Semen cryopreservation and the conservation of endangered species

Currently, more than 16,000 plant and animal species are imminently threatened by extinction, often as a direct consequence of anthropogenic influences. One of the measures to halt that process is genetic resource banking. This short review focuses on mammal sperm cryopreservation in combination with assisted reproduction techniques. It summarizes general problems, recent developments, and currently applied protocols and gives an overview of hitherto existing successes of assisted reproduction measures in wild animals in the light of conservation efforts.     

濒危物种保护方法研究进展

对濒危物种的科学内涵、濒危机制和物种保护方法进行综述。具体阐述了种群生存力分析 (PVA)技术和复合种群理论(Meta- population)在濒危物种保护中的应用 ;总结了分子生物学方法在濒危物种保护中的作用 ;探讨了“3S”技术在濒危物种保护中的应用前景。分析和评价了各种保护方法的实用性和局限性 ,提出了各种保护方法在濒危物种保护中的应用前景和发展趋势。总之 ,物种保护方法的不断发展开创了保护生物学研究的新篇章 ,怎样运用交叉学科理论和综合应用各种保护方法探讨物种濒危机制、制定濒危物种有效管理和保护措施 ,是需要进一步解决的科学问题。     

Delimiting species boundaries and the conservation genetics of the endangered maritime ringlet butterfly (Coenonympha nipisiquit McDunnough)

Species delimitation is a difficult problem that has implications across organismal biology, yet no single method has proved wholly satisfactory. We tested the utility of combining species-delimitation methods based on phylogeny and gene flow statistics using two parapatric members of the Coenonympha tullia group as an example: the endangered maritime ringlet butterfly (Coenonympha nipisiquit McDunnough) and the common inornate ringlet butterfly (Coenonympha inornata Edwards). We reconstructed the phylogeny of the nearctic C. tullia-group taxa from mitochondrial DNA (mtDNA) sequences (cytochrome oxidase I and mitochondrial control region) to explore the ancestry of the C. nipisiquit lineage within the group. We investigated the extent of gene flow between the two taxa with F-statistics using 587 nuclear amplified fragment length polymorphism markers, accounting for the effect of potential scoring 'collisions' where a marker may represent more than one DNA fragment. Combining species-delimitation methods was especially effective because it uncovered both historical and recent evolutionary patterns. Phylogenetic analysis of mtDNA revealed the early divergence of C. nipisiquit from other C. tullia-group taxa, including the morphologically similar C. inornata. F-statistics and gene-by-gene introgression profiles demonstrated clear isolation between the two taxa and revealed strong population structure within C. nipisiquit. C. nipisiquit is the first taxon in the nearctic C. tullia group showing strong evidence of genetic isolation. The methods we used are relatively inexpensive and can be widely used to delimit taxonomic boundaries near the species level, both generally and in particular for taxa that may be targets of conservation efforts.     

Captive breeding of endangered fish Chitala chitala (Hamilton-Buchanan) for species conservation and sustainable utilization

Over the last few decades wild population of Chitala chitala (HamiltonBuchanan) has been declined more than 50% due to various reasons and is presently listed under endangered (EN) category due to reduced abundance. In the present communication wild C. chitala were collected from natural habitats and induced to spawn under captivity during July 2002 by injecting three different doses of synthetic hormone Ovaprim. Intramuscular injections were administered to fishes using three different doses (1.5, 1.0 and 0.5 ml/kg body weight). Artificial breeding pool was prepared for each set by encircling area (20 × 5 m) with mosquito net, where wooden country boat (8 × 4 × 2.5 feet with surface area 48.5 sq. feet) was placed inside the breeding pool. Distinct spawning behavior was noticed in the experimental sets with different hormonal dose whereas no spawning activity was noticed in control set. The fertilization rate varied from 48.8680.2% and total numbers of spawned eggs in two sets of experiments were estimated to be 81,034. The average number of eggs deposited 15 ± 2.1/square inches. The fertilized eggs were large in size (4.5 ± 0.05 mm), adhesive and attached to the hard substratum. The eggs hatch out between 168192 h after fertilization and about 33,639 hatchlings were produced. Newly hatched larvae measured 10.23 ± 0.03 mm and 0.031± 0.008 gm in weight and the mean diameter of yolk sac was 4.1 ± 0.08 mm. The yolk sac remains attached up to a week. The percentage survival of hatchlings varied from 42.2 to 65.60. Statistical analysis was worked out to determine the relation between the hormone dosage with different breeding parameters like latency period, fertilization rate, egg output, hatching rate and hatchling production.     

