Search

Conservation Translocation (CT), which includes reintroduction, reinforcement and introduction, is gaining momentum, responding to serious population decline in many orchids. Orchid conservation underpinned by a greater emphasis on understanding biotic and abiotic factors of habitats is critical for successful recovery and restoration programmes. Anacamptis morio, commonly known as green-winged orchid (GWO), is a terrestrial orchid found throughout Europe, but populations in England rapidly declined in its native range in the last several decades. The current study explored the relationship between soil abiotic characteristics and the community composition of key fungal groups. Wild sites in Essex and Cumbria in England, where successful colonies are currently present, were compared to potential sites for CT in Cumbria. The Cumbrian sites managed by cattle grazing include the wild site and three potential CT sites, with two of them hosting no GWO plants. The Essex site, fOxley Meadow, where no cattle is used for grassland management, hosts the largest population of GWO in England. The aim of this study was to understand whether the community composition of fungi and soil characteristics of the potential CT sites in Cumbria are nearly compatible with that of Oxley Meadow. Oxley Meadow, with around 65,000 plants, stands out as a unique habitat compared to all Cumbria sites, as it showed low organic content. Nitrate and phosphate content were smaller for Oxley Meadow compared to other Cumbrian sites. The proportion of Basidiomycota fungi was greater in Oxley Meadow compared to all Cumbrian sites where Ascomycota dominated. The abundant fungal group found in Oxley Meadow was Agaricales. From Agariclaes, Hygrophoraceae or waxcaps fungi are considered an indicator group of fungi and were the most abundant group in Oxley Meadow. They have a negative correlation with targeted key fungal groups and abiotic parameters. More in-depth assessments using additional primers are essential to better understand the fungal diversity and how this diversity translates to the resilience of orchid habitats. This preliminary study points to future studies to assess whether sites are near-compatible to the wild site where large colonies are present using additional primers collected over different time scales. [ABSTRACT FROM AUTHOR]

The yellow early marsh orchid (Dactylorhiza incarnata ssp. ochroleuca) is a critically endangered terrestrial orchid in Britain. Previous attempts to translocate symbiotic seedlings to a site near the last remaining wild site demonstrated some success, with a 10% survival rate despite adverse weather conditions over a two-year period. However, to facilitate future reintroduction efforts or conservation translocations, a more comprehensive understanding of the fungal microbiome and abiotic soil characteristics at the final remaining wild site is required. Obtaining comprehensive information on both the fungal community and soil nutrient composition from wild sites has significant benefits and may prove critical for the success of future conservation translocations involving threatened orchids. This preliminary study, conducted at the last remaining wild site, revealed a significant correlation between the relative abundance of the orchid mycorrhizal fungal order Cantharellales and the concentrations of nitrate and phosphate in the soil. Another orchid mycorrhizal fungal group, Sebacinales, was found to be distributed extensively throughout the site. The composition of fungal communities across the entire site, orchid-hosting and non-orchid-hosting soils is discussed in relation to reinforcing the current population and preventing the extinction of this orchid. [ABSTRACT FROM AUTHOR]

The split between conservation science and real‐world application is an ongoing issue despite several calls for unification. Researchers are empowered to partially bridge the research‐implementation gap by making their findings more accessible. Cypripedium calceolus is the most recognizable orchid of the European flora, and is currently facing habitat change and fragmentation, in addition to threats from collectors and illegal traders. Although several studies have focused on the ecological and genetic features of the species, a comprehensive account of how such aspects can be translated into concrete conservation recommendations is still missing. In this study, we describe microsatellite genetic variation in 188 individuals from different Italian populations of C. calceolus. Our results indicate the need for immediate conservation action for the most isolated populations in the Central Apennines and north‐western Italy. Although our genetic findings are specific to the Italian populations, our aim is to review ecological and population genetic aspects in C. calceolus and their implications for conservation against the existing threats. Therefore, our detailed guidelines for translocation, habitat management and post‐translocation monitoring can be used to inform conservation strategies in threatened populations of C. calceolus across its range. [ABSTRACT FROM AUTHOR]

