Mating system and genetic diversity of a rare desert legume Ammopiptanthus nanus （Leguminosae）

Ammopiptanthus nanus is an endangered evergreen shrub endemic to the deserts of central Asia and plays an important role in delaying further desertification. We examined allozyme variation and AFLP diversity in A. nanus populations and investigated the mating system of this species using progeny arrays assayed for polymorphic allozyme loci. Mating system analysis in the Keyi＇eryongke＇er population showed low levels of outcrossing, and strong inbreeding depression. Low levels of genetic variation were detected at both population （allozyme, Pp=14.0%, A=l.14, He=0.031; AFLP, Pp=14.5%, Shannon＇s information index I=0.063） and species （allozyme, Pp=21.1%, A=1.21, He=0.040; AFLP, Pp=20.9%, I=0.083） levels; while moderate genetic differentiation existed among populations, as indicated by allozymes （Gsa-=0.081） and AFLP （GST=0.151-0.193）. Founder effect, bottlenecks in evolutionary history, the mixed mating system and co-ancestry may have influenced the level of genetic diversity in A. nanus. Markers of both types provide new insights for conservation management, indicating that the Biao＇ertuokuoyi and Keyi＇eryongke＇er populations should be given priority for in situ conservation and regarded as seed sources for ex situ conservation.     

Genetic diversity and geographic differentiation in endangered Ammopiptanthus (Leguminosae) populations in desert regions of northwest China as revealed by ISSR analysis

• Background and Aims The desert legume genus Ammopiptanthuscomprises two currently endangered species, A. mongolicus andA. nanus. Genetic variability and genetic differentiation betweenthe two species and within each species were examined. • Methods Inter-simple sequence repeat (ISSR) marker datawere obtained and analysed with respect to genetic diversity,structure and gene flow. • Key Results Despite the morphological similarity betweenA. mongolicus and A. nanus, the two species are geneticallydistinct from each other, indicated by 63 % species-specificbands. Low genetic variability was detected for both populationlevel (Shannon indices of diversity H_pop = 0·106, percentageof polymorphic loci P = 18·55 % for A. mongolicus; H_pop= 0·070, P = 12·24 % for A. nanus) and specieslevel (H_sp = 0·1832, P = 39·39 % for A. mongolicus;H_sp = 0·1026, P = 25·89 % for A. nanus). Moderategenetic differentiation was found based on different measures(AMOVA _ST and Hickory ^B) in both A. mongolicus (0·3743–0·3744)and A. nanus (0·2162–0·2369). • Conclusions The significant genetic difference betweenthe two species might be due to a possible vicariant evolutionaryevent from a single common ancestor through the fragmentationof their common ancestor's range. Conservation strategies forthese two endangered species are proposed.     

新疆沙冬青AM真菌群落结构与遗传多样性分析

以新疆乌恰县4个样地新疆沙冬青Ammopiptanthus nanus根围3个土层土壤为研究对象,利用Nested-PCR-DGGE技术,结合DGGE图谱分析、DNA克隆测序及系统发育分析,研究新疆沙冬青AM真菌菌群结构与遗传多样性特征,为新疆沙冬青根围功能菌群研究奠定基础。结果表明,不同样地和土层,AM真菌具有不同DGGE指纹图谱特征,AM真菌丰度、优势度及其多样性指数均有差异,其中上阿图什30–40cm土层AM真菌丰度和多样性指数最高,分别为20和3.48。DGGE条带测序和系统发育分析显示,全部序列归为2属14种,球囊霉属Glomus是其优势属,其中Uncultured Glomus(AB698612)和Uncultured Glomus(AB698616)普遍存在于4样地3个土层,Glomus sp.(FN429104)和Uncultured Glomus(KC797120)是上阿图什30–40cm土层特有种,Glomus sp.(EU332717),Rhizophagus intraradices(FR750206)和Uncultured Glomus(AB698616)是新疆沙冬青根围优势种。G.indicum和Rh.intraradices在形态学鉴定中未出现,说明分子生物学技术对真菌多样性研究起着补充和完善的作用。     

濒危植物小沙冬青研究进展

小沙冬青(Ammopiptanthus nanus (M. Pop.) Cheng f.)是荒漠干旱极端环境下唯一的常绿阔叶灌木,也是抗逆性研究的优秀指示植物材料。本文在查阅文献的基础上,对小沙冬青的群落生态学、解剖学、生殖生态学及遗传多样性、逆境胁迫下小沙冬青的生理及分子生物学响应方面的研究进行了综述,旨在为该植物资源的开发利用、荒漠区物种多样性的维持和生态环境建设提供借鉴。     

