濒危灌木长叶红砂遗传多样性的RAPD分析

应用RAPD分子标记对濒危灌木长叶红砂(Reaumuria trigyna)种群遗传多样性进行了分析.应用18条随机引物对长叶红砂5个种群的95个个体进行扩增,检测到118个位点,其中多态位点105个.结果表明:长叶红砂种群的多态位点比率(P)为88.98%,显示出长叶红砂种群存在较高的遗传多样性.Shannon多样性指数(0.4966)、Nei基因多样性指数(0.3303)和基因分化系数(Gst=0.1425)的分析结果显示,长叶红砂种群遗传变异大多存在于种群内,种群间的遗传分化占14.25%.聚类分析表明,长叶红砂种群遗传距离与地理距离之间无直接相关关系.遗传多样性水平与物种特性和所处不同群落有关,濒危植物并不一定表现为遗传变异水平的降低.     

Volatile leaf oil diversity in the narrow range endemic Eucalyptus argutifolia (Myrtaceae) and its widespread congener Eucalyptus obtusiflora

Variation in the qualitative composition of volatile leaf oil was examined in the rare Eucalyptus argutifolia and its widespread congener Eucalyptus obtusiflora . The results revealed that, consistent with the pattern seen with allozymes, E. argutifolia had less variation within populations than E. obtusiflora . Total leaf oil diversity was also significantly lower in the rare species. As found with allozymes, most leaf oil diversity was within populations, but there was also a significant proportion of the variation between populations (25.2% and 27.3% for E. argutifolia and E. obtusiflora , respectively). There were significant associations between phenotypic distance based on leaf oils and geographical distance and between phenotypic and genetic distance across all populations, but these associations were not evident within species. Factors leading to reduced variation in E. argutifolia appear to affect all types of variation, but the relationships between different types of variation within the species are less apparent.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 738–745.     

Conservation of genetic diversity in the endangered plant Eriogonum ovalifolium var. vineum (Polygonaceae)

The purpose of his research was to describethe organization of genetic variation in thefederally endangered plant taxon Eriogonumovalifolium var. vineum using allozymes. Such information can help prioritize sites andmanagement choices for capturing andmaintaining genetic variation and can reducethe number of populations necessary to committo conservation, thus reducing costs andconflicts with competing land uses. Information on genetic diversity patterns alsoprovides insight into evolutionary anddemographic history of a taxon which canprovide means to assess future risk of erosionof diversity. Similar to other Eriogonumovalifolium varieties, E. ovalifoliumvar. vineum is diverse atboth the taxon (proportion of polymorphic loci[P] = 0.55 and alleles per locus [A] =5.45 [SD = 2.5]) and population (P = 0.56 [SD =0.11] and A = 2.68 [SD = 0.35]) levels. Gene diversity (H _e) withinpopulations averaged 0.19 (SD = 0.03). Wefound some evidence for limited clonalreproduction within populations. Populationswere moderately differentiated from one another(_S = 0.14) and showed moderatedeviations from Hardy-Weinberg equilibriumwithin populations (f = 0.14 and F =0.19). Mean Nei's genetic distance (D)among all pairs of populations was 0.02. Populations with high levels of multiplemeasures of genetic diversity, high levels ofdifferentiation, and low levels of apparentinbreeding are suggested as conservationpriorities.     

遗传多样性概述

遗传多样性作为生物多样性的重要组成部分,是物种多样性、生态系统多样性和景观多样性的基础。随着研究方法和实验技术的发展,遗传多样性研究从形态学水平、细胞学（染色体）水平、生理生化水平逐渐发展到分子水平。形态标记、细胞学标记、等位酶分析、DNA多态性分析等方法,为我们研究遗传多样性提供了有效的工具。特别是DNA多态性分析是一种更为直接而有效的方法。     

Assessment of genetic diversity in Coscinium fenestratum

Random amplified polymorphic DNA (RAPD) markers were used to assess the genetic diversity of 14 individuals belonging to 7 populations of Coscinium fenestratum (Gaertn.) Colebr. (Menispermaceae). 18 decamer primers used for the analysis generated 99 scorable bands of which 79 were found to be polymorphic. Coefficient of similarity ranged from 0.6604 to 0.9809. Variation within population was slightly higher than between populations. Similarity between individuals within and between populations was found. Dendrogram was obtained by using unwieghed pair-group method analysis (UPGMA). Distinct accession also exhibited higher percentage of medicinally active compound.     

