<Emphasis Type="Italic">In vitro</Emphasis> and cryogenic preservation of plant biodiversity in Brazil

The Brazilian biomes (Amazon, Atlantic Forest, Cerrado, Caatinga, Pantanal, and Pampa) comprise one of the highest levels of plant diversity in the world; however, non-sustainable practices, deforestation, and land use have resulted in significant losses and fragmentation of the native forests. These ecosystems are now threatened and protection of their native plants through ex situ conservation is an urgent necessity. Cryopreservation and in vitro conservation are complementary options for securing and protecting Brazil’s native plant species because their potential economic value is critically important to develop strategies that will (1) support their sustainable utilization, (2) protect against the over-exploitation of species growing in natural habitats, and (3) conserve the genetic diversity of germplasm from species of different provenances. Biotechnological approaches will help to address future economic and environmental demands placed upon already at-risk species. Conserving seed germplasm ex situ provides an additional safeguard against the risks (e.g., loss due to disease, climate change) of field conservation. Moreover, seed banks and cryobanks permit the long-term conservation of a wider genetic base; this offsets the labor and space intensive costs of conserving in the active growing state. This paper is a compilation of the current status of strategies applied for conserving Brazilian native plant species.     

The use of biotechnology in the conservation of tropical germplasm

The rapid erosion of the genetic diversity of both wild and cultivated plants has recently attracted more and more international concern. As a consequence, germplasm conservation techniques have become accessible to protect the third world's germplasm diversity. Several approaches are used for germplasm conservation and utilization. The evaluation and use of these techniques has to be focussed with one important objective; to preserve with as much integrity as possible, the genetic variability of the selected species.Questions have risen about the best way to preserve genetic diversity of the humid tropics, since it is being recognized that these conditions hold much of the world's gene pool of plants. Seed banks are considered the best system for orthodox seed storage. For many tropical species that cannot support dehydration, tissue culture and cryoconservation are the best alternative. Important considerations regarding the adaptation of modern techniques to tropical species have to be carefully analyzed. Quantitative studies of genetic indices of somaclonal variation, or types or mutant plants can be observed.Consideration of plant germplasm as a base for genetic improvement has come a long way, but much remains to be done, particularly, with the plant diversity that exists in the tropics. All efforts to conserve and use genetic resources will contribute to the benefit of future human generations.     

乡土植物生活型构成对川渝地区边坡植被水土保持效益的影响

乡土植物灌木化建植是高速公路边坡防护的重要生态模式。为了探讨生态恢复过程中乡土植物的生活型构成对边坡植被水土保持效益的影响,揭示乡土植物生活型-物种多样性-生态系统功能间的偶合关系,借助3个物种配置试验,于2009年4月构建了以草本、灌木或乔木为主体的草本型、灌木型、乔木型绿化配置及草-灌-乔混合型试验区。自建植次年(2010年)起,对试验区进行持续5a生态监测。结果表明:1)边坡植被的物种丰富度与乡土植物的生活型有关,呈现乔木型灌木型草-灌-乔混合型草本型趋势;2)植被的物种多样性(Shannon-Wiener指数、Pielou指数)与乡土植物生活型构成及建植年限有关:建植后第1、第2年,多样性水平呈草本型草-灌-乔混合型灌木型乔木型变化趋势;自建植后第三起(2012—2014年),呈草-灌-乔混合型草本型灌木型乔木型波动;3)植被的水土保持性能(径流系数、侵蚀模数)与群落的物种多样性密切相关:多样性水平越高,水土保持性能越强。可见,乡土植物的生活型构成对提高边坡植被的物种多样性、改善生态性能至关重要。     

Ex Situ Conservation Priorities for the Wild Relatives of Potato (Solanum L. Section Petota)