Genetic diversity in Cytisus aeolicus Guss. (Leguminosae), a rare endemite of the Italian flora

ABSTRACT

Cytisus aeolicus Guss, is an endemic plant restricted to the isles of Vulcano, Stromboli and Alicudi in the Aeolian archipelago. All known populations were assayed for genetic variability using enzyme polymorphism. Allozyme variation at 16 loci coding for 10 enzyme systems was examined. The great majority of loci turned out to be monomorphic or fixed heterozygous. The observed genetic depauperation is indicative of historical factors, such as the bottleneck effect associated with migration, and the founder effect in population re-establishment. The low genetic diversity is largely partitioned within rather than among populations, indicating that extant populations have not been established long enough for divergence to have occurred. The genetic uniformity, combined with the scattered distribution of small populations, and the negative shift of population size point to a critically endangered species. Strategies for conservation are considered.     

Reproductive biology and genetic diversity of a cryptoviviparous mangrove aegiceras corniculatum (Myrsinaceae) using allozyme and intersimple sequence repeat (ISSR) analysis

Mangroves consist of a group of taxonomically diverse species representing about 20 families of angiosperms. However, little is known about their reproductive biology, genetic structure, and the ecological and genetic factors affecting this structure. Comparative studies of various mangrove species are needed to fill such gaps in our knowledge. The pollination biology, outcrossing rate, and genetic diversity of Aegiceras corniculatum were investigated in this study. Pollination experiments suggested that the species is predominantly pollinator-dependent in fruit setting. A quantitative analysis of the mating system was performed using progeny arrays assayed for intersimple sequence repeat (ISSR) markers. The multilocus outcrossing rate (tm) was estimated to be 0.653 in a wild population. Both allozyme and ISSR were used to investigate genetic variation within and among populations. The combined effects of founder events and enhanced local gene flow through seedling dispersal by ocean currents apparently played an important role in shaping the population genetic structure in this mangrove species. Both allozyme variation (P = 4.76%, A = 1.05, HE = 0.024) and ISSR diversity (P = 16.18%, A = 1.061, HE = 0.039) were very low at the species level, in comparison with other woody plants with mixed-mating or outcrossing systems. Gene differentiation among populations was also low: allozyme GST = 0.106 and ISSR GST = 0.178. The unusually high genetic identities (0.997 for allozyme and 0.992 for ISSR loci), however, suggest that these populations are probably all descended from a common ancestral population with low polymorphism.     

Extraction,Separation and Measuration of the Thermal Hysteresis Activity of Antifreeze Proteins from Ammopiptanthus nanus (Leguminosae)

Antifreeze proteins (AFPs) , because of their ability of adsorbing onto the surface of ice crystals and modifying their growth , have been found in many different organisms that are exposed to chilling and freezing temperatures . It is very important to increase the resisting and coercing ability at low temperature . DE-52 ion- exchange cellulose was used to extract and separate the antifreeze proteins from Ammopiptanthus nanus leaves. Differential scanning calorimetry (DSC) was used to measure the thermal hysteresis activity (THA) . The results reveal that when the concentration of the protein is 20mg/ml , the THA of the AFPs is 0 . 46℃ . Its molecular weight is about 119. 24 kDa by sodium dodecyl sulfate polyacrylam ide gel electrophoresis (SDS-PAGE) .     

濒危植物矮沙冬青的等位酶多样性及居群分化

采用超薄平板微型聚丙烯酰胺等电聚焦技术,对珍稀濒危植物矮沙冬青(Ammopiptanthus nanus)6个自然居群的等位酶遗传多样性进行了初步研究。8个酶系统、l9个酶位点的检测结果表明,矮沙冬青具有较低的遗传变异水平。物种水平上的遗传多样性参数为：P=21．05％,A=1．21,Ht=0．040;居群水平上的平均遗传多样性为：P=14．04％,A=1．14(0．04),Ht=0．036(0．018),比长寿命多年生木本的狭域分布种和特有种的多样性水平都低。F-统计量分析显示,矮沙冬青在居群水平(F15=0．129)和总居群水平(FIT=0．202)上都表现为杂合子缺乏;Fsr=0．085表明居群问遗传分化较小,这与居群间较高的遗传一致度(I=0．997)的结果一致。据研究结果推测矮沙冬青进化历史上经历过遗传瓶颈,是造成遗传多样性水平偏低的重要原因。基于研究结果,提出应当重点保护遗传变异相对丰富的何依儿佣克尔(KE)居群,并且可将其作为引种的种源地。     