Allozyme evidence for genetic autopolyploidy and high genetic diversity in tetraploid cranberry, Vaccinium oxycoccos (Ericaceae)

Polyploidy has been important in the evolution of angiosperms and may significantly affect population genetic diversity and structure. Nineteen isoenzyme loci were studied in diploid and tetraploid populations of Vaccinium oxycoccos (Ericaceae), and the results are compared with data previously reported for the related V. macrocarpon. Diploid V. oxycoccos and V. macrocarpon were readily discriminated based on their allozymic variation. No evidence for fixed heterozygosity was found in tetraploid V. oxycoccos. In contrast, all polymorphic loci exhibited both balanced and unbalanced heterozygotes, with some individuals exhibiting a pattern consistent with the presence of three alleles. These results support an autopolyploid origin for tetraploid V. oxycoccos. However, tetraploid V. oxycoccos possessed a suite of alleles not found in diploid V. oxycoccos; half of these alleles were shared with V. macrocarpon. This suggests that autotetraploid V. oxycoccos may have undergone hybridization with V. macrocarpon or that the autotetraploid retained the genetic variation present in an ancestral diploid species. Following theoretical expectations, proportion of polymorphic loci, mean number of alleles, and observed heterozygosity were significantly higher for the autotetraploid than for the diploid. Mean inbreeding (F(IS)) was similar for diploid and tetraploid V. oxycoccos. The latter exhibited population differentiation (F(ST)) exceeding both diploid species.     

西鄂尔多斯特有种四合木种群遗传多样性及遗传分化研究

利用垂直板聚丙烯酰胺凝胶电泳技术,对西鄂尔多斯高原特有种四合木（Tetraena mogolica）4个种群遗传多样性和遗传分化进行了初步研究。电泳结果表明,四合木在种和种群水平维持较高的遗传多样性,1多态位点百分率P＝60％,等位基因平均数A＝1.6,平均期望杂合度He=0.245。4个种群之间遗传分化很小,基因分化系数GST只有0.052,明不同于其它濒危物种。四合木种较高的遗传多样性和极低的种群间分化,说明不同的种群可能有共同的起源,随机遗传漂变和近交衰退不是影响遗传多样性的主要过程。     

濒危特有种掌叶木的微卫星遗传多样性研究

掌叶木居群具有较丰富的遗传多样性,该研究利用9对微卫星(SSR)分子标记揭示了掌叶木(Handeliodendron bodinieri)的遗传多样性。结果表明:观测等位基因数(Na)平均为3.903,有效等位基因数(Ne)平均为2.545,期望杂合度(He)平均为0.521,Shannon’s多态性信息指数(I)为0.962,PIC平均值为0.465。掌叶木的自然分布居群有相对较高的遗传多样性,但由于人为破坏等因素导致该群体濒危,而濒危并不是因为遗传多样性降低而造成的。居群间的遗传分化为掌叶木8个居群间的遗传一致度为(GI=0.849~0.970),遗传距离为(GD=0.032~0.164)。基于Nei’s遗传距离用UPGMA法对掌叶木居群进行聚类,Nei’s的基因分化系数为(G_(st))为0.027,平均Nei标准遗传分化系数(G'_(st)N)为0.031,平均Herick’s标准遗传分化系数(G'_(st)H)为0.064,基因流(N_m)为3.368。AMOVA分析结果表明:掌叶木居群间变异占3%,居群内变异占97%,居群内的遗传分化大于居群间的分化。利用Mantel检测发现,居群间的遗传距离与地理距离显著正相关(r=0.299,P0.05)。该研究结果为掌叶木生物多样性和资源保护与利用提供了更充分的科学依据。     

Extensive clonality in the endangered shrub Haloragodendron lucasii (Haloragaceae) revealed by allozymes and RAPDs