Crop wild relatives have a long history of use in potato breeding, particularly for pest and disease resistance, and are expected to be increasingly used in the search for tolerance to biotic and abiotic stresses. Their current and future use in crop improvement depends on their availability in ex situ germplasm collections. As these plants are impacted in the wild by habitat destruction and climate change, actions to ensure their conservation ex situ become ever more urgent. We analyzed the state of ex situ conservation of 73 of the closest wild relatives of potato (Solanum section Petota) with the aim of establishing priorities for further collecting to fill important gaps in germplasm collections. A total of 32 species (43.8%), were assigned high priority for further collecting due to severe gaps in their ex situ collections. Such gaps are most pronounced in the geographic center of diversity of the wild relatives in Peru. A total of 20 and 18 species were assessed as medium and low priority for further collecting, respectively, with only three species determined to be sufficiently represented currently. Priorities for further collecting include: (i) species completely lacking representation in germplasm collections; (ii) other high priority taxa, with geographic emphasis on the center of species diversity; (iii) medium priority species. Such collecting efforts combined with further emphasis on improving ex situ conservation technologies and methods, performing genotypic and phenotypic characterization of wild relative diversity, monitoring wild populations in situ, and making conserved wild relatives and their associated data accessible to the global research community, represent key steps in ensuring the long-term availability of the wild genetic resources of this important crop.     

Importance of <Emphasis Type="Italic">in vitro</Emphasis> technology to future conservation programmes worldwide

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.     

Impact of land use on plant biodiversity and measures for biodiversity conservation in the Loess Plateau in China – a case study in a hilly-gully region of the Northern Loess Plateau

The Loess Plateau is a special natural–cultural unit in northern China. Intensive land use in the past has had, and forestation and grass planting at present will have inevitable impacts on plant biodiversity in the Loess Plateau. Based on the analysis of floristic features within three sampling sites with different land use practices and analysis of species richness among different land use types, we discuss impacts of land use on species richness and floristic features in the Northern Loess Plateau. The results drawn from this case study are as follows: (1) It appears that forestation and grass planting have had a positive influence on the local species diversity, but they have contributed little to the native vegetation in terms of conserving its floristic features. (2) Caragana intermedia shrubland, Pinus tabulaeformis forestland, and natural grassland have made important contributions to supporting indigenous species and maintaining local plant biodiversity. (3) There is a significant positive correlation between land use diversity and species richness. These results imply that practicing biodiversity conservation in situ is feasible and the suitable choice for the Loess Plateau. Concrete measures for biodiversity conservation in the area can include setting up small nature reserves and diversifying land use patterns to maintain as much habitat as possible for native vegetation. The artificial Hippophae rhamnoides shrubland should not be further promoted, considering its negative influence on biodiversity conservation.     

Endemic wild potato (Solanum spp.) biodiversity status in Bolivia: Reasons for conservation concerns

Crop wild relatives possess important traits, therefore ex situ and in situ conservation efforts are essential to maintain sufficient options for crop improvement. Bolivia is a centre of wild relative diversity for several crops, among them potato, which is an important staple worldwide and the principal food crop in this country. Despite their relevance for plant breeding, limited knowledge exists about their in situ conservation status. We used Geographic Information Systems (GIS) and distribution modelling with the software Maxent to better understand geographic patterns of endemic wild potato diversity in Bolivia. In combination with threat layers, we assessed the conservation status of all endemic species, 21 in total. We prioritised areas for in situ conservation by using complementary reserve selection and excluded 25% of the most-threatened collection sites because costs to implement conservation measures at those locations may be too high compared to other areas. Some 70% (15 of 21 species) has a preliminary vulnerable status or worse according to IUCN red list distribution criteria. Our results show that four of these species would require special conservation attention because they were only observed in <15 locations and are highly threatened by human accessibility, fires and livestock pressure. Although highest species richness occurs in south-central Bolivia, in the departments Santa Cruz and Chuquisaca, the first priority area for in situ conservation according to our reserve selection exercise is central Bolivia, Cochabamba; this area is less threatened than the potato wild relatives’ hotspot in south-central Bolivia. Only seven of the 21 species were observed in protected areas. To improve coverage of potato wild relatives’ distribution by protected areas, we recommend starting inventories in parks and reserves with high modelled diversity. Finally, to improve ex situ conservation, we targeted areas for germplasm collection of species with <5 accessions conserved in genebanks.     

