RAPD标记鉴别中华绒螯蟹种群的初步研究

用２００个随机引物对中华绒螯蟹（Ｅｒｉｏｃｈｅｉｒｓｉｎｅｎｓｉｓ）辽河种群、长江种群和瓯江种群进行了ＲＡＰＤ遗传标记鉴别研究,其中引物ＨＸ０１和ＨＸ０２检测到瓯江种群和辽河种群所有样品共有的ＨＸ０１０．４和ＨＸ０２０．７扩增片段,Ｍｒ分别为０．４ｋｂ和０．７ｋｂ．这两个扩增片段在模板ＤＮＡ浓度作上下５０％的改变、或在镁离子和引物浓度分别作上下２５％的改变时仍稳定出现,而长江种群样品的ＰＣＲ反应中无这两个扩增片段出现,因而可作为长江种群的鉴别标记．用上述标记检查一些人工养殖的河蟹,发现一些苗种场的蟹苗种质严重混杂．未发现可以区分瓯江种群和辽河种群的标记     

威海荣成鹿角菜野生种群遗传多样性及资源保护策略

利用随机扩增多态性DNA（RAPD）方法对威海荣成地区鹿角菜（Silvetiasiliquosa）野生种群共30个个体的遗传多样性水平及遗传结构进行研究。结果表明,利用21条随机引物共检测到112个多态性位点,多态性位点百分率为93．75％,Nei基因多样性指数为0．3515±0．1352,Shannon多样性指数为0．5046±0．1126。通过聚类分析而划为一类的3个亚种群间的遗传分化指数为0．1349～0．2189,基因流为1．7793。研究结果表明威海荣成地区的鹿角菜种群遗传多样性较高,种群内存在较为明显的遗传分化,种群遗传资源不受遗传漂变的影响。针对当地鹿角菜遗传资源的现状,应加强现存种群的就地保护,恢复种群规模;进行取样养殖保护时则需避免因近交而造成种群资源退化。     

RAPD标记鉴别中华绒螯解种群的初步研究

用２００个随机引物对中华绒螯蟹辽河种群,长江种群和瓯江种进行了ＲＡＰＤ遗传标记鉴别研究,其中引物ＨＸＯ１和ＨＸＯ２检测与瓯江种群和辽河种群所有样品共有的ＨＸＯ１－０．４和ＨＸ０２－０．７扩增片段,     

山西高原油松种群遗传多样性

用酸性聚丙烯酰胺凝胶电泳（A-PAGE）技术，分析了山西高原9个油松种群在醇溶蛋白水平上的遗传多样性。135份材料共分离出23条带，其中3条为共有带，多态性高达86．95％。全部材料共出现53种带型，9个不同油松种群的带型有差异，同一种群不同个体的带型也有所不同，说明山西高原的油松在遗传上已产生一定程度分化，在醇溶蛋白水平上呈现出遗传多态性。从供试材料的带型计算出油松遗传分化系数为0．1547。即在种群间的变异占总变异的15．47％，种群内变异为84．53％，大部分的遗传变异存在于种群内，但种群间的分化程度在松属树种中也属于较高水平。根据23个多态位点计算遗传相似系数和遗传距离，进行聚类分析，将山西高原9个油松种群聚为3个类群。     

大贵寺国家森林公园野生青檀居群的遗传多样性

利用ISSR分子标记技术对湖北省大贵寺国家森林公园野生青檀（Pteroceltis tatarinowii Maxim．）居群进行遗传多样性分析。用11条ISSR引物对6个青檀居群64个样品进行扩增,扩增片段长度在200bp至3000bp之间,检测到179个位点,其中多态位点165个,多态位点百分率（P）为92．14％。青檀在物种水平上的Nei基因多样性指数（胁）和Shannon信息指数∽分别为0．273和0．423,在居群水平上分别为0．225和0．343,显示出青檀有着较高的遗传多样性。居群间的遗传分化系数（Gsr）为0．192,表明青檀居群间存在着较大的遗传分化;UPGMA聚类分析表明遗传距离与海拔差距有一定的相关性,相邻海拔的青檀居群间遗传距离较近。不同海拔导致的异质生境可能是影响青檀居群遗传分化的主要原因。     