Orchid mycorrhizal fungi (OMF) are critical for seed germination and maintaining natural populations of orchids, yet the degree of specificity of most orchids to their mycorrhizal associates remains unknown. Many orchids are at risk of extinction, whether generalists or specialists, but orchid species of narrow fungal specificity are arguably under increased threat due to their requirement for specific fungal symbionts. This study characterises the fungi associated with Aerangis ellisii, a lithophytic orchid from a site in the Central Highlands of Madagascar. Culturable OMF isolated from spontaneous protocorms of this species from the wild were used for seed germination. In vitro germination and seedling development of A. ellisii were achieved with fungi derived from A. ellisii and an isolate from a different Aerangis species 30 km away. The significance of these findings and their importance to conservation strategies for this species and other Aerangis spp. is discussed. These results have important implications for the conservation of A. ellisii populations in Madagascar. [ABSTRACT FROM AUTHOR]

Cephalanthera rubra (L.) Rich., Red Helleborine, is a widespread orchid in Europe but known only from three very small populations in England. These populations are in decline with no natural seed setting for more than a decade. The species may become extinct in the UK soon unless viable strategies are in place for ex situ conservation, especially the use of symbiotic propagation. Because of the fragile nature of the populations in England mycorrhizal fungal diversity study is not feasible. Therefore, to understand the factors needed for healthy Red Helleborine populations, soil characteristics and diversity of culturable root-derived fungi of the populations from a small area in the Loire Valley in France were studied. Themain objectives of the study were: (1)Which culturable mycorrhizal fungi associated with C. rubra roots and (2) To what extent is variation in fungal communities related to variation in soil characteristics? Here, we report a significant difference in diversity of culturable mycorrhizal and non-mycorrhizal fungi depending on soil pH and phosphorus content. Mycorrhizal associations were favoured by plants in locations with low soil nutrient availability and comparatively higher pH. Our study shows that mycorrhizal fungi, both ecto and endo, can be cultured from roots of plants at different maturity stages. [ABSTRACT FROM AUTHOR]

Background: Tropical orchids need more study with respect to their mycorrhizal associations. For researchers in distant countries who aspire to study these orchids augmenting their conservation, the great distances involved, coupled with limited funds, pose formidable challenges. These challenges are sometimes exacerbated by political unrest, delays in securing permits, unexpected hardships, and the risk that the biological samples collected (e.g., roots harboring mycorrhizal fungi) will not survive long-distance transport. Results: We describe a protocol for the collection and transport of root samples from Madagascar orchids to labs in the United Kingdom (Kew) and the United States (Illinois) where Rhizoctonia-like fungi were subsequently isolated. Three separate trips were made spanning 4 years (2012-2015), with emphasis on the collection of roots from epiphytic, lithophytic, and terrestrial orchids inhabiting the Itremo Massif of the Central Highlands. Collectively, the trips to Madagascar resulted in the isolation of all major groups of Rhizoctonia-like fungi ( Ceratobasidium, Tulasnella, Sebacina) from all three orchid growth forms (terrestrials, epiphytes and lithophytes). Sampling of terrestrial and epiphytes during the rainy season (January) yielded best results. Conclusions: Our study demonstrates that peloton-forming fungi in root samples can retain viability up to 3 weeks after collection. [ABSTRACT FROM AUTHOR]

Background: Lack of phenological information and efficient collecting methods are considered impediments for orchid seed collecting. This leads to opportunistic collecting as part of general seed collecting schedules that may last few weeks especially in remote areas. The study explored the feasibility of collecting near mature and immature seeds to support conservation action plans. Mature, near mature and immature seeds of orchids were collected from the wild in the Central Highlands of Madagascar (CHM). Seed capsules were collected in sterile culture medium in the wild, to prevent deterioration of seeds inside the capsule after collecting, later to be cultured under laboratory conditions. Results: Seed capsules collected by the in vitro collecting (IVC) method were kept in very good condition for up to 4 weeks before germination under in vitro conditions. Significantly faster and higher germination rate ( p < 0.001) than mature seeds was observed in a number of taxa collected during a 3 year-long study. In some taxa even immature seeds, with no apparent sign of testa covering the embryo, germinated following IVC where mature seeds failed to germinate. Conclusions: We propose that IVC method has potential to complement conventional seed collecting by increasing the germplasm that can be used in integrated conservation action plans. Improvements can be made in developing collections for taxa from biodiversity hotspots and remote areas where collecting requires considerable resources. This method can further be used on a wider selection of plants from different geographic areas and on embryo rescue programmes for economically important plants. [ABSTRACT FROM AUTHOR]