Is It Still Necessary to Continue to Collect Crop Genetic Resources in the Mediterranean Area? A Case Study in Catalonia

Crop genetic resources have been extensively collected in Europe in the last century, creating large, publicly available ex situ collections. While this huge genetic diversity is often underutilized, in recent decades, several initiatives have emerged at the local level to collect germplasm cultivated on farm. Uncoordinated actors often carry out these collecting missions without considering previously collected data. To explore whether new collecting missions are likely to be worthwhile, we studied the crop genetic resources conservation network in Catalonia by analyzing the passport data and geographical distribution of germplasm stored in seed banks. Moreover, to determine whether this germplasm was representative of the diversity cultivated on farm, we performed new collecting missions in four randomly selected areas in the European Union’s Natura 2000 network and compared the results with the ex situ databases. Seed banks currently hold a large germplasm collection (2931 accessions), although most materials are conserved in private collections without regulated systems for seed regeneration and are not present as duplicates in the National Inventory. One important shortcoming of the ex situ network is that the germplasm conserved ex situ shows a low geographical coverage, representing only 35.3% of the municipalities in Catalonia. Our new missions allowed us to collect 234 accessions, mostly tomatoes (17.5%) and beans (16.2%). The ecological indicators’ richness (both at species (S) and variety (V) levels), total abundance (A), and the Shannon-Weaver diversity index calculated at species (H2, considering the different accessions of each variety as a single population) and variety levels (H3, considering the intra-varietal genetic diversity) were higher in the newly collected germplasm than in the ex situ collections, suggesting that seed banks do not accurately represent the genetic diversity still cultivated on farm. Moreover, some important landraces from each area were absent or underrepresented in the ex situ collections. Thus, it is necessary to continue to devote efforts to collecting germplasm; better organization between actors and targeting specific species/varieties can increase the efficiency of new collecting missions. As a conclusion, we propose different criteria to guide new missions and to improve the network’s conservation activities.     

Can Biospecimen Science Expedite the Ex Situ Conservation of Plants in Megadiverse Countries? A Focus on the Flora of Brazil

Increasing the number of species conserved ex situ in Megadiverse countries is a major task exacerbated by many intricate factors including: biome complexity, wide range of biodiversity and an incomplete knowledge of life cycles, reproductive strategies, adaptations and species interactions. Although, establishing safe reserves is a crucial conservation measure their security and effective maintenance can be unfavourably compromised by climate change and the risks incurred by socioeconomic instability and changes in land use. Anthropogenic impacts, non-sustainable practices and habitat erosion have motivated current international efforts which focused on Brazil as host of ‘Rio+20’ the United Nation's twentieth anniversary conference on sustainable development. The revised targets of the Global Strategy for Plant Conservation (GSPC) are responses to species decline and realizing Target 8, which concerns ex situ conservation, places the heaviest burdens on countries that are custodians of the highest levels of global biodiversity. At the scientific level, ex situ conservation of endemic species in genebanks is often hindered by a lack of information about molecular genetics and problematic (recalcitrant) storage behaviors that restrict the preservation of flora native to Megadiverse countries. The potential for applying the ‘Biospecimen Science’ paradigm in expediting conservation in biodiversity-rich biomes is considered using Brazil as an exemplar of a Megadiverse country. The impacts of process chains on the quality of preserved plant germplasm and using evidence-based research to improve conservation outcomes, risk and quality management systems are appraised. The Biospecimen Science approach is not intended to displace conventional conservation practices but rather, to enhance their effectiveness in terms of the scale and efficiency of their scientific and technical operations.     