黄鳝野生群体和养殖群体遗传结构的RAPD分析

采用RAPD标记技术对黄鳝野生群体和养殖群体遗传结构进行初步研究.用24个随机引物在野生群体和养殖群体中分别扩增出259和251条DNA片段,其中多态性片段数分别为116和92条,多态性片段的比例分别为44.79%和36.65%,平均杂合度分别为0.2155和0.1908,遗传相似率分别为0.9053和0.9583,群体间遗传相似率为0.8863.而Shannon遗传多样性指数表明,两群体中的遗传变异有81.51%来自群体内,只有18.49%来自群体间.野生群体的多态性位点比例和杂合度明显高于养殖群体,说明黄鳝野生种质资源状况较好,应加以合理保护.与野生群体相比,养殖群体的杂合度和遗传多样性有所下降,提示在黄鳝的人工繁殖过程中应引进标记辅助选择等技术手段.图1表3参24     

游蛇科（Colubridae）10种的随机扩增多态DNA研究

用随机扩增多态ＤＮＡ（ＲＡＰＤ）技术对游蛇科１０种蛇基因组ＤＮＡ的多态性进行了分子遗传标记的研究，所得数据经聚类分析结果提示，１．蛇类种内个体间在着遗传多样性妈个体间的差异，随机扩增多态ＤＮＡ片段共享度的分析表明，种间差异显著大于种内个体间的差异，说明在研究种间系统演化关系时，可以用随机取样的个体代表一个种作种间的比较．２．锦蛇属是一高度分化的大属，本研究中的５种锦蛇间，玉斑锦蛇和红点锦蛇关系的较     

中国茸鹿品种(品系)的随机扩增多态DNA(RAPD)研究

采用随机扩增多态ＤＮＡ技术研究了５个茸鹿品种（品系）７５个个体的遗传变异关系。在所使用的４１个引物中,有４０个引物扩增出多态谱带,共检测到３９５条扩增片段,其中２５９条（６５．６％）出现变异,用Ｓｈａｋｎｎｏｎ指数计算了５个品种（品系）的遗传变异及其遗传变异在群体内和群体间的分布。利用Ｎｅｉ氏片段共享度计算了７５个个体间的遗传距离,用ＵＰＧＭＡ聚类法构建了７５个个体的系统发育树状图,反映５个茸鹿品     

金花茶遗传多样性和遗传结构AFLP分析

利用扩增片段长度多态性(amplified fragment length polymorphism,AFLP)分子标记技术,对金花茶(Camellia nitidissima) 4个天然种群的遗传多样性和遗传结构进行研究,为金花茶资源的有效保护和高效利用提供科学依据。结果表明,8对引物共扩增出1 619个DNA位点,其中1 473个位点具多态性,多态性位点百分率为90. 98%。金花茶物种水平基因多样性指数和Shannon信息指数为0. 146和0. 246,种群水平基因多样性指数和Shannon信息指数为0. 131和0. 215; 4个金花茶种群的多态位点百分率、基因多样性指数和Shannon信息指数变化趋势一致,其中多态位点百分率介于56. 83%～72. 11%之间,基因多样性指数介于0. 122～0. 138之间,Shannon信息指数介于0. 197～0. 228之间。金花茶种群间的遗传分化系数介于0. 139～0. 289之间,遗传变异主要存在于种群内。分子变异分析(AMOVA)结果表明,金花茶种群内遗传变异为81. 52%,种群间遗传变异为18. 48%,遗传变异主要存在于种群内。非加权组平均法(UPGMA)聚类结果表明,种群间遗传变异与其地理距离无明显相关性。应加强现有金花茶种群的就地保护,促进种群自然更新,同时加强迁地保护。     

游蛇科（Colubridae）10种蛇的随机扩增多态DNA研究

用随机扩增多态ＤＮＡ（ＲＡＰＤ）技术对游蛇科１０种蛇基因组ＤＮＡ的多态性进行了分子遗传标记的研究．所得数据经聚类分析结果提示：１．蛇类种内个体间存在着遗传多样性即个体间的差异，随机扩增多态ＤＮＡ片段共享度的分析表明，种间差异显著大于种内个体间的差异，说明在研究种间系统演化关系时．可以用随机取样的个体代表一个种作种间比较．２．锦蛇属是一高度分化的大属，本研究中的５种锦蛇间．玉斑锦蛇和红点锦蛇关系较近．可以井为１组．另３种归为１组还是分为３个不同的组尚难定论．３．游蛇科６个属之间．锦蛇属、乌梢蛇属和鼠蛇属３属间亲缘关系较近．Ｒｈａｂｏｐｈｉｓ和Ｓｉｎｏｎａｔｒｉｘ与链蛇属较近．它们可能代表该科中较原始的类群．     

Analysis of global and local population stratification of finless porpoises <Emphasis Type="Italic">Neophocaena phocaenoides</Emphasis> in Chinese waters

