genebank (both storing crop wild relatives) and major forestry seed banks with conservation collections. A text description of the map is available. (Image: CAM Graphics; Offord, C.A., Makinson, R.O., Guja, L., Auld, T.D. (2021) Options, major considerations and preparation for plant germplasm conservation. In Martyn Yenson, A.J., Offord, C.A., Meagher, P.F., Auld, T.D., Bush, D., Coates, D.J., Commander, L.E., Guja, L.K., Norton, S.L., Makinson, R.O., Stanley, R., Walsh, N., Wrigley, D. & Broadhurst, L. (Eds.) Plant Germplasm Conservation in Australia: strategies and guidelines for developing, managing and utilising ex situ collections. Third Edition. Canberra, Australian Network for Plant Conservation.).

The Germplasm Guidelines were launched by Prof. Tim Entwisle during the Australasian Seed Science Conference in September 2021. This virtual meeting tackled issues across seed biology and ecology through to the sourcing, end-use and management of seed collections, as well as the roles that seeds play in local and global culture. As a fully-virtual meeting the conference attracted 425 delegates from 34 countries, enabling researchers to share their work, including the new edition of the Germplasm Guidelines with an engaged international audience. Additional funding from the Australian Academy of Science is enabling the ANPC to continue these discussions through the Australian Academy of Science Fenner Conference on the Environment: ‘Exceptional times, exceptional plants’.

Exceptional plants include those species for which seeds are not available (not produced at all, inadequately produced, or non-viable), species with non-orthodox seeds (desiccation sensitive, freezing-sensitive or short-lived), and species with deeply dormant seeds. While the work of seed banks is important, and although the majority of seed-bearing plant species are considered to be orthodox, we still face challenges with many species when it comes to determining how to store them longer-term in ex situ conservation facilities and regenerate them from seed.

Many conventional seed banks are already working on exceptional species in collaboration with botanic gardens, nurseries and other experts to identify alternative options for conserving exceptional plant species. The 3rd edition of Germplasm Guidelines presents examples from around Australia through several informative case studies and workflows which may be of assistance to facilities that have recently started or are planning to commence work on exceptional species.

The Germplasm Guidelines are available for free download or ordering of print copies from the ANPC plant germplasm webpage.

The written guidelines are supported by high-quality video content accessible via the 'Plant Germplasm Conservation in Australia' playlist on the ANPC YouTube channel.

Dr. Amelia Martyn Yenson stood behind a bench in a seed laboratory looking at a book
Project Manager and lead editor for the Germplasm Guidelines revision, Dr. Amelia Martyn Yenson at the Australian PlantBank. (Photo: Michael Lawrence-Taylor).

Text description for Figure 2: Decision flowchart for ex situ conservation options

A flowchart for deciding ex situ conservation options, referencing the relevant chapter and section in the Germplasm Guidelines.

The taxon or population is considered at risk or in need of ex situ conservation (Chapter 1) leading to the questions (Section 2.3):

  • Have all major considerations been addressed?
    • Is there a clear link to an in situ recovery process?
    • Is it a precautionary collection?
    • Is it for research or display?
    • Is it an ‘exceptional species’? (Chapter 6)
  • What is the time frame?
  • What information is available?
  • Is seed available (preferred in most cases)?
  • Will you need to conduct preliminary trials on collection, storage or propagation?
  • Do you have the necessary resources? Can you get the necessary permits or permissions?

Using these it is then asked do the benefits of ex situ conservation outweigh the risks (Section 2.2)? If no, monitor populations or pursue other options, if yes decide ex situ options. If seed is not suitable, consider other options (Section 2.4, Chapter 8). Then collect material appropriate to the options chosen (Chapters 3 and 4).

Collected material can then be used in four different ways:

  1. Seed banking (Chapters 5, 6 and 7)
  2. Tissue culture, orchid and symbiont culture (Chapters 9 and 12)
  3. Cryopreservation (Chapters 10 and 13)
  4. Living plant collections (Chapter 11)

Material from these four options can then be utilised (for translocation, research etc.) and appropriate risk management implemented (Chapters 14 and 15).

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Text description for Figure 3: Location of major ex situ conservation facilities for the Australian flora

A map of Australia split by state showing the locations of the major ex situ conservation facilities for the Australian flora:

  • In the Northern Territory:
    • George Brown Darwin Botanic Gardens conservation seed bank which has seedbanking and nursery production facilities
    • Alice Springs Desert Park which has seedbanking and nursery production facilities
  • In Western Australia:
    • Western Australian Seed Centre, Department of Biodiversity, Conservation and Attractions, Kensington, and Kings Park and Botanic Garden which has seedbanking, cryostorage, nursery production and micropropagation facilities
    • Forest Products Commission Seed Centre which has seedbanking and nursery production facilities
  • In South Australia:
    • The Australian Pastures Genebank, South Australian Research and Development Institute, which has seedbanking and nursery production facilities
    • the South Australian Seed Conservation Centre, Botanic Gardens and State Herbarium of South Australia (BGSH), which has seed banking and nursery production facilities
  • In Victoria:
    • The Australian Grains Genebank, Agriculture Victoria, which has seedbanking and nursery production facilities
    • The Victorian Conservation Seedbank, Royal Botanic Gardens Victoria, which has seedbanking, nursery production and micropropagation facilities
  • In Tasmania:
    • The Tasmanian Seed Centre, Sustainable Timber Tasmania, which has seedbanking and nursery production facilities
    • The Tasmanian Seed Conservation Centre, Royal Tasmanian Botanical Gardens, which has seedbanking, nursery production and micropropagation facilities
  • In New South Wales:
    • The National Seed Bank, Australian National Botanic Gardens, which has seedbanking, nursery production and micropropagation facilities
    • The Australian Tree Seed Centre, CSIRO, which has seedbanking and nursery production facilities
    • The Australian PlantBank, Australian Institute of Botanical Science, Royal Botanic Gardens and Domain Trust, which has seedbanking, cryopreservation, nursery production and micropropagation facilities
  • In Queensland:
    • The Brisbane Botanic Gardens Conservation Seed Bank, Brisbane Botanic Gardens, Mt Coot-tha, which has seedbanking and nursery production facilities

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Five plant pots each with a different species of Brassica being grown showing a variety of leaf morphologies
Brassica crop wild relatives incubated at 20°C to obtain leaf material for DNA extraction. (Photo: Pablo Gómez).

Wild edible plants (WEP) are, for the most part, uncultivated and have one or more edible parts. These plants can be well-known in the area where they are found and used, but often, the knowledge of how to use them has been lost.

Despite the large number of WEP reported (> 7,000 species, Diazgranados et al., 2020), only 30 are currently part of our diet, with more than 50% of our calorie intake based on wheat, rice, maize and sugar consumption (FAO, 2020). These few dominant crops have been bred over millennia, drastically narrowing their genetic diversity in comparison to wild species. Crop wild relatives (CWRs) are species closely related to crops, that are used as a source of genetic diversity to improve conventional crops. According to Castañda-Álvarez et al., (2016), there are more than one thousand CWRs registered, of which only 150 have their conservation status assessed (IUCN). In many cases, CWRs exhibit tolerance and resistance to biotic and abiotic stresses that can be transferred to crops via breeding techniques.

Many CWRs are threatened in the wild and need to be prioritised for ex situ collection to conserve their diversity. This is most pressing for endemic species, which may have unique genetic and phenotypic traits that could be lost if their habitat is vulnerable (e.g., to climate or land use change). Storing CWRs in seed or gene banks not only ensures their conservation but also makes material accessible for research into the traits that they possess.

Several collaborative projects between the Millennium Seed Bank (MSB) and its partners have undertaken seed collection of WEP, gathering more than 3,000 WEP (>15,000 accessions, Fig. 1). Furthermore, the MSB Partnership (MSBP) has collected and conserved CWRs from at least 25 crops (mainly pulses, cereals and forages listed in Annex I of the International Treaty on Plant Genetic Resources for Food and Agriculture) to investigate their characteristics and select key accessions for crop improvement purposes. Nearly 300 CWRs species (3767 accessions) were collected during 2010-2020 from 90 countries (Fig. 2).

Gradient heat map of the world to show areas where high and low numbers of populations of WEP accessions are conserved across the MSB
Figure 1: Distribution of wild edible plant accessions conserved ex situ at the MSB. The key shows the number of populations registered on a logarithmic scale. A text description for the map is available. (Image: RBG Kew).
Gradient heat map of the world to show areas where high and low numbers of populations of CWR accessions are conserved across the MSB
Figure 2: Distribution of CWR accessions conserved ex situ across the MSB, the magnified sections display UK and Israel as the top two countries with the most species conserved. The key shows the number of populations registered on a logarithmic scale. A text description for the map is available. (Image: RBG Kew.)

Despite the extraordinary work and extensive efforts of the institutions involved, WEP and CWRs continue to need urgent collection and conservation (Vincent et al., 2019, Maxted and Vincent, 2021). More than 50% of WEP and CWRs are still under-represented in genebanks and little knowledge is available for most of them. In response, a gap analysis was performed to identify key species that need to be prioritised for conservation, as well as regions or hotspots with a high number of WEP and CWRs not yet conserved across the MSBP. The objective is to ensure their long-term conservation, to increase the diversity and accessibility of collections.

Southeast Asia and Malesia, together with West and West-Central Tropical Africa were identified as the regions with the greatest number of WEP not yet conserved (Fig. 3). On the other hand, Southeast Asia and Latin America, more specifically Brazil, were identified as hotspots for CWRs that are not stored ex situ (Fig. 4). We recommend that the top five genera with missing collections for WEP (Elaeocarpus, Pinanga, Pandanus, Agave and Garcinia) and CWR (Artocarpus, Arachis, Solanum, Brassica and Prunus), together with other endemic taxa, should be targeted from the aforementioned regions, establishing new partnerships if needed, for conservation efforts. This will help secure the genetic diversity held in these species for current and future generations.