Ex situ conservation of plant germplasm using biotechnology

Conservation of plant genetic resources attracts more and more public interest as the only way to guarantee adequate food supplies for future human generations. However, the conservation and subsequent use of such resources are complicated by cultural, economical, technical and political issues. Over the last 30 years, there have been significant increases in the number of plant collections and in accessions in ex situ storage centres throughout the World. The present review is of these ex situ collections and the contribution biotechnology has made and can make to conservation of plant germplasm. The applications and limitations of the new, molecular approaches to germplasm characterization are discussed. In vitro slow growth is used routinely for conserving germplasm of plants such as banana, plantain, cassava and potato. More recently, cryopreservation procedures have become more accessible for long-term storage. New cryopreservation techniques, such as encapsulation-dehydration, vitrification and desiccation, lengthen the list of plant species that can not only tolerate low temperatures but also give normal growth on recovery. Extensive research is still needed if these techniques are to be fully exploited.V.M. Villalobos is with the Food and Agriculture Organization of the United Nations, Viale delle Terme di Caracalia, 00100 Rome, Italy. F. Engelmann is with the International Plant Genetic Resources Institute (IPGRI), Via delle Sette Chiese 142, 00145 Rome, Italy.     

Strategies for agrobiodiversity conservation and promotion: a case from Yunnan, China

This paper deals with strategies for agrobiodiversity conservation and promotion based on studies on four plant groups (selected from 50 plant groups) occurring in the Yunnan Province of China. These plants are edible konjac (Amorphophallus), medicinal Paris, Musella lasiocarpa and wild tea (Camellia sinensis and its wild relatives), including their cultivars and wild populations. After analyzing the target plants, we conclude that the following strategies should be adopted to conserve and promote agrobiodiversity: (1) in situ conservation of agrobiodiversity, including habitat protection of wild populations, maintenance of native species and varieties in traditional agroecosystems, and relevant environmental education; (2) ex situ conservation and promotion of agrobiodiversity, including establishment of living collections and germplasm banks, and introduction of species and varieties into agroecosystems for agricultural practice; and (3) promotion and conservation of agrobiodiversity through sustainable uses, including technique development of propagation, cultivation, pest and disease control, on farm and off farm management, and other activities such as new variety breeding and scientific studies. Strategies developed here will be helpful to conserve and promote agrobiodiversity at agroecosystem, species, variety or landrace, and management system levels.     

Cultural responsibility in the preservation of local economic plant resources

Understanding the relationships between indigenous people and their threatened economic plants can aid the conservation effort on many levels. Understanding ethnic perceptions of the taxon is critical toin situ andex situ conservation projects and enhances the accompanying educational effort. Examples are discussed from the experience of a grassroots conservation group in southwestern United States, Native Seed/SEARCH. Four levels of economic plant vulnerability are examined among 1) wild-harvested plants, 2) husbanded wild plants, 3) domesticates, and 4) wild relatives of domesticates. Legal interpretations of endangered husbanded and domestic plants are discussed, and further documentation encouraged. Genetic dynamism of threatened indigenous crops is examined and the concept of Systems Conservation (i.e. the plant/human interactive systems) is introduced. Guidelines are offered for incorporating better cultural responsibility intoex situ conservation strategies. The concept of Biocultural Restoration is introduced with an example from an O'odham community. Examples are given of ways indigenous peoples and their knowledge can assist in the conservation effort.     

Gap analyses to support ex situ conservation of genetic diversity in Magnolia, a flagship group

Worldwide about one third of all plant species is estimated to be threatened with extinction. Plants are generally under-represented in conservation. However, the global strategy for plant conservation (GSPC) sets forth 16 targets to halt the current and continuing loss of plant diversity within a framework for actions at global, regional, national and local levels. Target 8 of the GSPC directs that at least 75 % of threatened plant species be present in ex situ collections by 2020, but this target is far from being met. By analyzing where gaps in ex situ collections and research exist relative to diversity hotspots of threatened species, it is possible to identify and prioritize action. We undertake a gap analysis of ex situ collection needs in the genus Magnolia, a relatively well-known and valuable genus. Results indicate three main areas of biodiversity (China, Vietnam and Colombia) for the endangered Magnolia taxa. The present paper provides a critical overview of recent conservation activities carried out in the genus Magnolia throughout the world, focusing on genetic diversity analyses of the species.     