稀有濒危植物坡垒的迁地保护

国家二级保护树种坡垒（ＨｏｐｅａｈａｉｎａｎｅｎｓｉｓＭｅｒｒ．ｅｔＣｈｕｎ）原产海南岛。在西双版纳勐仑热带植物园迁地栽培,年平均增高０．４４～０．９ｍ,胸径加粗０．５４～１．３１ｃｍ,比原产地野生或其他迁地保护地区的植株生长快。花期８～１０月,果熟期翌年３～５月。种子发芽率达９６．７％。经多年观察,未见寒害和严重的病虫害。已繁衍了３代并扩大种植２８４株,说明坡垒在西双版纳热带植物园迁地保护获得成功。     

DNA分子标记技术在濒危物种保护中的应用

近20年来,随着分子生物学技术的迅猛发展,涌现出一批高效、可靠的DNA分子标记技术.本文论述了限制性片段长度多态性、微卫星DNA、随机扩增多态性DNA、扩增片段长度多态性等DNA分子标记技术的基本原理及技术特点;同时,介绍了DNA分子标记在濒危物种种群遗传学研究、致危因素分析及保护策略的制定等保护生物学方面的应用.     

Review of the biology,uses and conservation of the critically endangered endemic Himalayan species Nardostachys jatamansi (Caprifoliaceae)

Biodiversity and Conservation - The commercial demand for Nardostachys jatamansi in the global market has raised concern about its long-term sustainability. The genus Nardostachys is represented by...     

黄河流域濒危物种保护热点区与保护空缺识别

黄河流域具有重要的生物多样性保护意义,通过研究珍稀濒危物种分布热点区可为生物多样性保护提供依据。选取70种濒危维管植物和陆生脊椎动物,综合多来源的分布数据,运用物种分布模型Maxent模拟物种分布区,结合自然地理区划,计算保护价值,进行热点区分析,并结合国家级自然保护区和国家公园体制试点区的分布情况进行空缺分析。研究结果显示,黄河流域濒危物种分布主要呈现出南高北低、集中于山地的特征,热点区包括秦岭区域、太行山区域、子午岭-六盘山区域、陇中高原至松潘高原、祁连山、贺兰山和沿黄湿地等。在区分自然地理区后,现有的国家级自然保护区和国家公园体制试点区覆盖了热点区面积的13.89%,保护空缺主要出现于子午岭南部、六盘山南部、松潘高原南部和拉脊山等。建议在推进黄河流域生态保护和高质量发展中将濒危物种热点区考虑在内,对黄河流域自然保护地体系进行优化,并针对黄河流域的三个自然地理分区提出了相应的保护建议。此外,研究发现,在进行热点区分析时,考虑自然地理区域划分,并综合多类群叠加和单一生物类群的分析结果进行统筹考虑,可能会更好满足生物多样性就地保护需求。     

Reintroduction of the endangered and endemic plant species Cochlearia bavarica—Implications from conservation genetics

Population reintroduction is a common practice in conservation, but often fails, also due to the effects of inbreeding or outbreeding depression. Cochlearia bavarica is a strongly endangered plant species endemic to Bavaria in Germany, constantly declining since the late 1980s. Therefore, population reintroduction is intended. In this study, we analyzed genetic diversity within and genetic differentiation between all 32 remnant populations of the species in Swabia and Upper Bavaria using amplified fragment length polymorphisms. Our aim was to increase reintroduction success by providing data to avoid negative effects of inbreeding and outbreeding and to preserve the natural genetic pattern of the species. Genetic diversity within populations was low but similar to other rare and endemic species and varied strongly between populations but did not depend on population size. Our analysis revealed a strong geographic pattern of genetic variation. Genetic differentiation was strongest between Swabia and Upper Bavaria and at the population level, whereas differentiation between subpopulations was comparatively low. Isolation by distance and genetic differentiation was stronger among populations from Upper Bavaria than from Swabia. From the results of our study, we derived recommendations for a successful reintroduction of the species. We suggest using rather genetically variable than large populations as reintroduction sources. Moreover, the exchange of plant material between Swabia and Upper Bavaria should be completely avoided. Within these regions, plant material from genetically similar populations should preferably be used for reintroduction, whereas the exchange among subpopulations seems to be possible without a negative impact on genetic variation due to natural gene flow.     