Gradient heat map of the world showing areas with high numbers of WEP populations not conserved. Described within article.
Figure 3: Distribution of wild edible plant occurances with no record of being conserved ex situ. The key shows the number of populations documented. The image is described in the text. (Image: RBG Kew).
Gradient heat map of the world showing areas with high numbers of CWR populations not conserved. Described within article.
Figure 4: Distribution of CWRs registered with no record of being conserved ex situ. The key shows the number of populations documented. The image is described in the text. (Image: RBG Kew).
References
  • Castañeda-Álvarez, N.P., Khoury, C.K., Achicanoy, H.A., Bernau, V., Dempewolf, H., Eastwood, R.J., Guarino, L., Harker, R.H., Jarvis, A., Maxted, N., Müller, J.V., Ramirez-Villegas, J., Sosa, C.C., Struik, P.C., Vincent, H. & Toll, J. (2016) Global conservation priorities for crop wild relatives. Nature plants. 2(4): 1-6. DOI: https://doi.org/10.1038/nplants.2016.22
  • Diazgranados, M., Allkin, B., Black, N., Cámara-Leret, R., Canteiro, C., Carretero, J., Eastwood, R., Hargreaves, S., Hudson, A., Milliken, W., Nesbit, M., Ondo, I., Patmore, K., Pironon, S., Turner, R. & Ulian, T. (2020) World checklist of useful plant species. RBG Kew. DOI: https://knb.ecoinformatics.org/view/doi:10.5063/F1CV4G34
  • Food and Agriculture Organization of the United Nations (2020) FAOSTAT Statistical Database. Rome: FAO. (accessed May 2022)
  • Genesys (2022). Data accessed through Genesys Global Portal on Plant Genetic Resources, www.genesys-pgr.org, (accessed Jan 2022).
  • Maxted, N., & Vincent, H. (2021) Review of congruence between global crop wild relative hotspots and centres of crop origin/diversity. Genetic Resources and Crop Evolution. 68(4): 1283-1297. DOI: https://doi.org/10.1007/s10722-021-01114-7
  • Seed Bank Database, SBD, (2021)(accessed Jan 2022)
  • POWO (2022) Plants of the World Online. Available at: https://powo.science.kew.org/ (accessed September 2021)
  • Vincent, H., Amri, A., Castañeda-Álvarez, N. P., Dempewolf, H., Dulloo, E., Guarino, L., Hole, D., Mba, C., Toledo, A. & Maxted, N. (2019). Modeling of crop wild relative species identifies areas globally for in situ conservation. Communications Biology. 2(1): 1-8. DOI: https://doi.org/10.1038/s42003-019-0372-z

Text description for Figure 1: Distribution of wild edible plant accessions conserved ex situ at the MSB.

A world map highlighting high numbers of wild edible plant population accessions held at the MSB from the United States, Mexico, Mali, Burkina Faso, Sudan, Tanzania, Botswana, South Africa, Madagascar, Italy, UK, China and Australia.

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Text description for Figure 2: Distribution of CWR accessions conserved ex situ across the MSB.

A world map highlighting high numbers of crop wild relative plant population accessions held at the MSB from the United States, Mexico, Ecuador, Chile, Ghana, Nigeria, Sudan, Uganda, Kenya, UK, Spain, Italy, Israel, Syria, Jordan, Turkey, Pakistan and Vietnam.

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Daucosma laciniata Engelm. & A.Gray, also known as Meadow Parasol or Meadow Daucosma, is a herb native to Texas and New Mexico in the United States (Lady Bird Johnson Wildflower Center, 2022; USDA, NRCS, 2022). It was first described to science in the Boston Journal of Natural History (volume 6, pg. 211) by Asa Gray in the article “An account of a collection of plants made by F. Lindheimer in the western part of Texas, in the years 1845-6, and 1847-8, with Critical Remarks, Descriptions of New Species” (IPNI, 2022). Digitised versions of the herbarium sheets from this collection are available through the Harvard University herbaria and RBG Kew herbaria.

Daucosma is a genus within the Apiaceae (Umbelliferae) family, with Daucosma laciniata being the only species of the genus (WCVP, 2022), making it taxonomically distinct. The species has had its genome sequenced as part of the Plant and Fungal Tree of Life (PAFTOL) project (available at: https://treeoflife.kew.org/specimen-viewer/10171).

Close up of Daucosma laciniata in flower, with the white flowers within the umbel and the divided bracts.
Daucosma laciniata. (Photo: Bill Carr, Lady Bird Johnson Wildflower Center).
Daucosma laciniata growing infront of a rock. The plant is in flower, showing the white flowers of the umbel.
Daucosma laciniata. (Photo: Bill Carr, Lady Bird Johnson Wildflower Center).

The species is an annual, flowering between June and July with white umbellifer flowers (Lady Bird Johnson Wildflower Center, 2022). Its current conservation status is G4 - Apparently Secure (NatureServe, 2022a), which means the species is at fairly low risk of extinction (NatureServe, 2022b).

One collection of the species is held at the Millennium Seed Bank (MSB), donated by the Lady Bird Johnson Wildflower Center in 2005. Since 2006 various germination tests have been performed on the collection without achieving significant germination. However, cut-tests at the end of these experiments revealed that the seeds were still alive, indicating that the perfect dormancy breaking conditions for the species remained elusive, rather than the collection being inviable . In 2019, Kirstine (Germination Assistant at the MSB) set up various tests based on a combination of the best conditions from previous germination test results and WORLDCLIM average monthly temperatures for the area the collection was made. Fast forward 504 days (72 weeks!) and one of the tests reached 92% germination (100% viability after accounting for empty and viable ungerminated seeds). The germination conditions consisted of 4 steps:

  • Firstly, 8 weeks cycling at 20°C for 12 hours, then 10°C for 12 hours with 12 hours of light each day.
  • Secondly, 16 weeks at 5°C with 12 hours of light a day.
  • Thirdly, 14 weeks cycling again at 20°C for 12 hours and then 10°C for 12 hours with 12 hours light a day.
  • And finally, 34 weeks cycling at 35°C for 12 hours and 20°C for 12 hours with 12 hours light a day.

This germination protocol still needs to be refined, although this will probably take another year and a half! However, it does show that the seeds are viable, and we have been able to break the dormancy of this particular collection which will allow for the propagation of the plants from seed in the future.

References

This month we spoke with Naomi Carvey, Millennium Seed Bank Partnership (MSBP) Data Warehouse Project Officer at the Millennium Seed Bank (MSB). Naomi has one of the most wide-reaching roles within the Seed Conservation team at the MSB and will have been in contact with many of you. So we thought we’d take the opportunity to introduce the person behind the email address!

Naomi out collecting as part of the UK National Tree Seed Project. (Photo: Naomi Carvey).
Naomi stood smiling holding a pole pruner and a bucket, wearing a hard hat on a misty hill top.
Naomi kitted out for seed collecting as part of the UK National Tree Seed Project. (Photo: RBG Kew).

What did you do before joining the MSB?

I’ve always had data and systems type roles in my career prior to joining Kew, but in very different industries. My first full-time job after leaving University was pricing flights and holidays for a big travel company, after that I started working in banking in the City of London, including working for several years at the Bank of England – the UK central bank. Before that, whilst I was still in full-time education I spent several years milking cows and helping with wedding catering during weekends and holidays – something quite different!

Why did you join the MSB and what interested you about the role?

While still working in banking and studying GIS I became interested in environmental datasets and applications. I heard about the new MSBP Data Warehouse project first, and through that rather stumbled across the wider work of the MSBP. I thought it sounded like a really interesting and dynamic place to work, and the role seemed to suit my experience and interests well.

What is your current role and what does a typical day look like for you?

I’ve been the MSBP Data Warehouse Project Officer for several years now, helping to gather, improve and secure data from seed collections across the MSB partnership, and to get the data back into the MSBP community. A typical day might involve checking and feeding back on a dataset from an MSB partner, and then uploading those data to the MSBP Data Warehouse online. I might also be updating IUCN red list data or species names, helping colleagues with data analysis or delivering data training.

Can you tell us about something you are proud to have achieved in your role?

Recently I have worked on the design and implementation of processes that allow us to receive herbarium vouchers from MSB partners who do not duplicate their seed collections at the MSB, but share full collection and curation data instead. I have found this work to be very enjoyable and rewarding. Duplicating herbarium vouchers at RBG Kew for seed collections stored elsewhere throughout the partnership really opens up fantastic possibilities for research and collaboration, as well as elevating seed collections to the highest quality standards, maximising collection utility and longevity.

What are some of the challenges you have faced, or think you might face in the future?

Increasingly I feel there is a gap between the fantastic wealth of seed collections, their associated data and new research done around the world, and getting those seeds and their associated knowledge back into living ecosystems. There is some amazing work being done across the MSBP in restoration and on new techniques in habitat management but currently issues around seed supply, a lack of germination / cultivation protocols and the difficulties transporting germplasm across borders are holding this work back. I think plant conservation should be a cycle of gathering collections and knowledge and putting them to practical use, but there are some bottlenecks at the moment. I’d love to get more involved in work to help tackle some of these issues.

Can you tell us about a favourite moment from your work for the MSBP?

Meeting colleagues from other seedbanks across the MSBP is always a highlight – we work with such a fantastically diverse group of people, and it is always great to learn about associated projects, not to mention discovering more about plants! The annual Seed Conservation Techniques (SCT) course is always special and I’ve also been lucky enough to visit some MSB partners in-country. Each visit has been really beneficial, and I have life-long memories of them all. To highlight one, the SCT course held at Cibodas Botanic Gardens in Indonesia in 2017 – the course had a significant impact on the future direction of the collaboration and a large number of people were able to join the training, which was very well organised and held in such beautiful surroundings. Through Arcadia and the Global Tree Seed Bank programme, the National Research and Innovation Agency (BRIN) - our MSB partner in Indonesia – is now working on collecting over 1000 species.

What advice would you give somebody regarding their data?

Concentrate on completing any mandatory fields first, and be prepared to invest a significant amount of time on your data at the beginning and during data collection – if you do a thorough job at the start you will save a lot of time and many headaches later on! You will also be able to maximise the longevity and potential future use of your collections. Don’t be afraid to ask for help from your colleagues at the MSB - we have a lot of varied experience and resources and we’re here to help.

If you could be one plant, what would you be and why?

I’d be a beautiful, craggy old Yew tree, Taxus baccata, a tree known for its cancer-fighting chemicals, toxicity and folkloric associations with death and resurrection which are sometimes thought of as portals to the underworld. In the UK ancient Yew trees are often found in churchyards and some pre-date the arrival of Christianity. I have fond memories of collecting T. baccata seeds for the UK National Tree Seed project - my first ever seed collecting trip for the MSB which was highly influential for me.

An old Yew tree growing on the top of a large rock, the roots of the tree are growing like a network over the rock
A Taxus baccata tree along the Rock Walk at Wakehurst. (Image: Jim Holden, RBG Kew).

How many seeds should I collect?