Farmers Drive Genetic Diversity of Thai Purple Rice (Oryza sativa L.) Landraces

Purple or black rice (Oryza sativa L.) is a culturally important germplasm in Asia with a long history of cultivation in northern Thailand. Purple rice is identified by the color of the rice pericarp, which varies from purple to black with the accumulation of phenolic acids, flavonoids, and anthocyanins. In the present study, we assessed molecular variation within and between wetland purple rice landraces germplasm from northern and northeastern Thailand using 12 microsatellite loci. All purple rice varieties surveyed showed high levels of homozygosity within varieties and strong genetic differentiation among varieties, indicating the fixation of genetic differences among them. This pattern is consistent with purple rice farming practices in northern Thailand, where a small portion of harvested seed is selected and replanted based on farmers’ preferences. The reduced genetic diversity and high homozygosity observed for purple rice is also consistent with patterns expected for this inbreeding crop. Genetic differentiation among the varieties showed some degree of structuring based on their geographical origin. Taken together, these data highlight that the genetic diversity and structure of wetland purple rice landraces is shaped by farmer utilization and cultivation through local cultural practices, and that conservation should focus on ex situ conservation across its cultivation range, along with on-farm, in situ conservation based on farmers’ seed-saving practices. In situ conservation may prove especially valuable for preserving the genetic identity of local varieties and promote adaptation to local environments.

     

Genetic diversity and similarity revealed via molecular analysis among and within an in situ population and ex situ accessions of chiltepín (Capsicum annuum var. glabriusculum)

Molecular analysis of genetic diversity amongand within phenotypically similar wild Capsicum annuum var. glabriusculum(chile) populations revealed geneticdifferences among accessions spread over abroad geographic range. These chiles areregionally known as chiltepíns and are a 50metric ton per year wild harvest for the spiceindustry, as well as a genetic resource forcrop improvement. Understanding geneticvariability in this species providesinformation related to conservation efforts. The objective of this research was to surveygenetic diversity among and within an insitu population and ex situ accessionsof chiltepíns. Random AmplifiedPolymorphic DNA (RAPD) molecular markers wereused to study the genetic structure of an in situ population found at the nothernmostrange of this species and ex situaccessions collected from Mexico and Guatemala. Novel genetic variation was found in both thein situ northern disjunct population, aswell as some ex situ accessions, thussupporting conservation of this species viaboth in situ and ex situ strategies The evidence presented here supports effortsto conserve outlier populations via insitu management practices.     

Conservation genetics of two island endemic Ribes spp. (Grossulariaceae) of Sardinia: survival or extinction?

Measuring levels of population genetic diversity is an important step for assessing the conservation status of rare or endangered plant species and implementing appropriate conservation strategies. Populations of Ribes multiflorum subsp. sandalioticum and R. sardoum, two endangered endemic species from Sardinia, representing the whole genus on the island, were investigated using ISSR and SSR markers to determine levels and structure of genetic variability in their natural populations. Results indicated medium to low genetic diversity at the population level: Nei's gene diversity for ISSR markers ranged from 0.0840 to 0.1316; the expected heterozygosity (H_E) for SSR ranged from 0.4281 to 0.7012. In addition, only one remnant population of R. sardoum showed a high level of inbreeding, in accordance with its very small size. Regarding the structure of the six R. sandalioticum populations, both principal coordinates analysis (PCoA) and STRUCTURE analysis of ISSR and SSR data highlighted low population structure, although two populations appeared to be clearly distinct from the others. The genetic pattern of the two taxa associated with their different ecological positions indicated resilience of R. sandalioticum populations in fresh and humid habitats and uncertain future resistance for the residual R. sardoum population in xeric calcareous stands. Hence, this study highlights the importance of an integrated conservation approach (genetic plus in situ and ex situ conservation studies/measures) for activating management programmes in these endemic and threatened taxa that can be considered as crop wild relatives of cultivated Ribes species.     