濒危植物紫纹兜兰群落特征及其保护

对广东省河源市紫金县白溪自然保护区紫纹兜兰（Paphiopedilum purpuratum（Lindl.）Stein）所在的2个群落进行了样方调查,分析了其生境、物种组成、群落区系特点及群落结构特征。结果显示,调查样地内共有维管束植物47科79属94种。属的区系成分中泛热带和热带亚洲成分占主要地位,占非世界分布总属数的84.3%,具有热带向亚热带过渡的性质。群落1中扇叶铁线蕨（Adiantum flabellulatum Linn.）和中华苔草（Carex chinensis Retz.）为优势种,紫纹兜兰仅为伴生种,群落2中紫纹兜兰为优势种。两群落均以小高位芽植物为主,但缺乏大高位芽植物、地面芽植物及一年生草本,而藤本植物的丰富则进一步加大了林下的郁闭度。综合生境因素比较两个群落表明,紫纹兜兰对环境具有极强的依赖性,对坡度坡向及环境湿度有一定的要求,对伴生植物并无明显的选择性。建议加强对紫纹兜兰的就地保护,同时加大对其栽培繁殖技术的研究。     

Genetic diversity and structure in fragmented populations of the endangered species Ranunculus cabrerensis (Ranunculaceae): implications for conservation

Ranunculus cabrerensis is an endemic and endangered species of the Northwestern Iberian Peninsula. The molecular markers AFLP and ISSR were used to investigate the genetic diversity and population structure of four populations across its known distribution. Fifteen selective primer combinations of AFLP and seventeen ISSR primer combinations produced a total of 2830 and 103 unambiguously repeatable fragments respectively, of which 97.57 and 81.38% were polymorphic for both markers. The genetic diversity of R. cabrerensis at species level was high (H _E = 0.294 by ISSR and H _E = 0.191 by AFLP) and differentiation between sampled locations was also relatively high (G _ST = 0.316 and 0.158 by ISSR and AFLP analysis respectively) compared to other studies of endangered and rare species using the same techniques. The analysis of molecular variance (AMOVA) indicated that the main genetic variation was within sampled locations (73% by AFLP; 52% by ISSR), even though the variation among locations was also significant. Principal Coordinates, NeighborNet and Bayesian analyses revealed a weak but significant relationship between the genetic structures of different populations in R. cabrerensis, with gene flow acting as a homogenizing force that prevents stronger differentiation of populations. Finally, suggestions for conservation strategies to preserve the genetic resources of this species are outlined.     

The importance of genetic cluster recognition for the conservation of migratory fish species: the example of the endangered European huchen Hucho hucho (L.)

European huchen Hucho hucho (L.) is an endangered flagship species, which is endemic to the Danube drainage in central Europe. To date, no genetic information has been available as a basis for ongoing conservation and breeding programmes for the species. It is suspected that most populations in the wild share one common gene pool and that they exclusively depend on stocking with hatchery fish. In this study, highly variable microsatellite markers were established and the genetic diversity and differentiation from four important hatchery-reared stocks were compared with that of eight H. hucho populations sampled in the wild. Overall, eight genetic clusters with a moderate to very great degree of genetic differentiation and high assignment rates were identified. Each cluster contained individuals from two to 10 different populations and 9–100% of specimens from hatchery stocks. It is proposed that genetic cluster-based management in the conservation of European huchen is advantageous compared with the consideration of single local populations. A combined approach of maintaining the evolutionary potential of wild populations and genetically variable hatchery stocks can maximize the conservation of the species' evolutionary potential.     

Evolutionary distinctiveness and conservation priority of the endangered Najas ancistrocarpa (Hydrocharitaceae)

Phylogeny is vitally important to evaluate evolutionary distinctiveness and conservation priorities of species. Najas ancistrocarpa is one of the rarest and least known East Asian species of the c. 30–40 species of the aquatic plant genus Najas (Hydrocharitaceae). We used a recently discovered collection of this nationally and regionally endangered species in Japan and East Asia to assess whether N. ancistrocarpa is a distinct species or an untypical form of other Najas species and to evaluate its conservation priority in a phylogenetic context. The sample set of our previous study, exploring worldwide phylogeny of the genus was used with N. ancistrocarpa to delimit species boundaries in Najas and obtain conservation priority scores, calculated by multiplying evolutionary distinctiveness and global extinction risks converted from Red List categories. We performed molecular phylogenetic analyses of plastid (ptDNA) and nuclear (ITS) DNA datasets, STACEY species delimitation analysis using the multilocus dataset and a measurement of conservation priority with Tuatara. Najas ancistrocarpa was consistently placed in ptDNA and ITS trees. Species delimitation analysis objectively assessed that N. ancistrocarpa is phylogenetically distinct. Phylogenetic conservation prioritization ranked N. ancistrocarpa the second highest priority species of the genus Najas. Overall, in terms of biodiversity conservation, N. ancistrocarpa represents an important Najas species in Japan and probably the world.     