How many seeds to collect is a question of balance between making sure you can gather enough seeds for your intended use in the time you have available, while ensuring you do not harm the population from which you are sampling. In general, for a long-term conservation collection, you should aim to collect at least 10,000 potentially viable seeds, but not taking more than 20% of the available mature seed. Population data should be used to guide when more cautious collecting might be necessary, e.g., when making a collection from annual species that produce few seeds. The recommendation to collect 10,000 potentially viable seeds is based on the seed use figures in Table 1, which were calculated from collections of abundant species held at Kew, and is a starting point that can be adjusted for specific projects or collection events. You should also keep in mind the importance of maximising the genetic diversity captured, by collecting randomly from as many individuals (maternal lines) as possible.

Table 1: Indicative breakdown of seed numbers for different uses from a single collection (adapted from: MSBP Technical Information Sheet 02, RBG Kew).
Activity Indicative Seeds Required
Base collection kept in case of loss of wild population Ideally 500 seeds
Developing an effective germination protocol 100 seeds
Viability monitoring over the anticipated 200-year life span of the collection 650 seeds
Duplication at another bank for safety At least 1,150 seeds
Distribution to users 5,000 seeds (sufficient for 100 researchers to receive a 50 seed sample)
Future propagation and restoration projects At least 2,500 seeds for trialling
Total 9,900 seeds

The recommendation to collect 10,000 seeds might need to be adapted depending on the quality of the seed you are collecting. You will need to undertake a cut-test to verify the quality of seed from your sample population. Usually you need to cut-test ten seeds. If two of those ten are empty (20%), you can assume that 20% of the seeds you collect will be empty, therefore to collect 10,000 potentially viable seeds you would need to collect 12,000 seeds (10,000 x 1.2). You will then need to calculate again whether it is possible to collect this quantity while still ensuring you are taking no more than 20% of the available mature seed.

A fruit cut in half lying on the forest floor. The two of the seeds look healthy, the other two look like they are potentially inviable.
Cut-testing fruit in the field. (Photo: W. Stuppy).

In the case of rare and threatened species, what is the minimum sized collection that is useful for a seed bank? There is no absolute threshold but ideally you should aim for a collection of 500 seeds. However, this will not always be possible and it is important to consider other factors, for example:

  • does the safe-collecting limit need to be lower?
  • how many seeds are available?
  • what are the intended end uses of the seed?

In cases where it is not possible to collect sufficient seed in a single visit, repeating collections over multiple years may be an alternative option.

This text was adapted from Technical Information Sheet 02 - Assessing populations, (RBG Kew, 2022).

Three people, each holding a paper bag, collecting dandelion heads in seeds.
Seed collecting in the field. (Photo: Hanna Oldfield).

Recent publications from across the MSBP:

  • Carta, A., Fernández-Pascual, E., Gioria, M., Müller, J.V., Rivière, S., Rosbakh, S., Saatkamp A., Vandelook F. & Mattana, E. (2022) Climate shapes the seed germination niche of temperate flowering plants: a meta-analysis of European seed conservation data. Annals of Botany. mcac037. DOI: https://doi.org/10.1093/aob/mcac037
  • Carta, A., Mattana, E., Dickie, J. & Vandelook, F. (2022) Correlated evolution of seed mass and genome size varies among life forms in flower plants. Seed Science Research. 32: 46-52. DOI: https://doi.org/10.1017/s0960258522000071
  • Hay, F.R., Davies, R.M., Dickie, J.B., Merritt, D.J. & Wolkis, D.M. (2022) More on seed longevity phenotyping. Seed Science Research. 1-6. DOI: https://doi.org/10.1017/S0960258522000034
  • Hardstaff, L.K., Sommerville, K.D., Funnekotter, B., Bunn, E., Offord, C.A. & Mancera, R.L. (2022) Myrtaceae in Australia: Use of Cryobiotechnologies for the Conservation of a Significant Plant Family under Threat. Plants. 11(8): 1017. DOI: https://doi.org/10.3390/plants11081017
  • Pirie, M.D., Blackhall-Miles, R., Bourke, G., Crowley, D., Ebrahim, I., Forest, F., Knaack, M., Koopman, R., Lansdowne, A., Nürk, N., Osborne, J., Pearce, T.R., Rohrauer, D., Smit, M. & Wilman, V. (2022) Preventing species extinctions: A global conservation consortium for Erica. Plants, People, Planet. First view. https://doi.org/10.1002/ppp3.10266
  • Rodríguez-Zúñiga, J., Flores-Ortiz, C.M., González-Guillén, M.d.J., Lira-Saade, R., Rodríguez-Arévalo, N.I., Dávila-Aranda, P.D. & Ulian, T. (2022) Cost analysis of seed conservation of commercial pine species vulnerable to climate change in Mexico. Forests. 13: 539. DOI: https://doi.org/10.3390/f13040539
  • Tiloca, G., Brundu, G. & Ballesteros, D. (2022) Bryophyte spores tolerate high desiccation levels and exposure to cryogenic temperatures but contain storage lipids and chlorophyll: understanding the essential traits needed for the creation of Bryophyte spore banks. Plants. 11:1262. DOI: https://doi.org/10.3390/plants11091262

MSBP Technical Information Sheet Update

The Technical Information Sheets available on the Resources page of the MSBP website have recently been updated and republished. The Technical Information Sheets cover various aspects of seed conservation practices and facilities for example:

As part of these updates all the pdf's have been made web accessible. If you have any issues using the Technical Information Sheets, please contact samara@kew.org .




Issue 2: March 2022

This issue we hear about field, lab and training activities from Mozambique, Australia and Indonesia. We interview Victoria Wilman, Seed Conservation Programme Manager at SANBI (South African National Biodiversity Institute) and this month's species profile comes from Indonesia - the Begonias native to the Mount Slamet region. We also check back in with the Seeds of Hope featured in the news section of the December issue of Samara.

A germinated seedling of I. dunensis, the seed standing above the sand on a cream hypocotyl and stem and cotyledons emerging from the top of the seed
Icuria dunensis seedling, Mozambique. Photo: Cacilda João Chirinzane Manhiça.
Flowering spike of Calochilus paludosus with two buds and one green and red flower showing the characteristic red hairs
Calochilus paludosus, Australia. Photo: Dan Duval.
Two flowers each with six white petals with a purple streak at the centre of the flower
Aristea spiralis, South Africa. Photo: Victoria Wilman.
Two female flowers each with three white tepals, lobed yellow stigmas and a light pink, 3-winged ovary
Begonia muricata, Indonesia. Photo: M Efendi.

Nationally endemic plants are very important in conservation planning as their survival depends on only one country. Icuria dunensis (Fabaceae) is not only an endemic species to Mozambique, but also one of Mozambique’s five endemic genera (Darbyshire et. al., 2019a). It is listed as Endangered on the IUCN Red List of threatened species (Darbyshire et. al., 2019b) and its distribution is limited to a stretch of about 200 km along the Mozambican coast between Nacala (Nampula province) and Moebase (Zambezia province).

Icuria trees can grow to 40 m tall. They are dominant in patches of dry coastal forest, sometimes forming pure stands (Burrows et. al., 2018). The fruits are velvety woody pods about 10 cm long, with only 1-3 large seeds inside.

Importance of the tree for communities

Icuria trees are valued by local communities. For example, in the Mulimone Icuria forest in Nampula province, trees are debarked for canoe making (Fig. 1) and the wood is used for construction and firewood. However, trees are also killed by ring-barking a section of the trunk to clear the land for agriculture (Fig. 1). In addition, clearance of vegetation during the mineral sand mining occurring in this region, threatens the remaining patches of these Icuria forests.

A branched tree trunk with a long section lighter in colour where the bark has been removed The base of a tree trunk, red in colour with streaks of red sap running down it. There is a wide indentation where a strip of bark has been removed around the trunk
Figure 1: Local communities debark long sections of Icuria trunk to make canoes, killing large branches (left) and ring-bark a narrow strip of the trunk to kill the tree during land clearance for agriculture. The Icuria tree releases a red sap (right). Photos from Mulimone Icuria forest in Nampula Province. (Photos: Cacilda João Chirinzane Manhiça).
Orthodox or Recalcitrant Icuria dunensis seeds?

In its habitat we noticed many Icuria trees completely without, or with very low numbers of, mature fruits. In order to guarantee enough quality seeds of Icuria for restoration purposes there is an urgent need for ex situ conservation by seed banking. Very little is known about the seeds of this species. They are large and fleshy with a thin seed coat, which suggests they may be recalcitrant.

Preliminary seed tests were conducted using seeds collected by Kenmare in Nampula Province, Larde district, in January 2019. The tests were carried out at the IIAM-CIF laboratory (Fig. 2). To determine the seed moisture content, fresh seeds were cut into small pieces using secateurs and divided into two samples of 10 grams. The samples were dried in the oven at 130°C for an hour. The fresh seed weight was about 87 seeds/kg, with a moisture content of 34.5% (IIAM-CIF, 2020).

A pile of round disc like seeds on a laboratory table, Zélia, Milton and Horácia are stood around the table holding secateurs
Figure 2: Preparing I. dunensis seeds for laboratory tests: IIAM staff from left to right: Zélia Malate, Milton Zavale and Horácia Boene. (Photo: Cacilda João Chirinzane Manhiça).
Germination room

124 seeds of I. dunensis were sown in sterilised sand in the germination room, with a temperature between 25-30°C, 24 hours photoperiod and air relative humidity between 60-80%. The seeds started germinating between day 3 and day 21. The germination was epigeal, with the cotyledons forcing above the surface of the germination medium as the seedlings grew (Fig. 3).

The 91% germination result was very encouraging and exceeded our expectations. However, this was just the start of our Icuria seed conservation research. Further tests are required to check if the seeds are indeed sensitive to desiccation, and to determine how germination responds to drying (Gold and Hay 2014).

Round seeds protruding a couple of centimeters above a tray of sand A germinated seedling of I. dunensis, the seed standing above the sand on a cream hypocotyl and stem and cotyledons emerging from the top of the seed
Figure 3: Icuria dunensis seeds germinating in sterilized sand, showing epigeal germination. (Photo: Cacilda João Chirinzane Manhiça).
Production of seedlings

The seedlings were pricked out, transplanted into polyethylene pots and placed in the nursery under shade. Five months later, the seedling survival rate was 100% and the seedlings reached 30-40 cm tall (Fig. 4). This is encouraging as it shows that if fresh seeds are available, it may be easy to produce seedlings for restoration and ex situ conservation.