The existence of three distinct populations is widely accepted for the finless porpoise (Neophocaena phocaenoides) in Chinese waters: the Yellow Sea, Yangtze River, and South China Sea populations. Here, we use nine species-specific microsatellite loci, the complete mitochondrial DNA control region (912 bp), and the complete mitochondrial cytochrome b gene (1,140 bp) to further investigate potential population stratification in the Yellow Sea using 147 finless porpoise samples from the Bohai Sea and adjacent northern Yellow Sea, two regions that were largely underrepresented in previous genetic studies. Our F-statistics analyses confirm the previously described three populations, but further demonstrate significant genetic differentiation between the [Bohai + northern Yellow] Sea and the southern Yellow Sea. On the other hand, median-joining network analyses do not exhibit well-differentiated haplotype groups among different geographic populations, suggesting the existence of shared ancestral haplotypes. Levels of microsatellite diversity are moderate to high (mean H _E = 0.794) among the 147 [Bohai + northern Yellow] Sea finless porpoises and no recent bottleneck was detected, whereas mtDNA control region and cytochrome b gene diversity is low to moderate. The microsatellite genotypic and mtDNA haplotypic data also confirm the presence of mother-calf pairs in single-net bycatch cases. The results presented here highlight the necessity to treat the [Bohai + northern Yellow] Sea population (highly impacted by anthropogenic threats) as a separate Management Unit.     

Genetic Diversity and Tests of the Hybrid Origin of the Endangered Yellow Larkspur

Abstract: Delphinium luteum ( Ranunculaceae), an endangered larkspur, is restricted to two wild populations near Bodega Bay, California. The total number of individuals in these two populations is estimated to be <100. We used allozyme and random amplified polymorphic DNA ( RAPD) markers to (1) assess levels and patterns of genetic diversity in one wild population and two cultivated populations and (2) test the hypothesis that D. luteum is of hybrid origin between D. decorum and D. nudicaule . These data will be used to aid in developing a management plan to conserve the species. The wild population maintains high levels of genetic diversity. Genetic data indicate that both cultivated populations, especially the north Sonoma population, have several allozymes and RAPD markers not found in the wild population and could be used to establish new populations of D. luteum or to enhance the diversity and size of the wild population. The allozyme data did not reveal any fixed differences between D. decorum and D. nudicaule , although allele frequencies of the putative parental populations differed. At these loci, D. luteum resembled D. nudicaule more than D. decorum  . Many unique RAPD markers distinguish each of the three species. The diagnostic markers from populations of D. nudicaule and D. decorum were not additive in the putative hybrid, and these data indicate that D. luteum is not of recent hybrid origin. Conservation of the yellow larkspur should include strategies that use the cultivated populations of D. luteum , but hybridizing D. decorum and D. nudicaule to "recreate" D. luteum is not recommended.     

A_2型高粱细胞质雄性不育系与其保持系的胞质DNA和核DNA差异

以高粱细胞质雄性不育系A2 V4 (A)及其保持系V4 (B)的总DNA为模板 ,对 184个随机引物进行筛选 ,找到6个其RAPD扩增产物在A/B间存在稳定差异的引物 .将该 6个引物同时扩增A/B的总DNA、线粒体DNA(mtDNA)及叶绿体DNA(cpDNA) .以总DNA为模板时得到 12个扩增片段 ,以mtDNA为模板时得到 4个 ,以cpDNA为模板时得到 11个 .结果分析表明 ,在这些扩增片段中 ,有 7个仅仅出现在以胞质DNA为模板的扩增中 ,有 5个在以总DNA和胞质DNA为模板时同时出现 ,即认为这 12个片段来自胞质DNA .另有 7个片段 ,在以胞质DNA为模板时未出现 ,而是仅仅出现在以总DNA为模板的扩增中 ,认为是来自核DNA .来自核DNA的 7个扩增片段中 ,有 5个来自保持系 ,有 2个来自不育系 ,这表明 ,不育系与保持系在核DNA上存在差异 .对A/B核DNA在CMS中的重要性及研究对策进行了讨论 .图 2表 4参 19     

Application of random amplified polymorphic DNA (RAPD) markers to evaluate intraspecific genetic variation in red mullet (Mullus barbatus)

The random amplified polymorphic DNA (RAPD) technique was used to evaluate genetic affinities among eight red mullet (Mullus barbatus L., 1758) samples from the Mediterranean Sea. Twenty-nine random primers were used. Despite the variability which was found within samples, no specific RAPD marker for the discrimination of the populations was detected. The data analysis revealed that the genetic diversity among populations is positively related to their geographic distances. The results of this study indicate that the estimated intraspecific variation may be more␣pronounced with RAPD markers than with allozymes when the two approaches are applied on the same populations. Received: 7 November 1997 / Accepted: 30 May 1998     