A role for molecular genetics in the recognition and conservation of endangered species

Taxonomies based on morphological traits alone sometimes provide inadequate or misleading guides to phylogenetic distinctions at the subspecies and species levels. Yet taxonomic assignments inevitably shape perceptions of biotic diversity, including recognition of endangered species. Case histories are discussed in which the data of molecular genetics revealed prior systematic errors of the two possible kinds: taxonomic recognition of groups showing little evolutionary differentiation, and lack of taxonomic recognition of phylogenetically distinct forms. In such cases, conservation efforts for 'endangered species' can be misdirected with respect to the goal of protecting biological diversity.     

Micropropagation of the endangered species Malus niedzwetzkyana for conservation biodiversity in Kazakhstan

In Vitro Cellular & Developmental Biology - Plant - Biodiversity conservation requires advanced and effective ex situ plant propagation techniques. The present study was conducted to optimize...     

内蒙古珍稀濒危植物濒危现状及优先保护评估

以《中国珍稀濒危保护植物名录》、《中国植物红皮书》、《国家重点保护野生植物名录(第一批)》、《中国物种红色名录》、《中国生物多样性红色名录(高等植物卷)》、《内蒙古珍稀濒危保护植物名录》和《内蒙古珍稀濒危植物图谱》中127种内蒙古珍稀濒危植物为研究对象,通过资料收集及专家咨询,构建了内蒙古珍稀濒危植物受威胁等级、优先保护评估体系.建立了濒危系数、遗传价值系数、利用价值系数、生境系数、繁殖系数5项准则,准则下共设17个指标;运用层次分析法确定指标体系权重,计算出珍稀濒危植物濒危等级及优先保护级别.结果表明: 极危种2种、濒危种13种、易危种37种、近危种44种、无危种31种,分别占总数的1.6%、10.2%、29.1%、34.7%、24.4%.其中,受威胁种(极危、濒危和易危种)共52种,占总种数的40.9%.一级保护植物35种、二级保护植物72种、三级保护植物20种,分别占总数的27.6%、56.7%、15.8%.本评估结果与《中国生物多样性红色名录(高等植物卷)》、《内蒙古珍稀濒危保护植物名录》相比,有75种植物的濒危等级和62种植物的保护级别发生了变化.其中新增了9种植物的濒危等级评估和32种植物的保护级别.     

A resource-based conservation approach for an endangered ecotone species: the Ilex Hairstreak (Satyrium ilicis) in Flanders (north Belgium)

Ecotones are often species-rich and harbour specific resources and environmental conditions for invertebrates. Despite their functional significance for conservation, they are often not explicitly included in biotope typologies relevant to conservation policy and management (e.g., the European Habitats Directive). The Ilex Hairstreak (Satyrium ilicis) is a species of European conservation concern and a typical ecotone species inhabiting gradients from open (e.g., heathland, grassland) to closed vegetation (e.g., woodland). Here, we investigated its occurrence and habitat use at different spatial scales in Flanders (north Belgium). At a regional scale, species distribution modelling predicted 1,152 grid cells of 1 × 1 km² to be suitable of which 190 were presumed to occur within colonization capacity (±2.5 km). At a local scale, adult butterflies were more abundant on sites sheltered by bushes and small trees and with nectar sources in the vicinity of tall oak trees (mate locating sites). For egg-laying, females preferred oaks of intermediate height (50–150 cm) with many low branches at some distance from the nearest woodland edge (12 m). Additionally, Alder Buckthorn (Frangula alnus) was abundant as well as a herb layer of 10–15 cm. 73 % of the eggs were parasitized and parasitism occurred more often within sites where small oaks were very abundant. Making use of our results, we suggest conservation measures at different scales for this endangered ecotone species: policy measures at a regional level to delineate functional conservation units and local management measures using a resource-based approach.