The occurrence of clonality in threatened plants can have important implications for their conservation. In this study, allozymes and RAPDs were used to determine the extent of clonality in the endangered shrub Haloragodendron lucasii (Haloragaceae), which is known from only four sites within an 8 km range. Allozyme markers identified only six multilocus genotypes among the 53 ramets sampled across the four sites, although a total of 54 different genotypes were possible with the three polymorphic allozyme loci detected. The polymorphic bands detected in the RAPD analysis were capable of producing 2⁴⁶ genotypes, but again only six multilocus genotypes were delineated. The allozyme and RAPD data were congruent at three of the four sites. At the fourth site two genotypes were detected by each marker; however, once combined, three multilocus genotypes were observed. The probabilities that the observed number of replicates of each combined allozyme and RAPD genotype could be generated by sexual reproduction were less than 10^–18, leaving little doubt that clonality is the explanation for the observed patterns of genotypes. The genetic conclusions are supported by root excavations which show potential for vegetative reproduction and the observation of no sexual reproduction in the species. The recognition of extensive clonality in H. lucasii has had immediate implications for the conservation management of the species and resulted in changes to the management priorities for the species. Thus it is clear that appropriate genetic studies can play an important role in the management of threatened species.     

The genetic structure of the steppe plant Iris aphylla L. at the northern limit of its geographical range

Genetic diversity of Iris aphylla was examined in seven Polish populations located at the northern limit of the geographical range of the species. Using RAPD analysis, 491 DNA fragments were amplified, all of which were polymorphic. The peripheral populations were characterized by a low number of unique bands (one to three per population) and moderate values of genetic diversity parameters (percent of polymorphic loci = 30.6%, Shannon's index = 0.149 and Nei's gene diversity = 0.097, on average), which can be explained as being due to the recent expansion of the European biota and the extremely low frequency or absence of seed set and of juveniles, as well as limited pollen and seed dispersal. There was also no significant correlation between ecological properties (population size; number of flowering ramets and fruits) and genetic diversity parameters. Despite this fact, more fixed loci were noted in the smallest populations than were in the largest ones. All populations were significantly differentiated and geographically structured. Analysis of molecular variance between regions showed a low and nonsignificant value, but relatively high molecular variation was partitioned among and within populations. Only one group of south-eastern populations was structured according to isolation by distance pattern and neighbour-joining cluster analysis. © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 152 , 245–255.     

Genetic diversity in Isoetes yunguiensis, a rare and endangered endemic fern in China

Isoetes yunguiensis is an endangered and endemic fern in China. Field survey indicated that only one population and no more than 50 individuals occur in the wild. The genetic variation of 46 individuals from the population remaining at Pingba (Guizhou Province, China) was assessed by Random Amplified Polymorphic DNA (RAPD) finger-printing. Twelve primers were screened from sixty ten-bp arbitrary primers, and a total of 95 DNA fragments were scored. Of these, 62.1% were polymorphic loci, which indicated that high level genetic variation existed in the natural population. The accumulation of genetic variation in the history of the taxon and the apparent minimal reduction effect on genetic diversity following destruction of habitat might be responsible for the high level genetic diversity presently remaining in the I. yunguiensis population. However, with the continuing decrease of population size, the genetic diversity will gradually be lost. We suggest that the materials from the extant population should be used for re-establishment of the populations. Translated from Journal of Wuhan University (Natural Sciences Edition), 2005, 51(6): 767–770 [译自: 武汉大学学报 (理学版)]     

Random amplified polymorphic DNA (RAPD) markers reveal less genetic variation in the endangered plant Cerastiumfischerianum var. molle than in the widespread conspecific C. fischerianum var. fischerianum (Caryophyllaceae)

Random amplified polymorphic DNA (RAPD) analysis was used to assess the level of genetic diversity in an endangered plant Cerastiumfischerianum var. molle and the widespread conspecific C. fischerianum var. fischerianum. At both the variety and the population level, C. fischerianum var. molle had a lower level of RAPD variation than var. fischerianum. Analysis of molecular variance ( AMOVA ) showed that most of the genetic diversity exists among populations for both varieties. Genetic differentiation among populations probably resulted from the highly selfing habits of both varieties. To conserve C. fischerianum var. molle genetically, preservation of only a part of the population is insufficient.     