新疆沙冬青抗冻蛋白的提取分离及其热滞活性测定

抗冻蛋白被发现于低温环境下生存的生物体中,它能够吸附在冰晶表面并改变冰晶的生长形态,对于增加植物的抗低温胁迫能力有重要作用。用纤维素DE-52离子柱层析提取分离出了冬季新疆沙冬青(Ammopiptanthus nanus)叶片中的抗冻蛋白(AFPs)。差示扫描量热法(DSC)测定结果表明,当蛋白浓度为20mg/ml时,抗冻蛋白的热滞活性(THA)为0·46℃。经SDS-PAGE电泳分析,此抗冻蛋白的分子量为119·24kDa。     

中华水韭遗传多样性的RAPD分析

采用RAPD方法对珍稀濒危植物中华水韭(Isoetes sinensis)4个自然居群的48个样品进行了DNA多态性分析。从60个随机引物中筛选出14个有效引物,共产生124条DNA片段,其中72条为多态性条带,总的多态位点百分率(PPB)为58．06％。各居群间多态位点百分率差异显著(0．81％-12．90％)。AMOVA分析结果表明,4个居群间基因分化系数Фst=0．5894,即遗传变异中有相当一部分来源于群体间(58．94％)。日益缩小的种群规模而导致的居群内近交和遗传漂变的发生以及居群间有限的基因交流可能是中华水韭目前遗传结构的主要成因。鉴于目前中华水韭居群内个体数偏少、遗传多样性较低的现状,建议对其进行就地保护并保护尽可能多的生境,对不同自然居群内的个体进行植株相互移栽和育苗移栽,以提高不同居群间的基因交流,尽可能地保护中华水韭的遗传多样性。     

Genetic structure of the rare and endangered Hieracium wiesbaurianum group (Asteraceae) in Bavaria

The rare and endangered Hieracium wiesbaurianum species group shows a scattered relictual distribution in Bavaria. Recently, a couple of populations were discovered which clearly differ from all other populations. If these must be considered as taxonomically independent units, they would be of crucial conservation interest, because of the sole responsibility that Bavaria has for these worldwide endemics. We therefore analysed the genetic structure of H. wiesbaurianum in a comparative approach. Our analysis comprised 37 populations of 13 taxa of H. wiesbaurianum, H. bifidum and H. laevigatum, including three potentially new taxa. We applied amplified fragment length polymorphism (AFLP) analysis and observed only limited genetic variation within populations and taxa. Nevertheless, each studied individual exhibited a unique genotype. An analysis of molecular variance revealed high levels of genetic variation between taxa, but populations were genetically less different. The clear genetic differentiation between the studied taxa was supported by neighbor‐joining cluster analyses and principal coordinate analyses in which every individual was clearly assigned to its respective taxon. The three potentially new taxa were genetically as well differentiated as the other taxa included in our study. This supports the assumption that they should be treated as taxonomically independent units of high conservation interest. Therefore, the genetic analysis confirmed the morphologically based classification of the studied Hieracium taxa. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 177 , 112–123.     

Conservation genetics of the rare and endangered Leucopogon obtectus (Ericaceae)

Leucopogon obtectus Benth. is a declared rare species found in the kwongan vegetation in Western Australia. Plants on a mineral sand mine and the rehabilitation area are subject to disturbance. Genetic diversity was examined within and among all known populations using random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphisms (AFLPs) for conservation. Both molecular markers revealed a high percentage (> 89%) of polymorphic markers and a high mean genetic distance among individuals (D = 0.3). Analysis of molecular variance showed that 86.7% (RAPD) and 89.7% (AFLP) of variability was partitioned among individuals within populations. Exact tests showed no significant population differentiation. The analyses indicated that L. obtectus exhibits high levels of genetic diversity despite small population sizes. The high levels of variability among individuals and the lack of clear population differentiation suggest that this species comprises a single, genetically diverse group. Conservation and management of L. obtectus should concentrate on maintaining the high levels of genetic variability through mixing genotypes and promoting outcrossing.     