Habitat fragmentation and invasive alien species contribute to genetic bottlenecks and the threat of extinction of many endemic species of Mauritius, part of the Madagascan and Indian Ocean biodiversity hotspot. The genus Cylindrocline has two species, C. commersonii (critically endangered) and C. lorencei (extinct in the wild). The last living specimens of C. lorencei disappeared in the wild after the recorded collecting of seeds in 1982 by Conservatoire Botanique National de Brest (CBN Brest). Embryo rescue was used as a method to germinate these seeds and the seedlings raised this way were shared by CBN Brest with Royal Botanic Gardens Kew (RBG Kew) as part of an exchange programme in 2001. Mature plants both at CBN Brest and RBG Kew stopped producing viable seeds and this has made the long-term conservation of C. lorencei even more difficult. Seeds of C. commersonii collected from the wild in 2010 have a very low viability while ex situ grown C. commersonii produce non-viable seeds. Molecular studies conducted in C. lorencei using amplified fragment length polymorphisms (AFLP) showed no genetic variability among remaining individuals. Two samples of C. commersonii showed a very small amount of genetic variability. The variability between the two species was well within the limits commonly found within species or between closely related species and the long-term conservation of the genus requires a radical (to a degree) approach to avoid its extinction. The importance of novel approaches for restoration and long-term conservation are discussed. [ABSTRACT FROM AUTHOR]

The Orchid flora of Madagascar is one of the most diverse with nearly 1000 orchid taxa, of which about 90 % are endemic to this biodiversity hotspot. The Itremo Massif in the Central Highlands of Madagascar with a Highland Subtropical climate range encompasses montane grassland, igneous and metamorphic rock outcrops, and gallery and tapia forests. Our study focused on identifying culturable mycorrhizae from epiphytic, lithophytic, and terrestrial orchid taxa to understand their diversity and density in a spatial matrix that is within the protected areas. We have collected both juvenile and mature roots from 41 orchid taxa for isolating their orchid mycorrhizal fungi (OMF), and to culture, identify, and store in liquid nitrogen for future studies. Twelve operational taxonomic units (OTUs), of three known orchid mycorrhizal genera, were recognized by analysis of internal transcribed spacer (ITS) sequences of 85 isolates, and, by comparing with GenBank database entries, each OTU was shown to have closely related fungi that were also found as orchid associates. Orchid and fungal diversity were greater in gallery forests and open grasslands, which is very significant for future studies and orchid conservation. As far as we know, this is the first ever report of detailed identification of mycorrhizal fungi from Madagascar. This study will help start to develop a programme for identifying fungal symbionts from this unique biodiversity hotspot, which is undergoing rapid ecosystem damage and species loss. The diversity of culturable fungal associates, their density, and distribution within the Itremo orchid hotspot areas will be discussed. [ABSTRACT FROM AUTHOR]

Last seen in 1958, the Ascension Island endemic fern, Anogramma ascensionis, was listed as extinct on the 2003 IUCN Red List. However, a 2009 survey rediscovered four plants on Green Mountain. Spores were collected and cultured in vitro at the Royal Botanic Gardens, Kew, where a living collection of thousands of gametophytes and hundreds of sporophytes has been developed. To gain further insights into the biology of this species and the potential implications of in vitro multiplication for conservation purposes, samples were characterized from the karyological point of view. Chromosome analysis of root tips has confirmed that the species is tetraploid, and flow cytometry assessments have revealed that haploid gametophytes produce diploid sporophytes, which confirms natural fertilization. In addition, an rbcL sequence from A. ascensionis has been generated and compared with those published for other Anogramma spp., suggesting a close relationship with A. chaerophylla from Brazil. Further surveys of Green Mountain have reported the presence of 40 A. ascensionis sporophytes in total. Vegetation community analyses have suggested that the present population may be confined to suboptimal habitats. We therefore propose that, prior to the dramatic transformation of the vegetation on the island as a result of the invasion of alien species (particularly Adiantum spp.), A. ascensionis may have flourished in more humid and shaded parts of the mountain. A multidisciplinary approach involving in vitro culture, invasive species clearance and controlled translocation is discussed as the future roadmap for the conservation of this critically endangered fern. Our experiences have also highlighted lessons more broadly applicable to the conservation of extremely rare species elsewhere in the world, especially on remote island systems. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174, 461-477. [ABSTRACT FROM AUTHOR]