Genetic diversity and species identification in the endangered white abalone (<Emphasis Type="Italic">Haliotis sorenseni</Emphasis>)

In 2001, the white abalone Haliotis sorenseni became the first marine invertebrate in United States waters to receive federal protection as an endangered species. Prior to the endangered species listing, 20 abalone were collected as potential broodstock for a captive rearing program. Using DNA from these animals, we have developed genetic markers, including five nuclear microsatellite loci and partial sequences of one nuclear (VERL) and two mitochondrial (COI and CytB) genes, to assess genetic variability in the species, aid in species identification, and potentially track the success of future outplanting of captive-reared animals. All five microsatellite loci were polymorphic and followed expectations of simple Mendelian inheritance in laboratory crosses. Each of the wild-caught adult abalone exhibited a unique composite microsatellite genotype, suggesting that significant genetic variation remains in natural populations. A combination of nuclear and mitochondrial gene sequencing demonstrated that one of the original wild-caught animals was, in fact, not a white abalone, but H. kamtschatkana (possibly subspecies assimilis). Similarly, another animal of uncertain identity accidentally collected by dredging was also shown to be H. kamtschatkana. Inclusion of these two animals as broodstock could have resulted in unintentional hybridizations detrimental to the white abalone recovery program. Molecular genetic identifications will be useful both in preventing broodstock contamination and as markers for future restocking operations.     

Genetic diversity in the threatened insular endemic plantAster asa-grayi (Asteraceae)

Genetic diversity in an insular endemic plantAster asa-grayi was examined using enzyme electrophoresis. Distribution ofA. asa-grayi is restricted to only four subtropical islands of Japan, and this species is listed as vulnerable to extinction in the Red Data Book of Japanese wild plants. A total of 161 individuals were sampled from five populations on four islands. Genetic diversity values at the population level were very low, compared to other plant species with a similar life history. Genetic variability at the species level is comparable to the mean value of endemic species. Genetic differentiation among populations is extremely high (G_ST= 0.71), indicating that the gene flow among populations is highly impeded, and pollen and seed dispersal is limited due to the pollinators and the seed morphology. This is because the four islands are geographically isolated. Fixation indices suggested that most populations do not randomly cross. To conserve the genetic diversity of the species, artificial crossings among different island populations are necessary.     

缙云山特有植物缙云黄芩的遗传多样性研究

采用聚丙烯酰胺凝胶垂直平板电泳检测了重庆缙云山特有植物缙云黄芩7个居群70个个体的过氧化物酶、细胞色素氧化酶、超氧化物歧化酶、淀粉酶和酯酶5种等位酶,获得20个基因位点的资料,由此分析了其遗传分化水平。结果表明：缙云黄芩遗传多样性水平较高,多态位点比率P为45．7％,平均每个多态位点的等位基因数目A为1．46,平均预期杂合度He为0．205,平均观察杂合度Ho为0．352。缙云黄芩居群间遗传分化水平较高,Gst=0．401,居群间遗传一致度和遗传距离的平均值分别为0．741和0．300。缙云黄芩遗传多样性的40．1％来源于居群之间的基因差异,59．9％属于居群内的遗传分化,因而对缙云黄芩遗传多样性的保护,应保护其较多的居群。     

Relationships and genetic diversity of endemic Elaphoglossum from St Helena

Allozyme polymorphism was used 1) to investigate the relationships of three threatened species of Elaphoglossum from St Helena, E. nervosum, E. bifurcatum and E. dimorphum, and 2) to estimate levels of genetic diversity and its partitioning among populations. Despite showing morphological and ecological variation, the three species are closely related with high genetic identities. Evidence from one enzyme locus (Mdh-1) suggests that E. dimorphum is of hybrid origin involving E. nervosum and E. bifurcatum. Levels of genetic diversity were low in the three species, but comparable with other insular endemic angiosperms. Populations of E. nervosum and E. bifurcatum showed significant genetic differentiation, which should be taken into account in any conservation programme.     