Genetic diagnosis of a rare myrmecochorous species,Plagiorhegma dubium (Berberidaceae): Historical genetic bottlenecks and strong spatial structures among populations

Distribution of genetic variation over time and space is relevant to demographic histories and tightly linked to ecological disturbances as well as evolutionary potential of an organism. Therefore, understanding the pattern of genetic diversity is a primary step in conservation and management projects for rare and threatened plant species. We used eight microsatellite markers to examine the level of genetic diversity, spatial structure, and demographic history of Plagiorhegma dubium, a rare myrmecochorous herb, populations sampled across northeast Asia and Siberia. We found low within‐population genetic variation associated with historical bottlenecks. Although pairwise F_ST values were not much higher than the ones found in similar life form species, STRUCTURE and PCoA revealed a clear broadscale spatial pattern of genetic structure. Bayesian clustering (best K = 6) and PCoA identified three populations that are distinctive from neighboring populations in the Korean peninsula, which suggests potential units for conservation and management plans in Korea. MIGRATE‐N and BAYESASS showed that both contemporary (0.003–0.045) and historical migration rates (2 × e^?5?4.6 × e^?4) were low. Our findings provide a good example, where genetic considerations should be integrated for conservation and management plans of rare and threatened species.     

夏腊梅的遗传多样性及其保护

夏腊梅（Sinocalycanthus chinensis)是国家二级保护的珍稀濒危植物,夏腊梅属（Sinocalycanthus)的唯一代表,仅间断分布于我国浙江省临安市天台县极狭小的范围内,本文采用等位酶淀粉凝胶电泳技术对采自上述两地的天然居群和天目山自然保护区引种的人工居群的553个样品进行了遗传多样性检测,并与浙江腊梅（Chimonan-thus zhejiangensis)作对比,检测结果表明夏腊梅的遗传多样性极低,从14个酶系统检测到的23个位点看,在物种水平上每位点的等位基因平均数（A）为1.2,多态位点（P）占21.7%,观察杂合度（Ho)为0.010。在居群水平上,A=1.0-1.1,P=0-13.0%,Ho=0-0.014。而对照种浙江腊梅杭州植物园人工居群的上述指标分别为A=1.5,P=39.1%,Ho=0.071。夏腊梅的2个自然居群之间在Mdh-4,Pgd-3和Sod-1发生显著的分化,但居群内亚居群间几乎没有分化,在天目山自然保护区引种的人工居群中没有检测到多态性,说明作为迁地保护的天目山自然保护区人工居群并没能有效地保护夏腊梅的遗传多样性,由于目前自然保护区基本上采用不加人为干预的经营方式,划人龙塘山自然保护区内的自然亚居群会因为森林的自然演替而灭绝,所以,自然保护区目前的这种经营方式不适合对夏腊梅的保护,夏腊梅的例子说明,当我们对保护对象的生物学特性缺乏认识时,我们既不知道应该保护什么,也不知道应该如何保护。     

水杉孑遗居群AFLP遗传变异的空间分布

本研究采用空间自相关分析方法对水杉 (Metasequoiaglyptostroboides)孑遗居群AFLP遗传变异的空间结构进行了研究 ,以探讨水杉孑遗居群遗传变异的分布特征及其形成机制。根据 6对AFLP选择性引物扩增的 46个多态性位点 ,选择了其表型频率在 2 5 %～ 75 %的 2 7个AFLP标记 ,运用等样本频率方法和等地理距离间隔方法分别对 3 9株和 3 7株原生母树进行了空间自相关系数Moran’sI值计算。结果表明 :水杉孑遗居群缺乏空间结构 ,绝大多数AFLP位点变异为随机分布的空间模式 ,但也有少数位点存在显著性随机相关 ,在 4～ 8km地理距离间隔显示负相关 ,说明该间隔可能是水杉孑遗居群的部分基因交流的有效屏障。水杉原生母树分布存在 12～ 2 8km的明显距离间隔空挡 ,说明人类从迁入该区域起就影响着水杉孑遗居群的原始生境 ,导致其生境片断化、景观破碎 ,进而形成岛屿状分布格局 ,并引起了水杉残留居群的随机遗传漂变。根据本研究结果 ,结合水杉孑遗居群较低的遗传多样性 ,分析探讨了水杉孑遗居群濒危的机理 ,并提出了相应的保育策略 ,为水杉的有效保育提供了科学依据     

Genetic diversity of the endangered Chinese endemic herb Primulina tabacum (Gesneriaceae) revealed by amplified fragment length polymorphism (AFLP)

Primulina tabacum Hance, is a critically endangered perennial endemic to limestone area in South China. Genetic variability within and among four extant populations of this species was assessed using AFLP markers. We expected a low genetic diversity level of this narrowly distributed species, but our results revealed that a high level of genetic diversity remains, both at population level (55.5% of markers polymorphic, H _E = 0.220, I _S = 0.321), and at species level (P = 85.6% of markers polymorphic, H _E = 0.339, I _S = 0.495), probably resulting from its refugial history and/or breeding system. High levels of genetic differentiation among populations was apparent based on Nei’s genetic diversity analysis (G _st=0.350). The restricted gene flow between populations is a potential reason for the high genetic differentiation. The population genetic diversity of P. tabacum revealed here has clear implications for conservation and management. To maintain present levels of genetic diversity, in situ conservation of all populations is necessary.     