6 rows of Icuria dunensis saplings, each sapling in an individual round planter
Figure 4: Icuria dunensis seedlings, 30-40cm tall in the IIAM-CIF nursery, July 2019. (Photo: Cacilda João Chirinzane Manhiça).
Next steps
  • Test if the seeds are desiccation-sensitive using the seed coat ratio model and seed storage behaviour protocols (Gold and Hay 2014);
  • Send duplicate seeds to the MSB for further seed testing including germination testing after banking;
  • Reinforce/strengthen the in situ conservation and restoration programme for Icuria in partnership with Kenmare and other potential partners.

This work was carried out under the MoU between IIAM-Kenmare to restore Icuria dunensis and implement in situ conservation initiatives. To be able to continue our Icuria seed research we appreciate the support of the Global Tree Seed Bank and Threatened Biodiversity Hotspots programmes that are funding Mozambique’s National Seed Conservation Programme at IIAM.

References
  • Burrows, J.E., Burrows, S.M., Lötter, M.C. & Schmidt, E. (2018) Trees and shrubs Mozambique. Publishing Print Matters (Pty) Ltd, Noordhoek, Cape Town.
  • Darbyshire, I., Timberlake, J., Osborne, J., Rokni, S.; Matimele, H., Langa, C., Datizua, C., de Sousa, C., Alves, T., Massingue, A., Hadj-Hammou, J., Dhanda, S.; Shah, T. & Wursten, B. (2019a) The endemic plants of Mozambique: diversity and conservation status. Phytokeys. 136: 45-96. DOI: https://doi.org/10.3897/phytokeys.136.39020
  • Darbyshire, I., Massingue, A.O., Osborne, J., De Sousa, C., Matimele, H.A., Alves, M.T., Burrows, J.E., Chelene, I., Datizua, C., Fijamo, V., Langa, C., Massunde, J., Mucaleque, P.A., Rokni, S. & Sitoe, P. (2019b) Icuria dunensis. The IUCN Red List of Threatened Species 2019: e.T136532836A136538183. DOI: https://dx.doi.org/10.2305/IUCN.UK.2019-2.RLTS.T136532836A136538183.en (Accessed on 02 February 2022)
  • Gold, K. & Hay, F. (2014) Identifying desiccation-sensitive seeds. Technical Information Sheet 10. Millennium Seed Bank Partnership. Royal Botanic Gardens Kew. PDF: https://brahmsonline.kew.org/Content/Projects/msbp/resources/Training/10-Desiccation-tolerance.pdf
  • IIAM-CIF (2020, Report). Relatório sobre testes de qualidade de sementes de Icuria dunensis fornecida pela Kenmare-REHAB.

The work of the Australian Seed Bank Partnership (ASBP) is built on collaboration. Our relationship with Kew’s Millennium Seed Bank Partnership (MSBP) has been instrumental to our success in building capacity to deliver seed banking projects over the past 20 years. Following the devastating 2019–20 Australian bushfires, the MSBP reached out once more to help us secure our threatened flora. Thanks to a very generous offer of support from the UK Foreign, Commonwealth and Development Office (FCDO) in February 2020, we have been able to undertake an emergency collecting programme in impacted areas during the first season post-fire.

Target species for this programme were selected based on the severity of bushfire impact, their threatened status, and their potential to provide critical ecosystem services that support Australia’s native wildlife. Our partners made 47 collections of 42 taxa and delivered 164 germination trials of 93 taxa, representing flora from every Australian state and territory. On Kangaroo Island, two rare orchids (pictured) were located and banked by the South Australian Seed Conservation Centre. The funding provided for this programme has also enabled us to propagate 10 priority plants in Western Australia, Victoria and South Australia for planting in seed production areas and botanic garden displays.

Yellow orchid flower of Caladenia transitoria subspecies isolata
The shy gremlin orchid (Caladenia transitoria subsp. isolata) was previously only known from a single historic record, but in spring 2020 two small populations were discovered after fire. (Photo: Dan Duval).
Flowering spike of Calochilus paludosus with two buds and one green and red flower showing the characteristic red hairs
A small population of the endangered swamp bearded orchid (Calochilus paludosus) was also located on Kangaroo Island in spring 2020. (Photo: Dan Duval).

To date, ASBP partners have also undertaken rapid flora assessments for 11 bushfire affected flora to ascertain habitat condition, initial fire response, species abundance, pests and disease presence and overall fire impact. To ensure standardised data collection across multiple states, our partners developed a methodology for species monitoring. This programme funded the printing of dedicated field books to capture this information. The assessments have provided seed banks with crucial information about species recovery post-fire. The data will be uploaded to botanic gardens databases and made available to those involved in research, conservation, and bushfire recovery actions to monitor recovery over time and inform future seed collection priorities.

Despite unforeseen project delays due to COVID restrictions, flooding, and impacts of the current La Niña event, the Partnership has adapted to ensure an ongoing commitment to ex situ conservation of Australian flora. This includes adjusting our species target lists to collect other accessible priority species, postponing our activities to the 2021–22 summer season, and establishing seed orchards from previous collections where species cannot be feasibly located.

The work our partners have delivered under this project, and many like it, mean thousands of native species are now secured in seed banks throughout Australia. We hope our efforts and the ongoing interest from beyond our shores will inspire people to learn more about protecting our native plants in our vast and varied continent.

The Millennium Seed Bank Partnership (MSBP) has received the World Wide Biodiversity Conservation Award at the 16th BBVA Foundation Awards ceremony in Madrid, 30th November 2021. Each year the BBVA Foundation, the corporate social responsibility arm of the BBVA financial services group, hosts an award ceremony recognising those working for Biodiversity Conservation. Three awards are given, one for biodiversity conservation in Spain, one for knowledge dissemination and communication in biodiversity conservation in Spain, and one for worldwide biodiversity conservation.

It is this global category that the Millennium Seed Bank Partnership (MSBP) has won “for its extraordinary contribution to the preservation of the world’s plant biodiversity, through the creation of a seed bank holding 2.5 billion samples of plants from 190 countries”. The judges considered the MSBP to be an exemplary initiative which reflects how cooperation without borders can advance nature conservation worldwide and successfully address the central challenge of preserving biodiversity. They were particularly impressed by the MSBP's training capacity, a key element of the partnership which allows best practice, technology and ideas to be shared across the network.

Dr Elinor Breman, lead of the MSBP Initiative at Kew, travelled to Madrid to accept the award, with colleagues Dr Aisyah Faruk who coordinates our work across Europe, and Dr Tiziana Ulian, who works on diversity and livelihoods.

A person standing on a stage making a presentation to an audience
Dr Elinor Breman, Senior Research Leader at the Millennium Seed Bank, addresses the audience at the 16th BBVA Foundation Awards ceremony in Madrid on the 30th November 2021. (Photo: RBG Kew).

The award also provided 250,000 Euros to further the work of the MSBP. In the spirit in which the award was given, this funding is being used to continue our training programme and conserve critically endangered plants, bringing them back from the brink of extinction.

The funding will enable 10 people to undertake Technical Attachments at the Millennium Seed Bank and ~12 people to attend our flagship Seed Conservation Techniques course in 2022. The remaining funds will support a new four-year project to conserve 50 plant species designated as critically endangered (CR) by the global IUCN Red List.

Mind the Gap - What CR species are conserved?

The species to be conserved have been identified through an internal review of MSBP holdings of CR species, which identified CR species missing from collections as well as those whose collections could be improved (increase in the number of seeds from a population or the number of populations conserved). Ten countries will be involved in not only collecting these CR species, but drawing together the known information about them, developing propagation protocols and then feeding information and plants back into in situ conservation efforts.

The award engraved with the text Biodiversity Conservation Awards, BBVA Foundation Worldwide Award for Biodiversity Conservation, Royal Botanic Gardens, Kew, Madrid, November 30 2021
BBVA Foundation Worldwide Award for Biodiversity Conservation. (Photo: RBG Kew).

The transfer of seed conservation knowledge through the Millennium Seed Bank Partnership (MSBP) over the last two decades has been reviewed by Way & Breman (2021). They show how the MSBP’s international training programme has successfully translated research knowledge into practical standards and guidelines. They also illustrate how the MSBP continues to develop decision-support tools such as the Seed Recalcitrance Predictor and the MSBP Data Warehouse and shares best practice in seed conservation through technical publications and partnership newsletters.

For 2022 we are hoping to run our Seed Conservation Techniques training course onsite at the Millennium Seed Bank (MSB) for the first time since 2018. The proposed dates include a week of online theory from September 26th- 30th, followed by practical hands-on sessions from October 9th - 22nd. Demand is likely to be high, so if you would like further information please contact MSBTraining@kew.org as soon as possible. The deadline for expressions of interest is April 30th.

A group of people wearing lab coats and gloves stood around a sink washing fruits in a sieve
Participants of the Seed Conservation Techniques Course, 2018, cleaning wet fruit in the MSB cleaning lab. (Photo: RBG Kew).
References

In the December edition of Samara, we featured a news story on receiving seeds from the hibaku jumoku (survivor trees) from the Green Legacy Hiroshima Initiative. The trees survived the atomic bomb dropped on the city of Hiroshima, Japan, on August 6, 1945, at the end of the Second World War and are now grown around the world as a symbol of peace and hope.

Many of the species we received from the Hiroshima survivor trees need cold stratification treatment and are still in the fridge. Some however, needed no pre-treatments. They were sown straight away and put into our glasshouse at a cosy 22°C.

The Cinnamomum camphora (Camphor Tree) seeds were first to the mark, with all three germinating (the first germinating after 4 weeks). They will soon be potted into airpots and as they grow (and toughen up), acclimatised to cooler conditions. I look forward to the day when these survivors will be planted out in the gardens at Wakehurst!

In the foreground three seedlings growing in a container filled with soil, with other plants in pots in the background
Three seedlings of Cinnamomum camphora from the Green Legacy Hiroshima Initiative, successfully germinated in the nursery at Wakehurst. (Photo: Eliana Van Der Schraft).

Notes on the Begonias of Mt. Slamet, Central Java, Indonesia

Muhammad Efendi, Abidin Ibrahim, Dadang Sunandar, Intani Quarta Lailaty & Vandra Kurniawan (Cibodas Botanic Gardens, Research Center for Plant Conservation and Botanic Gardens, National Research and Innovation Agency)

Begonia is known as an ornamental plant by many plant enthusiasts. It has an interesting shape and colour to the leaves, and also has many varied colours of flowers (Tebitt, 2005). Several species of Begonia have other uses, for example B. comestibilis is used for food (Thomas et al., 2011), B. multangula as a spice, B. muricata (Hartutiningsih et al., 2009; Putri et al., 2019), B. medicinalis (Ardi et al., 2019) and B. baliensis (Hartutiningsih et al., 2018) were reported as potential medicines. These species contain antimicrobial compounds with the potential to be developed as medicinal raw materials.