The contribution of the Millennium Seed Bank Project to <Emphasis Type="Italic">ex situ</Emphasis> plant conservation in Chile

For a long time in situ conservation has been the main approach used to protect Chilean plant diversity. However, due to the high level of endemism of its flora (50%) and an increasing human impact on wild areas, ex situ conservation has become an urgent requirement to avoid the extinction of plant populations and species. Since 2001, the Instituto de Investigaciones Agropecuarias (INIA), Chile, has been working in partnership with the Royal Botanic Gardens Kew, (Kew) through the Millennium Seed Bank Project (MSB) with the objective of conserving 20% of the Chilean flora as seeds in long-term storage. This seed conservation effort has focussed mainly on the endangered and endemic plants of the Chilean drylands. Towards the end of the first phase of the MSB some 1482 seed collections representing 850 species and subspecies have been collected and safely preserved in the INIA Seed Base Bank and duplicated at Kew. Almost 70% of the total species collected are endemic to Chile and several of them are endangered. Additionally, seed germination research has been conducted for nearly 400 species and seed collections have been used to propagate several threatened species. Germination protocols have been published and disseminated online. Over 4,500 herbarium vouchers have been collected, largely duplicated at Kew and at the national herbarium in Chile. As a result of the inputs of INIA and the MSB, collaboration has been extended to other national stakeholders, mainly for plant taxonomy and seed collecting. In this context two training courses have been run for 70 staff/students. This training has contributed to the raising of general awareness of the need for the long-term protection of Chilean plant diversity and to demonstrate the key role that ex situ seed conservation can play in meeting this need.     

Molecular diversity and relationships among <Emphasis Type="Italic">Cymbidium goeringii</Emphasis> cultivars based on inter-simple sequence repeat (ISSR) markers

Spring orchid (Cymbidium goeringii) is a popular flowering plant species. There have been few molecular studies of the genetic diversity and conservation genetics on this species. An assessment of the level of genetic diversity in cultivated spring orchid would facilitate development of the future germplasm conservation for cultivar improvement. In the present study, DNA markers of intersimple sequence repeats (ISSR) were identified and the ISSR fingerprinting technique was used to evaluate genetic diversity in C. goeringii cultivars. Twenty-five ISSR primers were selected to produce a total of 224 ISSR loci for evaluation of the genetic diversity. A wide genetic variation was found in the 50 tested cultivars with Nei’s gene diversity (H = 0.2241) and 93.75% of polymorphic loci. Fifty cultivars were unequivocally distinguished based on ISSR fingerprinting. Cultivar-specific ISSR markers were identified in seven of 50 tested cultivars. Unweighted pair-group mean analysis (UPGMA) and principal coordinates analysis (PCA) grouped them into two clusters: one composed the cultivars mainly from Japan, and the other contained three major subclusters mainly from China. Two Chinese subclusters were generally consistent with horticultural classification, and the third Chinese subcluster contained cultivars from various horticultural groups. Our results suggest that the ISSR technique provides a powerful tool for cultivar identification and establishment of genetic relationships of cultivars in C. goeringii.     

Variation of plant diversity components in different scales in relation to grazing and climatic conditions

Background: Understanding the role of livestock grazing on plant diversity can be improved by an accurate measurement of diversity at all hierarchical scales due to the changeability of diversity components in space.

Aims: We evaluated the effects of grazing on plant species diversity at different scales of all common and rare species in two regions that have different climatic conditions (arid vs. semi-arid).

Methods: In each region, we collected abundant data of plant species from a nested sampling design that consisted of local (80 plots) and regional (16 sites) scales. We partitioned total species diversity (γ) into within plots (α_l), among plots (β_l) and among sites (β₂) using the additive partitioning.

Results: Diversity among sites contributed the most to total diversity for all and rare plant species in both regions. In addition, α₁ and β₁ diversities in ungrazed areas were greater than those in grazed areas for all and common species in both climates.

Conclusion: Abandonment of grazing after 10 years resulted in significant regeneration of common species at the local scale, with no change in rare species. We conclude that low grazing intensity is likely to be an important tool for conservation of plant diversity in which all scales should be considered.