濒危植物元宝山冷杉的遗传多样性研究

元宝山冷杉(Abies yuanbaoshanensis)是仅分布于广西融水县元宝山的珍稀濒危物种。本研究采用AFLP分子标记分析了元宝山冷杉种群的遗传多样性。取元宝山冷杉种群43棵植株作为研究材料,4对引物组合用于AFLP扩增,共得到261条DNA扩增带。分析结果：元宝山冷杉种群的多态位点百分比率为50．96％,Nei's基因多样性为0．1510,Shannon多样性指数为0．1735。用NTSYS软件计算各个样品间的相似性系数,并用UPGMA法基于相似性系数进行了聚类分析。研究结果表明：元宝山冷杉种群的遗传多样性水平低,种群内个体间的相似性系数很大,无明显亚种群间的遗传分化。要保护或保存这个物种的遗传资源,建议应该选择种群内尽可能多的个体进行保护。     

Comparison of genetic diversity between narrowly endemic shrub Menziesia goyozanensis and its widespread congener M. pentandra (Ericaceae)

Menziesia goyozanensis, a Japanese kindof fool's huckleberry, is a shrub speciesendemic only to Mt. Goyo in northern Japan.This species is listed as `criticallyendangered' in the Red Data Book of wild plantsin Japan, and only a single population has yetbeen discovered. The allozyme diversity andRAPD variation of Menziesia pentandrawere eight- and threefold greater than those ofM. goyozanensis, respectively, althoughwe examined only five populations representinga part of the distribution of M.pentandra. Because the environment surroundingthe population of M. goyoznaensis seemsto be stable at present, it is not necessary toplan an urgent ecological conservation practicefor the species. However, considering that thehabitat of M. goyozanensis is very smalland large vegetative disturbances occasionallyoccur in cool temperate forests, in which thespecies dwells, ex situ conservation isessential.     

Patterns of allozyme variation in relation to population size of the threatened plantMegaleranthis saniculifolia (Ranunculaceae) in Korea

Patterns of variation at 27 allozyme loci were investigated in the endangered endemic plantMegaleranthis saniculifolia. Levels of allozyme variation (A = 1.47,P = 40%,He = 0.088) were also compared with other endemic plant species. Genetic divergence between populations was very high (G ^st = 0.271 ), with moderate to high interpopulation differentiation, which probably arose through historical bottlenecks in a landscape of habitat fragmentation and/or human influence. The percentage of polymorphic loci, heterozygosity, and mean number of alleles per locus were positively related to population size, probably due to the stochastic loss of rare alleles in the smaller populations. Individuals in the small and marginal populations (TB, KD, and CJ) showed higher proportions of fixed loci. These ecologically marginal populations were typically more distant from the nearest neighboring population and were more genetically distinct from one another. The genetic structure of the current population ofM. saniculifolia is probably the result of local extinctions of intervening populations. This, in turn, is due to the Pleistocene climatic change and increased habitat destruction. A positive association appears to exist between genetic diversity and population size. Although these small population sizes are more sensitive to stochastic events, securing a certain number of individuals from the three larger populations (SB, JB, and TG) could be accomplished as part of a conservation strategy. In addition, it is important to prioritize populations in different regions in order to limit population declines caused by large-scale environmental catastrophes.     

Genetic diversity and structure of the endangered lady's slipper orchid Cypripedium japonicum Thunb. (Orchidaceae) in Japan

Cypripedium japonicum Thunb. (Orchidaceae), once a common perennial herb, is now designated as endangered throughout most of its distribution due to habitat destruction and fragmentation, and the impacts of horticultural collection. We investigated the genetic characteristics of this species for conservation purposes, using microsatellite markers to examine the genetic diversity and structure of 15 native and 5 ex situ populations in Japan. The results imply that although allelic variation is low in Japanese C. japonicum, sexual reproduction by seed, as well as clonal propagation, may occur in some populations. Both native and ex situ populations were found to be genetically differentiated, indicating that some populations may have experienced recent population declines, genetic fragmentation, or bottlenecks. The degree of genetic drift from the putative ancestral population, inferred through STRUCTURE analysis, was more pronounced in northern populations than in southern populations. Some of the ex situ conserved populations exhibited a low degree of differentiation from ancestral native populations. Our results imply that conservation of C. japonicum in Japan is best supported by maintaining individual populations and their unique genetic characteristics.