Mount Slamet is known as habitat of native Begonia’s in Java. A total of five native Begonia have been reported in this area in a previous study (Efendi, 2019). A seed collecting expedition was conducted in the Mount Slamet forest area, Central Java, Indonesia, to increase the number of Begonia seed collections which are stored in the Cibodas Botanic Garden’s Seed Bank. A total of six species of native Begonia were found and collected within the South Slope area of Mount Slamet, namely B. multangula, B. longifolia, B. areolata, B. atricha, B. isoptera and B. muricata. A new location of Begonia isoptera was added to its range distribution from this expedition. Thus, as many as 40% of Java's native species of Begonias are now stored in Cibodas Botanic Garden’s Seed Bank and duplicated at the Seed Bank of the Center for Plant Conservation and Botanic Gardens. Unfortunately, there is a decreasing trend of the B. atricha population in Mt. Slamet caused by over exploitation for ornamental plants.

Two leaves with lobed margins and pointed lobe tips
Begonia multangula. (Photo: M. Efendi).
Underside of stem showing the alternate leaf arrangement and asymmetrical leaf base, white buds are hanging from the leaf axils
Begonia longifolia. (Photo: M. Efendi).
Two female flowers each with three white tepals, lobed yellow stigmas and a light pink, 3-winged ovary
Begonia muricata. (Photo: M. Efendi).

Orange-red, pubescent flower buds at the stem apex
Begonia areolata. (Photo: M. Efendi).
Two dark green asymmetrical leaves with serrate margins
Begonia atricha. (Photo: M. Efendi).
A stem with leaves in an alternate arrangement, a pale green developing fruit and inflorescences of pinky cream male flowers coming from the leaf axils
Begonia isoptera. (Photo: M. Efendi).
References
  • Ardi, W.H., Zubair, M.S., Ramadanil, P. & Thomas, D.C. (2019) Begonia medicinalis (Begoniaceae), a new species from Sulawesi Indonesia. Phytotaxa. 423(1): 41-45. DOI: https://doi.org/10.11646/phytotaxa.423.1.5
  • Efendi, M. (2019) Begonia alam Kebun Raya Baturaden. Prosiding Seminar Nasional Masyarakat Biodiversitas Indonesia. 5(1): 13-17. Available at: https://smujo.id/psnmbi/article/view/3216
  • Siregar, H-M., Purwantoro, R.S., Praptiwi, P. & Agusta, A. (2018) Antibacterial potency of simple fractions of ethyl acetate extract of Begonia baliensis. Nusantara Bioscience. 10: 159-163. Available at: https://smujo.id/nb/article/view/2726
  • Putri, N.H.S., Nurdiwiyati, D., Lestari, S., et al. (2019) Aktivitas antibakteri ekstrak tangkai dan daun Begonia multangula Blume. terhadap Porphyromonas gingivalis. Jurnal Biologi Universitas Andalas. 7(1): 51-58.
  • Thomas, D.C., Ardi, W.H. & Hughes, M. (2011) Nine new species of Begonia (Begoniaceae) from South and West Sulawesi, Indonesia. Edinburgh Journal of Botany. 68(2): 225–255. DOI: https://doi.org/10.1017/S0960428611000072

Our collaboration with SANBI (The South African National Biodiversity Institute) was one of the first under the global Millennium Seed Bank Partnership (MSBP). In over 20 years of working together the MSBP/SANBI team have made more than 6,500 collections, covering more than 4,350 species.

We spoke to Victoria Wilman (Seed Conservation Programme Manager) to learn more about her team's incredible contribution and their hopes and plans.

Photo montage of the SANBI MSBP team
The SANBI MSBP team, left to right: Victoria Wilman, Fergy Nkadimeng, Sibahle Gumede, Ntsakisi Masia, Naomi Mdayi, Thembeka Malwane and Jacqui James. (Photo: MSBP SANBI).

What influenced you to work in seed conservation?

I used to spend hours hiking in the mountains and visiting South Africa’s National Parks. I loved plants and collected seeds wherever I could. When my grandmother passed away, she left me a small legacy, just enough to kick start my first year of studies. I wanted to do nature conservation, but knowing about the many rare and endangered plants, I decided to study horticulture, to be able to grow these plants to restore them back into the wild. After working in the Eastern Cape and completing my honours degree in Botany a few years later, I applied for a seed collecting position in Cape Town and started at SANBI in 2012. I have loved every minute of it!

How has the global pandemic influenced your work and personal life?

The pandemic stopped all of our fieldwork, and our teams collecting seeds around the country were grounded although some colleagues who live in remote areas continued to collect around their home villages. I have an amazing team, and despite all the Covid challenges, they continued to work extremely hard and remained dedicated to the MSBP project, with the result that we are not far behind our collecting targets.

How do you think SANBI has developed its commitment to Seed Conservation over the past 20 years?

SANBI's commitment to seed conservation is very strong, the seed room at Kirstenbosch National Botanical Gardens (NBG) has changed from a commercial operation to a Seed Conservation Unit servicing the whole of SANBI.

A Seed Conservation Programme is being developed which includes infrastructure for South Africa’s first wild plant seed bank at Kirstenbosch NBG, including dry rooms, a -20°C; freezer, germination and seed processing labs, all due to be completed in early 2023. This exciting project brings together a lifetime’s ambition for me.

The seed conservation programme is well integrated into SANBI’s other directives and many of SANBI’s permanent staff have come through the MSBP programme. Our seed collectors are stationed at four of SANBI’s Botanical Gardens and work together with horticulturists and curators. We work closely with the Threatened Species Programme and Custodians of Rare and Endangered Wildflowers (CREW) who manage volunteers monitoring threatened species, and we also train them in seed collecting for the project.

A map of South Africa highlighting the collecting areas covered by each of the MSBP teams, and their respective locations
Map showing the collecting areas of the MSBP teams. Detailed alternative text for this map is available at the bottom of the article. (Photo: Victoria Wilman).

You have a strong Seed Conservation team at SANBI. Can you tell us a bit about the current team of researchers, their strengths and aspirations?

The team is comprised of myself, four seed collectors, one seed processing coordinator with support staff and an intern. We are finalizing two new projects at Kirstenbosch and Karoo Desert NBG which will add 3 more to the team.

Seed Collectors:

  • Naomi Mdayi – Kirstenbosch NBG (Western Cape)
  • Sibahle Gumede – Kwelera NBG (Eastern Cape)
  • Fergy Nkadimeng – Pretoria NBG (Gauteng, Freestate, North West)
  • Ntsakisi Masia – Thohoyandou NBG (Limpopo, Mpumulanga)
  • Jacqui James – Intern Kirstenbosch
  • New Project - Karoo Desert NBG (Northern Cape), succulent poaching
  • New Project – Kirstenbosch NBG, Restoration Unit, succulent poaching

Seed Processing at Kirstenbosch NBG:

  • Thembeka Malwane – seed processing coordinator
  • Georgina Wilkinson – seed cleaning and packaging shipments
  • Deon Smith – Administration (requests for material and other Gardens work)
  • Siyabonga Magladla – seed cleaning
  • Patrick Kettledas – seed cleaning and collecting

Due to the vast and varied terrain of South Africa the scope of our work is immense. The seed collectors (all female) spend extended periods away and are always accompanied in the field for their security. They are dedicated and knowledgeable. Amongst their many talents they also train students and volunteers, as well as coordinate the field trips.

Sibahle Gumede, from Kwelera NBG in the Eastern Cape shared her thoughts:

My aspirations are to see a South Africa that no longer has plants which are threatened, to have independent volunteers to carry out the MSBP vision, and universities, landowners and environmental stakeholders taking ownership for seed conservation. And lastly to see satellite seed banks for each province, to be able to curate their endemic seeds.

Your team engages with volunteers across South Africa, what are the key factors that make these partnerships work and are there any challenges?

Most of our stakeholders, partners, and volunteers understand and have experience of the effects of not preserving indigenous species. Our partnership with them is symbiotic and mutual and we are extremely grateful for all the help we receive from volunteers who help find target species and even collect seeds. We occasionally have difficulties getting hold of landowners for site access and they may prohibit us from collecting, and we can sometimes be viewed as a threat that exploits vegetation to give away seeds to another country, but we work hard at spreading the conservation message and more often than not receive cooperation and enthusiastic assistance.

You alone have been involved with 500 of the collections made by the MSBP/SANBI team. Do you have any collection highlights?

One of my favourite experiences is collecting from Silvermine Nature Reserve after a huge fire in 2015, which many viewed as devastating. We knew it was just the beginning of another cycle in nature and we discovered many species which were previously not present and some that come up en masse after a fire, like the beautiful Aristea spiralis, Disa racemosa and Erica amoena.

Two flowers each with six white petals with a purple streak at the centre of the flower
Aristea spiralis resprouting after the fire at Silvermine Nature Reserve. (Photo: Victoria Wilman).
Small woody shrub with whorls of green leaves and yellow daisy like flowers growing against a rocky background
Euryops indecorus. (Photo: Nick Helm).
Lots of green shoots and purple flowers emerging amongst burnt branches and bare ground
Life returning to the burned landscape at Silvermine Nature Reserve after the autumn rains. (Photo: Victoria Wilman).

Another great memory was collecting Euryops indecorus, a Critically Rare species known from only one site on cliff faces above Rooiels in the Western Cape. It took a whole day to get to and climb to the top of the mountain where this plant occurs and what a joyful and exciting feeling to find the species in flower and in seed, and to know that it is now secured in the seedbank.

We know that SANBI is pivotal in driving in situ species conservation and recovery, are there any key initiatives for which your seed collections will be used?

In partnership with MSBP, CREW, Kirstenbosch NBG and Stellenbosch Botanical Garden we are working on the Marasmodes undulata project. This species is classified as Critically Endangered and only occurs at a single locality. When last monitored, only three plants were present. The site has since burned, leaving no plants left in the wild. Seeds originally collected in 2005 were taken out of the seedbank and germinated at Kirstenbosch NBG and the Stellenbosch University BG. These plants are the basis for restoring this species, and these, and additional cuttings taken from them, will be planted into the wild this Autumn. It shows how important having ex situ collections can be.

How do you see the role SANBI plays in Southern Africa?