利用RAPD标记分析卧龙自然保护区不同海拔沙棘种群的遗传变异

从1800m到3400m五个海拔连续取样,用RAPD分子标记研究了卧龙自然保护区中国沙棘种群的遗传结构和遗传变异.用11条寡核苷酸引物,扩增得到151个重复性好的位点,其中143个多态位点,多态率达94.7%.在5个沙棘种群中,总遗传多样性值(HT)为0.289,B种群内的遗传多样性值为0.315,这完全符合沙棘这种多年生、远交的木本植物高遗传变异的特性.5个种群内遗传多样性随海拔升高呈低—高—低变异趋势,在2200m海拔处的B种群遗传多样性达最大值0.315,3400m海拔处的E种群则表现最小,仅0.098.5个种群间的遗传分化值GST=0.406,也即是说有40.6%的遗传变异存在于种群间,59.4%存在种群内.1800m海拔处的A种群与其他种群的明显分离是造成种群间遗传分化大的原因.UPGMA聚类图和PCoA散点图分别进一步确证了5个种群间关系和所有个体间的关系.最后,经过Mantel检测,遗传距离与海拔表现了明显的相关性(r=0.646,P=0.011).     

RAPD分子标记在鉴定香樟优选株和普通株中的应用

从85个随机引物中选10个随机引物对8个香樟样品的DNA进行了扩增，应用RAPD技术区别6个优选株和2个普通株，共获得78个DNA片断，把这些DNA片断作为遗传位点用UPCMA法计算出各材料间的遗传相似性系数，并作了聚类分析．在D＝0．738处8个样品分为两大类，而且普通株同优选株的亲缘关系很近．图2表2参9     

Effects of Population Size, Mating System, and Evolutionary Origin on Genetic Diversity in Spiranthes sinensis and S. hongkongensis

Hong Kong once supported more than 109 species of wild orchids, of which approximately 30% were endemic. Most of the local wild orchids have now become rare or endangered. I conducted a comparative study of genetic diversity in two closely related terrestrial orchids, an allotetraploid, Spiranthes hongkongensis , and its diploid progenitor, S. sinensis , to assess the effects of the population bottleneck associated with the origin of the polyploid and to investigate the relationships between number of breeding individuals, mating system, and level of isozyme variation in their populations. Nearly complete genetic uniformity was observed both within and among populations of S. hongkongensis . In contrast, S. sinensis had high levels of genetic variation for all of the genetic parameters examined. Regression analysis of population size and several components of genetic diversity in S. sinensis revealed that, among various measures of within-population variation, the proportion of polymorphic loci ( P ) and average number of alleles per locus ( A ) or per polymorphic locus ( A _p ) were the most sensitive to population size ( R ² = 0.942, p = 0.001; R ² = 0.932, p = 0.002; and R ² = 0.923, p = 0.002 respectively). The highly negative correlation ( r = −0.999, p < 0.01) between population size and the mean frequency of private alleles in pairwise population comparisons, p (1), indicated that population size may also be used to predict the extent of population differentiation caused by random genetic drift. Conservation of genetic diversity in S. sinensis could be maximized by protecting several of both large and small populations, whereas fewer populations may be needed to achieve this goal for S. hongkongensis.     

Genetic structure and allozyme variation of sea bass (Dicentrarchus labrax and D. punctatus) in the Mediterranean Sea

This paper reports data on 28 allozyme loci in wild and artificially reared sea bass (Dicentrarchus labrax) samples, originating from either coastal lagoon or marine sites in the Mediterranean Sea. F _ST analysis (θ estimator) indicated strong genetic structuring among populations; around 34% of the overall genetic variation is due to interpopulation variation. Pairwise θ estimates showed that, on average, the degree of genetic structuring was much higher between marine populations than between samples from lagoons. Six polymorphic loci showed differences in allele frequencies between marine and lagoon samples. Multivariate analyses of individual allozymic profiles and of allele frequencies suggested that different arrays of genotypes prevail in lagoons compared to marine samples, particularly at those loci that, on the basis of previous acclimation experiments, had been implicated in adaptation to freshwater. On the other hand, variation at “neutral” allozyme loci reflects to a greater extent the geographic location of populations. Allozyme differentiation was also studied in a D. labrax population from the Portuguese coast. Average genetic distance between this population and the Mediterranean populations was quite high (Nei's D = 0.236) and calls into question the taxonomic status of the Portuguese population. Finally, genetic relationships between D. labrax and D. punctatus were evaluated. Average Nei's D was 0.648, revealing high genetic differentiation between the two species, even for two sympatric populations of these species in Egypt; thus gene flow was not indicated between species. Received: 24 October 1996 / Accepted: 27 November 1996