四川西部濒危植物桃儿七遗传多样性的RAPD分析

桃儿七是一种具有重要药用价值的珍稀濒危植物。采用RAPD分子标记技术,对在四川西部地区的桃儿七7个自然种群的遗传多样性水平和遗传结构进行了分析。用12个RAPD引物对7个种群共140个样品进行了扩增,共得到111条清晰带,其中32条具有多态性,在物种水平上多态位点百分率(PPB)为28.83%,在种群内的多态位点百分率变动幅度为4.50%至16.22%。同其它一些濒危植物相比,桃儿七种群具有较低的遗传多样性(He=0.0622,Ho=0.0987)。7个自然种群间出现了很强的遗传分化,分化指数接近70%。种群间的基因流低(Nm=0.2240)。造成上述结果的可能原因是与桃儿七的繁育方式和有限的基因流等因素有关。应将遗传多样性相对较高的松潘县牟尼沟种群作为原位保护的核心种群进行保护,尽量保护所有现有种群。     

Analysis of the genetic diversity and relationships among and within species of Hippophae (Elaeagnaceae) based on RAPD markers

RAPD markers were used to assess the genetic diversity and inter- and intra-specific relationships of the genus Hippophae L. and to study the correlation between genetic distances and geographic distances among populations of H. rhamnoides ssp. sinensis. The results analyzed by the percentage of polymorphic loci and Shannon information index indicated that a high level of genetic diversity existed both among and within species of the genus Hippophae. In the UPGMA dendrogram, the species or subspecies were clustered into two main groups but not strictly grouped according to sect. Hippophae and sect. Gyantsensis Lian. The multiple regression analysis and Mantel test both indicated a significant correlation between genetic distance and altitude distance among populations of H. rhamnoides ssp. sinensis, and the cluster analysis suggested that the genetic variation among populations of H. rhamnoides ssp. sinensis was linked to their monophyletic origin. Moreover, some degree of genetic differentiation was found among samples collected at different times.     

Development of microsatellite markers for the endangered semi‐shrub Chimaphila umbellata (Ericaceae)

Based on the DNA sequencing reads obtained using 454 pyrosequencing, primers amplifying 16 microsatellite loci were developed for the endangered semi‐shrub Chimaphila umbellata, which occurs sporadically in the Japanese Archipelago. These 16 loci were polymorphic in the populations sampled from the Hokkaido and Tohoku Districts; the mean number of alleles was 3.31 and 3.44, and the mean expected heterozygosity was 0.42 and 0.44, respectively. These loci were not linked to each other and contained no null alleles. Amplification using these primers was also tested in the congeneric species C. japonica, but only three of them successfully amplified DNA of the species. These markers will be used to examine genetic diversity and genetic differentiation in populations of C. umbellata.     

濒危植物毛瓣金花茶与其同属广布种茶光合特性的比较

毛瓣金花茶为山茶科山茶属植物,该研究分别对野生种群和栽培种群的毛瓣金花茶及其同属广布种茶的光合特性进行了测定及差异比较。结果表明:在野生和栽培环境下,毛瓣金花茶的光补偿点(LCP)(分别为1.17和3.87μmol·m-2·s-1)和光饱和点(LSP)(分别为395.8和423.6μmol·m-2·s-1)均较低,最大净光合速率(Pmax)(分别为4.25和3.91μmol·m-2·s-1)较小,是典型的阴生植物;而茶的LCP(分别为6.57和9.09μmol·m-2·s-1)较低,LSP(分别为765.0和809.6μmol·m-2·s-1)较高,Pmax(分别为9.37和9.75μmol·m-2·s-1)较大,为耐荫植物。野生和栽培的毛瓣金花茶的Pmax、表观量子效率(AQY)、最大羧化速率(Vcmax)、最大电子传递速率(Jmax)和潜在最大净光合速率(Pmax)均显著低于茶(P0.05),这表明毛瓣金花茶的光合能力和CO2利用能力都比茶要弱。栽培的毛瓣金花茶叶片的Chla、Chlb、Chl(a+b)含量与茶相比无显著差异(P0.05),表明毛瓣金花茶较低的光合能力与其叶绿素含量无关。野生和栽培的毛瓣金花茶的叶面积与茶相比无显著差异(P0.05),而比叶重则显著高于茶(P0.05),与茶相比,毛瓣金花茶对光强的适应范围狭窄,光合能力和CO2利用能力低下,这可能是其分布狭窄的重要生理原因。