SANBI plays a critical role in the conservation arena, it leads and coordinates conservation research, and monitors and reports on the state of biodiversity in South Africa while also managing 10 Botanical Gardens that create awareness and showcase South Africa’s plants and provide education programs for learners. SANBI pilots wild plant seed conservation through our MSBP and seed conservation programme and highlights its importance through training programmes to many other organisations.

What do you see as the future challenges you are facing in your work?

Succulent poaching is a major problem, with plants exported for their ornamental value and the safety of my teams in the field is always a concern. Managing the new seed bank and developing a good seed science programme will be a new challenge, but a wonderful and exciting one.

You recently attended the remote Seed Conservation Techniques (SCT) course. How did you find it, and do you have any plans to further develop your current in-country training programme for staff and volunteers?

It was a great course; I really enjoyed it and learned so much. It was well organised with many resources. We are embarking on a staff training day every month, covering one aspect of the SCT training, and we will use the resources and videos from the course.


Detailed alternative text for map

A map of the provinces of South Africa showing the location of the different MSBP teams and the National Botanic Gardens (NBG) they’re based at, plus their respective collecting areas. The MSBP team based at Thohoyandou NBG in Limpopo cover collecting areas within the provinces of Limpopo and Mpumalanga. The MSBP team based at Pretoria NBG in Gauteng province cover collecting areas within the provinces of Gauteng, North West and Free State. The MSBP team based at Kwelera NBG cover collecting areas within the Eastern Cape and KwaZulu-Natal provinces. There are two MSBP teams within the Western Cape, one at the Karoo Desert NBG and one at Kirstenbosch NBG, these cover collecting areas within the Northern Cape and Western Cape provinces.

How do you perform a cut test on very small seeds, especially when in the field?

There are two elements to handling small seeds. One is to make sure they stay in place and don’t fly away, the other is to use the right tool for cutting.

We suggest the use of adhesive tape, by taking a strip of the tape and creating a loop (with the adhesive side facing outwards to fix the seeds), and then using small scissors to cut the seeds (either on the tape surface or by cutting through the tape with seed in place) (Fig. 1). Experienced collectors may also use their fingernail for cutting. It is always helpful to use a hand lens or microscope when cut testing very small seeds. Very tiny seeds, such as those of orchids, can’t be effectively cut tested.

A loop of adhesive tape over a finger, with several small seeds stuck to the tape. A small pair of scissors are being used to cut the small seeds
Figure 1: Small seeds stuck to the outside of a loop of adhesive tape, small scissors can then be used to cut the tape. (Photo: RBG Kew).

Recent publications from across the MSBP:

  • Diantina, S., McGill, C., Millner, J., Nadarajan, J., Pritchard, H.W., Colville, L. & Clavijo McCormick, A. (2022) Seed viability and fatty acid profiles of five orchid species before and after ageing. Plant Biology. 24: 168-175. DOI: https://doi.org/10.1111/plb.13345
  • Faruk, A., Papikyan, A. & Nersesyan, A. (2021) Exploring effective conservation of charismatic flora: orchids in Armenia as a case study. Diversity. 13(12): 624. DOI: https://doi.org/10.3390/d13120624
  • Liu, D., Cai, J., He, H., Yang, S., Chater, C.C.C. & Yu, F. (2022) Anemochore seeds harbour distinct fungal and bacterial abundance, composition, and functional profiles. Journal of Fungi. 8(1):89. DOI: https://doi.org/10.3390/jof8010089
  • Kallow, S., Garcia Zuluaga, M., Fanega Sleziak, N., Nugraha, B., Mertens, A., Janssens, S.B., Gueco, L., Valle-Descalsota, M.L., Dang Vu, T., Toan Vu, D., Thi Li, L., Vadeloo, F., Dickie, J.B., Verboven, P., Swennen, R. & Panis, B. (2022) Drying banana seeds for ex situ conservation. Conservation Physiology. 10: coab099. DOI: https://doi.org/10.1093/conphys/coab099
  • Pence, V.C., Meyer, A., Linsky, J., Gratzfeld, J., Pritchard, H.W., Westwood, M. & Bruns, E.B. (2022) Defining exceptional species - A conceptual framework to expand and advance ex situ conservation of plant diversity beyone conventional seed banking. Biological Conservation. 266: 109440. DOI:https://doi.org/10.1016/j.biocon.2021.109440
  • Pence, V.C., Bruns, E.B., Meyer, A., Pritchard, H.W., Westwood, M., Linsky, J., Gratzfeld, J., Helm-Wallace, S., Liu, U., Rivers, M. & Beech, E. (2022) Gap analysis of exceptional species - Using a global list of exceptional plants to expand strategic ex situ conservation action beyond conventional seed banking. Biological Conservation. 26: 109439. DOI: https://doi.org/10.1016/j.biocon.2021.109439
  • Visscher, A.M., Boatfield, M., Klak, C., Yeo, M., Pearce, T.R., Wilman, V., Mdayi, N., Gumede, S. & Pritchard, H.W. (2022) Physiological seed dormancy of Ruschia imbricata and Ruschia uitenhagensis (Aizoaceae) is broken by dry heat and unaffected by seasonality. South African Journal of Botany. 147: 457-466. DOI: https://doi.org/10.1016/j.sajb.2022.02.007

Important notice regarding the Seed Information Database

A recent assessment of the Seed Information Database (SID) for compliance with UK legislation on website accessibility (Public Sector Bodies [Websites and Mobile Applications] Accessibility Regulations 2018; themselves aligned with the Web Content Accessibility Guidelines (WCAG) 2.1 at the AA standard) found the web interface deficient in several respects. Regrettably, due to RBG Kew’s need to prioritise its limited resources, we will be unable to remedy the faults; and the SID web interface will be taken down at the end of this financial year (March 2022).

However, we will continue to update SID's data content internally; and this will continue to be available to external users soon as flat files from RBG Kew's FTP (file transfer portal) site.

Please also note that for many years we have shared much of SID's seed functional trait data via the TRY Plant Trait Database, which facilitates bulk data downloads; and we intend to continue periodic updates of that content.

In the future, as far as possible, we plan to deliver species' seed trait information via Kew's 'Plants of the World Online' portal, which should be convenient for users browsing or searching for data on individual species. Moreover, Kew's current Science Strategy sets out (pg. 27) an ambition to collaboratively deliver a 'Global Seed Information Facility'.

Kew launches two new MSc courses

As part of Kew's ambitious commitment to training the next generation of biodiversity scientists, we're expanding our postgraduate training offer with the launch of two new MSc courses. The MSc programmes offer the opportunity to study at Kew, with unrivalled access to our scientific collections, laboratory facilities, partnerships and landscapes.

Students can now choose between three MSc programmes at Kew in partnership with Queen Mary University of London or Royal Holloway, University of London:

Watch our short film: Study for an MSc at Kew

Further information on the MSc courses can be found:

Ten Golden Rules paper journals most downloaded in 2021

The paper "Ten golden rules for reforestation to optimize carbon sequestration, biodiversity recovery and livelihood benefits", published in the journal Global Change Biology last year was the journals most downloaded paper of 2021. Millennium Seed Bank staff and partners played a significant role in drafting this paper, including Alice Di Sacco, Kate Hardwick and Elinor Breman from RBG Kew, and Stephen Elliott from the Forest Restoration Research Unit, Chiang Mai University, Thailand. In addition, the paper formed part of the evidence base for the 'Kew Declaration on Reforestation for Biodiversity, Carbon Capture and Livelihoods' which was signed by over 3,000 global experts and citizens.




Issue 1: September 2021

This issue we hear about field work activities in Australia, Mexico, Pakistan and South Africa; we interviewed Janet Terry who retired from the Millennium Seed Bank in July after more than 30 years and we outline the current plant health requirements for shipping plant material to the UK.

Clusters of orange flowers on tree branches
Butea monosperma, Pakistan. (Photo: Amir Sultan).
Two purple flower heads emerging from rocks
Conophytum spp., South Africa. (Photo: J.A.S. James).
Several purple flowers on a branch with opposite pinnate leaves
Guaiacum coulterii, Mexico. (Photo: Jesús Sánchez).
A row of orchid seedlings with corresponding identification tags ready for translocation
Pterostylis psammophila, Australia. (Photo: Dr Jenny Guerin).

Pakistan is rated as ‘comparatively’ forest-poor by the FAO (Food and Agriculture Organisation of the United Nations), with only 0.05 ha of forest per capita compared with a global average of 1.0 ha in 2019 (FAO, 2002). In addition, the severe impacts of a changing climate, the high rate of population growth (1.95%) and the subsequent pressure on land, together with a high demand for forest products, is placing additional strains on the country’s forest cover.

In rural areas, wood is an important commodity for fuel and timber, which is driving forest clearance at an alarming rate (90% of households are reliant on wood for fuel). And additional pressures exist due to severe overgrazing and the conversion of forests for agriculture.

IUCN conservation assessments show that many trees used for their medicinal properties in Pakistan are threatened with extinction, such as the Critically Endangered Commiphora wightii (Mukul myrrh tree), while Taxus wallichiana (Himalayan yew) and Rhamnella gilgitica are classified as Endangered and Vulnerable respectively. Much research still needs to be carried out on other species to lift them from the status of Data Deficient, such as Cercis griffithii (Afghan rebud), Malus chitralensis (Chitral crab apple) and Prunus bokharienis.

Dr Shakeel Ahmad Jatoi and Dr Amir Sultan filling out paper forms in the field
Dr. Shakeel Ahmad Jatoi, Principal Scientific Officer, Bio-Resources Conservation Institute, left, and Dr. Amir Sultan, Programme Leader, Principal Scientific Officer, National Herbarium of Pakistan, record herbarium specimen data during a seed collecting trip to Chitral in June 2021. (Photo: Syed Bilal Hussain Shah).

A two-year project funded by the Garfield Weston Foundation in collaboration with the Pakistan Bio-resources Conservation Institute (BCI) is aiming to collect the seeds of at least 70 rare, threatened and over-exploited useful tree and shrub species in Pakistan. This project (part of the Global Tree Seed Bank Programme) follows on from the successful partnership between RBG Kew and BCI in the Medicinal and Aromatic Plants of Pakistan Project – MAPs (2019) and the Adapting Agriculture to Climate Change (Crop Wild Relative - CWR) project (2016-2018). Project funding has purchased essential equipment needed to increase the capacity of BCI staff to conserve seed collections, such as a seed aspirator, an incubator-drier, a freezer, and tree seed collecting equipment. Additional funding secured from the Alan Lennox Boyd Memorial Trust was used to purchase reference books and herbarium supplies.