Plants provide medicine and pest control resources for millions of poor people world-wide. Widespread harvesting of medicinal and pesticidal plants puts pressure on natural populations, thus severely compromising their contribution to the income and well-being of traders and consumers. The development of in vitro propagation techniques appropriate for developing countries will provide a robust platform for effective propagation and cultivation of endangered plants. This review focuses on advances in the application of phytochemical and in vitro tools to identify and rapidly propagate medicinal and pesticidal plants. Problems of over-harvesting can be alleviated and ex situ cultivation in agroforestry systems can be facilitated through improving seed germination, in vitro cloning and the use of mycorrhizal fungi. We also present a case for effective use of phytochemical analyses for the accurate identification of elite materials from wild stands and validation of the desired quality in order to counter loss of efficacy in the long run through selection, propagation or ex situ management in agroforestry systems. Future prospects are discussed in the context of medicinal activity screening, sustainable propagation, on-farm planting, management and utilization. [ABSTRACT FROM AUTHOR]

The importance of in vitro tools to complement other ex situ methods for saving plants from extinction is more relevant than ever before. More than 50% of the world's plant species are endemic to the 34 global biodiversity hotspots (GBHs), each holding at least 1,500 endemic plant species. In addition, a large number of small islands hold a number of endemic species on the brink of extinction. Conservation support concentrating more on these hotspots and small islands would significantly reduce the loss of species that is currently occurring. In the majority of these cases, the resources are either locally scarce or difficult to access for in vitro conservation to support other ex situ measures. Most island countries are small, and their geographical position is a stumbling block to initiate active partnerships with other countries when they need to use in vitro tools to rescue plants that produce recalcitrant seeds/spores or propagate only by vegetative means. However, many biodiversity hotspot countries have facilities and expertise, and they concentrate on their own flora for in vitro conservation programmes. For decades, because of the grave threat these plants face, the Conservation Biotechnology Unit, previously known as the Micropropagation Unit, at Royal Botanic Gardens Kew (RBG Kew) has been at the forefront of assisting countries to save their valuable biodiversity through both in situ and ex situ methods. Approaches mentioned here highlight work on recalcitrant ferns from GBHs and small islands. Source materials from recalcitrant species, either spore or seed and in some cases vegetative material, need to be used immediately after collection for tangible results in vitro. This becomes more difficult when only a few plants or small populations are left in the wild. The task becomes harder when available material is small in quantity, and there is greater restriction on the use of available genetic diversity in the wild. This paper highlights the importance of proper collection measures, in vitro culture procedures and cryopreservation and methods for the integrated conservation of threatened ferns from both GBHs and small islands. The importance of international networking to achieve these conservation goals also will be discussed. [ABSTRACT FROM AUTHOR]

This special issue is dedicated to the in vitro tools and methods used to conserve the genetic diversity of rare and threatened plant species from around the world. Species that are on the brink of extinction because of the rapid loss of genetic diversity and habitat come mainly from resource-poor areas of the world and from global biodiversity hotspots and island countries. These species are unique because they are endemic, and only a few small populations or sometimes only a few individuals remain in the wild. Therefore, the challenges to support conservation by in vitro measures are many and varied. The editors of this invited issue solicited papers from experts from Asia, Africa, Europe, Australia, and North and South America. This compilation of articles describes the efforts in these diverse regions toward saving plants from extinction, and details the direct application of in vitro and cryopreservation methods. In addition, these contributions provide guidance on propagation of rare plants, including techniques for large-scale propagation, storage, and reintroduction. The in vitro techniques for conserving plant biodiversity include shoot apical or axillary-meristem-based micropropagation, somatic embryogenesis, cell culture technologies and embryo rescue techniques, as well as a range of in vitro cold storage and cryopreservation protocols, and they are discussed in depth in this issue. [ABSTRACT FROM AUTHOR]

Medusagyne oppositifolia is a critically endangered monotypic species with only a few trees left in the wild. Due to invasion and habitat clearance, genetic diversity of this species has been reduced to an alarmingly low level. Long-term in vitro cultures of M. oppositifolia were slow-growing and eventually lost the ability to multiply and root. Because of the lack of new seed and vegetative materials, efforts were made to rejuvenate the existing cultures. This was attempted by a series of steps involving serial pruning of the shoot tips and growing on a modified MS medium containing silver nitrate and thidiazuron. Rapid cycling clonal propagules were produced this way from cultured nodes. Isolated shoots were rooted on Florialite™, and rooted plants were weaned in a transition stage using a Nutrient Film Technique (NFT) hydroponic system before being completely weaned under glasshouse conditions. The importance of screening cultures for multiplication and rooting efficiency during long-term maintenance is discussed in the context of stress metabolite production and its influence on growth in culture. Factors contributing to rejuvenation of old cultures, application of unconventional supporting systems for rooting and application of the hydroponic system as a weaning tool are also discussed. [ABSTRACT FROM AUTHOR]