Despite delays to the project due to COVID-19, the team in Pakistan led by Dr. Sadar Uddin Siddiqui (Chief Scientific Officer/Curator of the National Genebank of Pakistan and Director of the Bio-resources Conservation Institute) has been able to collect seeds and herbarium specimens from plants belonging to 18 families, 29 genera and 31 species. Collections in the 2020/2021 season were made across a variety of habitat types and include rarities in Pakistan, such as Wrightia arborea (Woolly dyeing rosebay) from which the root, bark and leaves can be used in traditional medicine, as well as Butea monosperma (Flame of the forest) and the climate change-resilient, but underutilized, Sideroxylon mascatense. This plant is a small, but viciously spiny shrub, that produces a sweet edible purple fruit (a bit like a plum), which is believed to have the potential to be brought more widely into cultivation.

Hannan Majeed and Syed Bilal Hussain Shah using a pole pruner to collect seeds from the top of a tree
Hannan Majeed, right, and Syed Bilal Hussain Shah, collect seeds of Flacourtia indica in Shahdara, Islamabad Capital Territory, as part of the medicinal trees and shrubs project (Photo: Sadar Siddiqui).
Two orange flowers
Flowers of Butea monosperma (Flame of the forest). (Photo: Amir Sultan).
Clusters of orange flowers on tree branches
Butea monosperma (Flame of the forest) in full bloom in Rawalpindi District, Pakistan, in April 2021. (Photo: Amir Sultan).
References
  • FAO (2002) An overview of forest products statistics in South and Southeast Asia. Eds. Ma, Q. & Broadhead, J.S.. EC-FAO Partnership Programme.

The Succulent Karoo biome stretches along the west coast of South Africa, spanning an area of approximately 111,000 km2 between southern Namibia and the uplands of the Western Cape Province. It is home to 5000 vascular plant species, 40% of which are endemic. The world’s richest succulent flora occurs here. This diversity is remarkable for a region that receives 100 to 200 mm of rain annually (Mucina et al. 2006). In addition to this limited winter rainfall, the plants depend on the contributions of fog, dew and water vapour to meet their water requirements (Matimati et al. 2012).

Landscape photograph of rocky slopes and distant mountains with little vegetation
The Northern Cape landscape. (Photo: J.A.S. James).

In the past two years, an unprecedented upsurge in wild plant poaching from the Succulent Karoo and the adjacent Nama Karoo biomes has escalated the need for conservation in these regions. The partnership between RBG Kew’s Millennium Seed Bank (MSB) and the South African National Biodiversity Institute (SANBI) has achieved over 7500 seed collections of native South African species to date. However, the species of the Succulent Karoo are relatively under-collected due to the high endemism of this biome and the remote locations of populations, which make accessibility for seed collections difficult. Thus, there are still many hundreds of rare and threatened plant species yet to be secured by seed banking.

The recent poaching increase is the result of a new interest in caudiciform and dwarf succulent species on foreign markets. Since mid-2019 there has been an increase in the number of poacher arrests and confiscation of wild plant material, which is brought to SANBI for identification, expert statements, and curation. In 2019 SANBI received on average 400 confiscated plants per week, which rose dramatically in 2021, to over 1500 plants per week. The genus Conophytum (Aizoaceae) has been especially targeted by poachers, while other confiscated genera include Anacampseros (Anacampserotaceae), Lithops (Aizoaceae) and Portulacaria (Portulacaceae). Many of these succulent species are already threatened and in some cases are critical habitat species, meaning that they only occur in an area less than 10 km2. This is the case for 48 Conophytum species, which occur as single populations, often on one ridge of a single mountain. Rare populations such as these are easily driven to extinction through over harvesting and several Conophytum species may already be extinct as a direct result of recent poaching.

Poaching compounds the pressure on these species from climate change. Models based on the key climatic and geological variables that influence Conophytum distribution predict a severe contraction of suitable habitat for most of the genus, increasing their extinction risk (Young et al. 2016).

In efforts to save these species from extinction in the wild, MSB South Africa has joined with the Custodians of Rare and Endangered Wildflowers (CREW), a SANBI programme that surveys and documents plant taxa of conservation concern, providing data essential for updating the IUCN Red List. A new MSB Karoo biome extension project is also currently being developed to respond to this conservation crisis.

Group image of 9 people in front of a dry mountainous landscape
Team photo: Maria Kotze, Nomndeni Nkosi, Jacqui James, Ruby Davies, Pieter van Wyk, Leandra Knoetze, Volenti van der Westhuizen, Adam Harrower and Gavin Links. (Photo: A.D. Harrower).

There is now a race to bank the seeds before populations are decimated by poaching. Earlier this year MSB South Africa joined CREW on a field trip to assess and bank seeds of the species currently under the most poaching pressure. It was incredible to see the effort that goes into succulent poaching. To find them, the team had to scramble up steep rocky mountainsides under the blazing sun in the dry expanse of the Northern Cape, our vehicles becoming specks in the distance. The dry remains of succulents that had succumbed to the severe drought in recent years were scattered across the landscape, a reminder of the delicate balance of this region and the losses that may be brought about by climate change.

A succulent plant on cracked ground with several seed capsules.
Aizoaceae: Cheiridopsis robusta. (Photo: J.A.S. James).
Two purple flower heads emerging from rocks
Conophytum growing on a cliff face. (Photo: J.A.S. James).

Species of the genus Conophytum grow as single bodies (smaller than your thumb) or in small dense clusters that camouflage well with their rocky surroundings. They often grow tucked into rock crevices and (in some cases) are only present on impossible to reach cliff faces, which may indicate that the more accessible plants have already been poached. In two cases, poachers were at the sites only a few hours before us. It was a privilege to see these species in their natural habitats, but tragic to know that if the poaching continues, we might be some of the last. Yet, there is still hope for these species, in the seeds of 11 species collected on that trip and others that will soon be stored in the vaults of the Millennium Seed Bank, protecting these species for the future.

References
  • Matimati, I., Musil, C.F., Raitt, L. & February, E. (2012) Non rainfall moisture interception by dwarf succulents and their relative abundance in an inland arid South African ecosystem. Ecohydrology. 6: 818-825. https://doi.org/10.1002/eco.1304.
  • Mucina, L., Jürgens, N., Le Roux, A., Rutherford, M.C., Schmiedel, U., Esler, K., Powrie, L.W., Desmet, P.G. & Milton, S.J. (2006) Succulent Karoo Biome. In: Mucina, L., Rutherford, M.C. (Eds.) The Vegetation of South Africa, Lesotho and Swaziland. Strelitzia 19. South African National Biodiversity Institute, Pretoria. pp. 220-299.
  • Young, A.J., Guo, D., Desmet, P.G. & Midgley, G.F. (2016) Biodiversity and climate change: Risks to dwarf succulents in Southern Africa. Journal of Arid Environments. 129: 16-24. https://doi.org/10.1016/j.jaridenv.2016.02.005.

Bridging the gap between the US Rocky Mountains and the Mexican Sierra Madre is a complex of 50 mountain ranges known as the Madrean ‘Sky islands’ Archipelago. With a diversity of habitats from desert shrubland to pine-oak woodlands, arranged across varied topography, the region is a priority for research and conservation, but botanical collections are under-represented from the Mexican territory (Deyo et al. 2013) and climate change is projected to erode and fragment the montane habitats (Yanahan & Moore 2018) before they have been fully studied.

Through the MSB Partnership, the Seed Bank of Facultad de Estudios Superiores Iztacala Universidad Autónoma de Mexico (FES-I UNAM) and RBG Kew have cooperated since August 2018 with the Herbarium of the University of Sonora (USON), in the important effort to conserve the Sonoran flora through seed banking. The aim was to diversify the collections of the seed banks involved and support their research studies, by collecting at least 150 accessions of native species of interest. Using USON in Hermosillo as the main base, field work was concentrated in 17 municipalities of Sonora State, as illustrated on the map in Fig. 1.

A map of Sonora state highlighting the municipalities of Agua Prieta, Alamos, Caborca, Cucurpe, General Plutarco Elías Calles, Guaymas, Hermosillo, Huásabas, La Colorada, Mazatán, Nácori Chico, Pitiquito, Puerto Peñasco, Santa Cruz, Suaqui Grande, TEpache and Ures.
Figure 1. Municipalities visited in Sonora State. (Map: FES-I UNAM).

During the 2018 and 2019 collection season, teams comprising staff from Kew, UNAM, and USON worked together in the field. The work was intense, the months of June, August and October of 2018 were the best for seed collecting. To maintain safety in these border regions the team were careful to seek cooperation with landowners and other biologists in the field. Although the 2020 health emergency forced many activities to stop, fortunately a team of local collectors located in Guaymas, led by Pablo Carrillo (Fig. 2), worked very hard despite the restrictions and managed to collect many species of interest.

Pablo Carrillo holding a pole pruner to collect seeds from the top of a tree he is standing next to
Figure 2. Biol. Pablo Carrillo collecting Parkinsonia microphylla in the town of La Pintada. (Photo: Pablo Carrillo).

Through the efforts of the teams, we have just finished the project with even better than expected results. In total, 244 accessions representing 44 botanical families, 147 genera and 194 species were collected and conserved, of which 49 are endemic species. Three species are included in some category of risk in accordance with the Official Mexican Standard NOM-059 SEMARNAT 2010, 14 are in Appendix II of CITES and 17 are in the Least Concern (LC) category of the IUCN.

All the species collected have biological, ecological, or economic importance, however the following species are especially emblematic:

  • The Guayacán, Guaiacum coulterii A. Gray, a native tree of the thorny scrub of Sonora, has medicinal properties and its wood is useful for construction as well as being very beautiful as an ornamental plant and a source of food for different species of pollinators. It is classified as threatened in Mexico and vulnerable internationally (Fig. 3 & 4).
    Several purple flowers on a branch with opposite pinnate leaves
    Figure 3. Flower of Guayacán, Guaiacum coulterii. (Photo: Jesús Sánchez).
    Three small red oval fruits of Guaiacum coulterii hanging on the end of a branch
    Figure 4. Fruit of Guayacán, Guaiacum coulterii. (Photo: Jesús Sánchez).
  • The Cabeza de viejo (‘Old Man's Head’), Lophocereus schottii (Engelm.) Britton & Rose, is a columnar cactus which has medicinal properties, found in Sonora’s thorny scrub and dunes. It is classified as subject to special protection in Mexico.
  • The Saguaro, Carnegiea gigantea (Engelm.) Britton & Rose, is perhaps one of the most appreciated species for its beauty in Arizona and Sonora. It has threatened status in Mexico.
  • A Nopal, an Opuntia spp., collected in the Sierra de Mazatán, is probably a new species for science that is being described by specialists.

In conclusion, the state of Sonora is, without a doubt, a place rich in plant diversity that should continue to be studied and conserved. Much remains to be discovered, collected and conserved in the most remote and inhospitable areas that are unfortunately not very easy to access.

Thanks to this project, valuable information has been created on the flora of the sky islands within Sonora, and knowledge has been developed on how security can be maintained within the area, also on the bi-national distribution of species that are threatened in the United States and little known from Mexico. We acknowledge generous support of John and Catherine Emberson to the Kew Foundation to enable this project to happen.

References

Over the last month, staff at the South Australian Seed Conservation Centre have been undertaking several orchid translocations. We translocated 116 plants of Sandhill Greenhood (Pterostylis arenicola) to Torrens Island, with help from Coastal Officer Darren Kennedy to measure and tag plants. Darren will water and help monitor the plants. We also translocated some Bayonet Spider-orchids (Caladenia gladiolata) to a private heritage agreement near Scott Creek Conservation Park (CP). In addition, Jenny and Thai translocated the Two Bristle Greenhood (Pterostylis psammophila) to exclosures in Sandy Creek CP last week, with the help of Kym Smith from the Friends group who helped record, tag and plant the greenhoods in the exclosures. Kym will help monitor this translocated population in the future.

Thai and Kym kneeling down plant orchids
Thai Te and Kym Smith measuring, tagging and planting the endangered Pterostylis psammophila at Sandy Creek Conservation Park. (Photo: Dr Jenny Guerin).
A row of orchid seedlings with corresponding identification tags ready for translocation
Pterostylis psammophila plants propagated using symbiotic in-vitro methods ready for translocation. (Photo: Dr Jenny Guerin).

In the lab we’ve also isolated fungus for a number of threatened orchid species lately for some symbiotic in-vitro propagation work later this year. We’ve been trialling some methods for some difficult species so we were recently delighted (and relieved) to see some success for black-beaked duck-orchid (Caleana disjuncta). We’re doing further research on the Copper beard-orchid (Calochilus cupreus) at the moment for ‘Green Adelaide’ so we’re hoping for similar success before we take off into the wilds again. The world count got up to nearly 40 plants last year so we’re hoping to augment this with propagated plants…and find more populations.

Magnified image showing the tear drop shaped seed of Caleana disjuncta with fungal hyphae protruding
Black-beak duck orchid (Caleana disjuncta) protocorm on oatmeal agar media. (Photo: Dan Duaval, SASCC).

On the 27th August, we put on a display at Adelaide Botanic Garden of some of the endangered orchids (12+ species) we’re currently propagating for the ‘Back from the Brink’ project. This was a good opportunity for landholders, volunteers, partners and sponsors to see some of these interesting orchids in flower.

Kolkhuri jonjoli, Staphylea colchica Steven

Ian Willey (RBG Kew)

Staphylea colchica is native to the Caucasus region where it is known commonly as kolkhuri jonjoli (კოლხური ჯონჯოლი Georg.), Colchis Bladdernut, Caucasian Bladdernut, Jonjol or the Jonjoli tree. Staphylea colchica is a deciduous shrub or small tree growing up to 4 metres in height with three or five ovate leaflets with finely toothed margins. In spring its small, white, fragrant bell-shaped flowers are borne on racemes and develop into greenish-white bladder-like papery fruits containing orange seeds (hence its name “bladdernut tree” in English). The flowering shoots with buds of ‘Jonjoli trees’ are edible and are pickled by fermenting in brine to produce Jonjoli, a revered delicacy in the Caucasus, and in Georgia are a common feature of the traditional feast “Supra”.

People from across the Caucasus region will have eaten Jonjoli, and an export market exists in countries with a significant Caucasian diaspora. S. colchica’s cultural value is both a blessing and a curse. Jonjoli occupies a special place in the region’s cultural identity with the tree recognisable and cherished by many. However, wild populations are threatened by the over collecting of reproductive parts for culinary purposes. Harvesting the flowering shoots and buds limits the plants’ ability to reproduce from seed, further decreasing the viability of source populations. Additionally, S. colchica distribution is severely fragmented across its native range, and the number of individual plants per population is low. A preliminary assessment in 2005 reports that distribution in the Caucasus has reduced by 50%, but a full global IUCN Red List assessment remains incomplete, and more field data is needed to establish a reliable baseline. Taxonomic studies are needed to validate species status and more research is needed to understand threats to the species and the true rate of decline.

Two ex situ seed collections of S. colchica are banked in Georgia and duplicated to the MSB. These collections are a key component of efforts to protect this species and seeds can be made available for use dependant on prior consent from partner institutions. However, S. colchica requires greater protection in situ. Overharvesting and land-use change are very real threats but by promoting sustainable harvesting and engaging communities, as was done in the Darwin Initiative-funded Enhancing rural Caucasian livelihoods through fruit and nut conservation, this culturally significant plant can be protected.

More information on other species conserved through the project can be found in the project booklet (pdf).

A cluster of white flowers and buds
The flowers and buds of the Jonjoli tree. (Photo: National Botanic Garden, Georgia).
A group of people standing in a circle listening to a member of the British Georgian Society in front of a Jonjoli tree
The Georgian Ambassador to the UK, Her Excellency Ms Sophie Katsarava, with the British Georgian Society presenting the cultural importance of Jonjoli whilst standing in front of Staphylea colchica at Kew Gardens, July 2020 (Photo: I. Willey).

A fond farewell to Janet Terry

Three people stood holding a framed certificate infront of the Svalbard Global Seed Vault.
Janet (centre) as part of the MSB team delivering seeds to the Svalbard Global Seed Vault on behalf of The Prince of Wales. (Photo: Ahmed Amri).
Janet holding a small glass vial containing a seed
Janet holding the billionth seed to be banked at the MSB. (Photo: Paul Little).

Circa 1984, when Janet began a Saturday job working in the restaurant at Wakehurst, she could not have envisaged the 3-decade long career which would follow, when she would go on to travel the world and meet, train, and collect with “so many of our wonderful partners” and “see so many strange and unique seeds and know we have them safely stored”.

When an opportunity arose with the International Board for Plant Genetic Resources (IBPGR, now The Crop Trust) which was running a seed handling unit at Wakehurst, Janet applied for the job as “it sounded interesting!”. Teams went out in the field (predominantly Africa) to collect cultivars and specimens of all the major crops. These got sent back to Janet, who had to sort and distribute them to the crop gene banks which were working on them. In 1990 a seed processing job came up with Kew and Janet moved to work for Kew. Work started on setting standards for collecting and storing seeds. As Janet says, these standards remain largely unchanged today – “keep it dry, keep it cool and it will live longer”.


We asked Janet to reflect on her incredible journey:


What will you miss most about working at the MSB? The people – definitely! Especially the contact with counterparts in equivalent positions around the world, who we can share ideas and experiences with.

What's the favourite species you have handled? The grapple - Harpagophytum. We collected them in Botswana. They were quite challenging to collect and even more so to clean!

What's your first memory of working here? That the Seedbank was in the Mansion – Lady Price’s bedroom was the lab and Sir Henry Price’s bathroom was the x-ray room. The stark contrast of working in an Elizabethan mansion whilst concentrating on cutting edge work! There was a large computer which took up a whole room in the Orchard building. In those early days the field data sheets were loaded onto this huge computer, so from the outset the data has been stored digitally.

What advice would you give to somebody about how to clean a collection? Slowly and carefully!

How do you see seed processing changing in the future? Funding for ‘business as usual’ processing is difficult. People love to fund new and exciting projects, but we need to be able to sustain the day-to-day processing, otherwise we may end up without processing roles and just dry and store seed at -20°C. Without viability testing, we cannot distribute the seed. This highly skilled work by practical applied scientists needs to be supported. I wish their skills were valued more highly. It’s also learning from others – the partnership, collaboration and knowledge that is the MSB’s strength.

If you hadn't worked here, what else might you have liked to do? When I worked for IBPGR, I went to a conference at their headquarters in Rome and a network contact from Indonesia offered me a job working with Orangutans. I was very tempted - things could have been very different!

What are you planning to do in your retirement? Nothing at first - that’s the point! Then I’ll probably get a part time job, working outdoors.

If you could be one plant, what would it be and why? A Welwitschia, as it’s slow growing and they live for an awfully long time. I would sit and watch the world go by!

A close up image of a woody seed pod with several stalks protuding with hooks on
A Harpagophytum fruit. (Photo: Elly Vaes, RBG Kew).
A single Welwitschia plant in the desert
Welwitschia mirabilis. (Photo: Andrew McRobb).

Somehow, we can’t imagine Janet letting the grass grow under her feet for very long! Thank you to Janet for sharing her thoughts and memories with us.

What are the new requirements for sending plant material to the MSB?

Due to the UK leaving the European Union, there have been changes to Plant Health rules for shipping plant material to the UK.

All seed collections coming into the MSB from outside the UK now need to come with a phytosanitary certificate issued by the relevant authority in the exporting country. A phytosanitary certificate is an official document declaring that plants and plant material are free from pests and diseases. If seed collections arrive with this documentation, then none of the collections are classed as quarantine and the seeds and plant material will be processed as normal. This applies to wild collected and commercial seeds and plant material. The processing and long-term storage of the collections will be no different.

If a phytosanitary certificate cannot be obtained, then material will need to come in with a Letter of Authority (LoA) from the UK Plant Health Service, which can be obtained from the MSBP Conservation Coordinator for your region. However, any material coming in under the LoA will be treated as 'quarantine' upon arrival and restrictions will be placed on the future use of these collections.

As usual, before seed collections are dispatched to the MSB, species lists should be sent to the Seed Collections Manager who will check the list for any prohibited species and CITES-listed species (the Convention on International Trade in Endangered Species of Wild Fauna and Flora).

Copies of all documents should be attached to the outside and also put inside the shipping box. Please also send the airwaybill (AWB) number and copies of the documents to the Seed Collections Manager, so the consignment can be tracked.

A desk with a row of paperwork, a row of seed packets and a set of herbarium specimens partially wrapped in newspaper on
Preparing the paperwork, seeds and herbarium specimens for shipment. (Photo: RBG Kew).

Recent publications from across the MSBP:

Updates to the field data form

We are currently in the process of updating the field data form to improve its compatibility with the data required for IUCN threat assessments. We expect the update to be ready before the end of December.