The latest IUCN statistics show that of over 12,000 plant species, 70% are threatened, 19% are critically endangered and 28 species are extinct in the wild. Target 8 of the Global Strategy for Plant Conservation (GSPC) highlights the importance of ex situ conservation of critically endangered plants. Long-term germplasm storage for species with recalcitrant seeds needs alternative measures. In vitro methods complement seed banking and other ex situ measures and are vital for long-term conservation. Conservation Biotechnology at RBG Kew is currently working on a number of rare and threatened recalcitrant species from biodiversity-rich areas of the world to develop good quality in vitro propagules for cryopreservation, recovery and restoration projects. The importance of successful in vitro propagation methods, transplantation technologies, cryopreservation and international networking for the integrated conservation of these species are discussed in detail. [ABSTRACT FROM AUTHOR]

Polyploids have significantly influenced angiosperm evolution. Understanding the genetic consequences of polyploidy is advanced by studies on synthetic allopolyploids that mimic natural species. In Nicotiana, Burk (1973) and Kostoff (1938) generated synthetic tobacco (N. tabacum) using the parents ♀N. sylvestris x ♂N. tomentosiformis. We previously reported rapid genetic changes in the Burk material. Kostoff's material has 24 chromosomes of N. sylvestris origin (S-genome), 24 of N. tomentosiformis origin (T-genome), and a large intergenomic translocation, but not an additive distribution of ribosomal DNA (rDNA) families as expected from the parental contribution. Our new synthetic tobacco lines TR1 and TR2 are chromosomally balanced with no intergenomic translocations and are either sterile or have highly reduced fertility, supporting the nuclear cytoplasmic hypothesis that allopolyploid fertility is enhanced by intergenomic translocations. Two plants of TR1 (TRI-A, TRI-B) have the expected number, structure, and chromosomal distribution of rDNA families, in contrast to Burk's and Kostoff's synthetic tobaccos and to synthetic polyploids of Arabidopsis. Perhaps allopolyploids must pass through meiosis before genetic changes involving rDNA become apparent, or the genetic changes may occur stochastically in different synthetic allopolyploids. The lack of fertility in the first generation of our synthetic tobacco lines may have uses in biopharmacy. [ABSTRACT FROM AUTHOR]

The yellow early marsh-orchid (Dactylorhiza incarnata subsp. ochroleuca) is critically endangered in the UK. Reintroduction of this threatened orchid to former haunts that have been restored is a long-term objective of this study. Identifying germination-specific mycorrhizal fungus lineages from closely related species is used as a method due to the extremely small number of plants left in the wild. A putative orchid mycorrhizal fungus of the family Tulasnellaceae, isolated from Dactylorhiza praetermissa, supported in vitro seed germination to produce reintroduction-ready seedlings. Reintroduced symbiotic seedlings survived over the winter months in the flooded reintroduction site (RS). The comparative soil analysis for key nutrients before reintroduction showed that phosphorus content in the RS is very low compared to the soil collected from the wild site (WS) where the last viable population exists. On the other hand, C:N ratio in the soil at the WS and RS were not significantly different. To our knowledge, this is the first-ever report on the reintroduction of symbiotic seedlings of a threatened orchid back to the wild in the UK. [ABSTRACT FROM AUTHOR]

About 90% of Cynorkis species are endemic to the biodiversity hotspot of Madagascar. This terrestrial habitat-specific genus received little study for fungal diversity to support conservation. We evaluated the diversity of culturable fungi of 11 species and soil characteristics from six sites spanning a >40 km radius in and along the region's inselbergs. Peloton-forming fungi were grown in vitro from root/protocorm slices and positively identified using DNA sequencing. The fungal diversity was then correlated with soil pH, NO3-N, P, and K. All species harbored either putative mycorrhizal associates in the Rhizoctonia complex or Hypocreales fungi. Tulasnella Operational Taxonomic Units (OTUs) were most prevalent in all soil types while Serendipita OTUs were found in species inhabiting granite/rock outcrops in moist soil (seepage areas). Most Cynorkis species were present in soil with low NO3-N and P levels with diversity of mycorrhizal fungi inversely correlated to NO3-N levels. Of the different orchid life stages sampled, only one species (Cynorkis fastigiata) yielded putative mycorrhizal fungi from juvenile stages. As diversity of mycorrhizal fungi of Cynorkis spp. was negatively correlated with NO3-N, and majority of the studied taxa were found in soils with low NO3-N and P contents, reintroduction studies must include analysis of N and P in detail. For the first time, we showed that the assemblage of culturable fungi in the roots of habitat-specific species of Cynorkis (Orchidaceae) are intimately tied to specific soil characteristics. [ABSTRACT FROM AUTHOR]

The inselbergs of the Central Highlands of Madagascar are one of many 'micro-hotspots' of biodiversity on the island, particularly for Orchidaceae. In this region are several genera that have a large number of endemic species that are in serious decline or edging towards extinction. Studies relating to diversity of orchids and their fungal partners (both mycorrhizal and non-mycorrhizal root associates) deserve more attention, as climate change and human induced decline in resilience of species in the wild is at an all-time high. Identification of mycorrhizal fungi (MF) via conventional seed baited-protocorms has limitations for large scale studies and its application for time-bound conservation projects. The paper describes the value of understanding fungal diversity in the roots of orchids at different stages of maturity. The first part of the study was a preliminary investigation mainly to identify culturable Rhizoctonia endophytes, and the second part looked at all life forms of available taxa together with associated soil characteristics. We isolated and identified 19 putative MF from 18 of the 50 taxa spread over an area of 250 sq. km, covering three life forms, growth phases of the orchid taxa, and habitat types. In the rest of the taxa, we were unable to detect any putative MF, but had varying numbers of non-mycorrhizal endophytes. We also found that diversity of putative MF was higher in plants from soils with the lowest P levels recorded. Putative mycorrhizal OTUs were predominantly from the Tulasnella lineage, followed by Ceratobasidium and Serendipita. Within a small subset of samples, a difference in colonised endophytes depending on the collection season was observed. In vitro germination studies using 10 OTUs of mycorrhizal fungi in 14 orchid species showed mostly generalist associations. When orchid seed and fungal sources were studied irrespective of habitat, life form, and distance from each other (orchid seed and fungal source), compatibility for symbiotic seed germination was observed in most cases. Issues with the identification of compatible MF and symbiotic system of seed germination are discussed. [ABSTRACT FROM AUTHOR]

Background: Orchids are under threat from human activities and climate change, with populations limited to small geographic hotspots. This makes them ideal candidates for ex situ conservation. Orchid seeds are desiccation tolerant, but often have poor longevity in seed banks and cryopreservation of orchid protocorms is complex and expensive. Therefore, simple methods for large-scale storage programs are essential to store orchid seeds of different life forms. Seeds of five species representing epiphytic, lithophytic and terrestrial orchids from the Central Highlands of Madagascar were studied to find a simple and effective system of cryopreservation. The use of a vitrification solution prior to cryopreservation to improve survival was investigated, as well as the use of symbiotic and asymbiotic germination media to maximise germination after cryopreservation. Using the filter paper packet method, dried seeds were stored in vapour phase above liquid nitrogen and recovered after thawing with both symbiotic and asymbiotic media.Results: The study revealed that cryoprotection is not essential for the species in this study, which represented a range of lifeforms. Vitrification generally led to a decrease in germination post cryopreservation. The use of a symbiotic germination medium post cryopreservation was found to be successful in the species in which it was tested. However, the use of an asymbiotic medium was successful for all the species in this study.Conclusions: Vitrification was not essential for the species in this study as the orchid seeds were already ultralow temperature and desiccation tolerant. However, further studies using more species are required to validate this approach. This may be an ecophysiological or genetic trait of these species. Therefore, this form of dry seed cryopreservation could form part of ex situ orchid seed conservation using a standard method. The methods developed here will store greater genetic diversity compared to what can be achieved with protocorms and are suitable for both asymbiotic and symbiotic recovery after cryopreservation. This will help reduce the time and cost of ex situ conservation, and help develop universal protocols for large genera, compared to custom protocols required for protocorm cryopreservation. [ABSTRACT FROM AUTHOR]