濒危植物三棱栎遗传多样性的RAPD分析

用随机扩增多态DNA (RAPD)标记对 5个三棱栎 (Trigonobalanusdoichangensis)居群共 99个个体进行遗传多样性和居群遗传结构分析。 16个引物共检测到 15 7个位点 ,其中多态位点 83个 ,占 5 2 87%。物种水平Shannon多样性指数I =0 2 4 31,Nei基因多样度h =0 15 95 ,种内总遗传变异量Ht=0 16 0 0 ,居群内遗传变异量Hs =0 0 74 9,居群间变异量大于居群内变异量 ,表明三棱栎的遗传变异主要存在于居群之间。与同科植物相比 ,三棱栎遗传多样性较低 ,遗传分化系数Gst =0 5 32 0 ,说明居群间的遗传变异占 5 3 2 0 % ,居群间已出现强烈的遗传分化。当地人的强烈活动造成的生境破碎化和居群隔离 ,以及三棱栎演化过程中的地史变化对其种群发展的影响等 ,可能是造成其居群间强烈的遗传分化和较低遗传多样性的原因。基于本研究结果 ,提出了三棱栎遗传多样性的保护策略。     

南方红豆杉的ISSR遗传多样性分析

采用ISSR标记技术对南方红豆杉迁地保护小种群及其衍生自然种群5个小斑块(小居群)的遗传多样性进行了分析.从90条引物中共筛选出10个多态性引物,获得ISSR谱带102条,其中多态性谱带74条,占72.54%.结果显示,南方红豆杉迁地保护小种群和迁地保护衍生自然种群的Nei's基因多样性指数(H)分别为0.220 9和0.254 8,Shannon信息指数(I)分别为0.322 4和0.376 2.5个小斑块居群的H分别为0.190 7、0.210 4、0.194 4、0.160 5和0.187 3,I分别为0.281 0、0.309 8、0.288 0、0.235 5和0.273 5.迁地保护衍生自然种群的遗传多样性高于迁地保护小种群,但各小生境下的的小斑块居群遗传多样性低于迁地保护小种群.聚类分析结果表明:南方红豆杉迁地保护各自然小居群间的遗传距离与这些居群的地理生境有关,而与地理距离并没有显著相关性.     

西藏特有植物砂生槐天然居群遗传多样性研究

采用等位酶淀粉凝胶电泳技术对西藏雅鲁藏布江中游砂生槐 (Sophoramoorcroftiana) 10个天然居群的遗传多样性进行了研究。 13个酶系统 2 4个酶位点 ( 46个等位基因 )的检测结果表明 ,砂生槐具较低的遗传变异水平。居群水平上的遗传多样性指标分别为 :多态位点百分率Pp =2 5 .0 %～ 37.5 % ,等位基因平均数Ap=1.3～ 1.7,平均期望杂合度Hep=0 .112～ 0 .16 9;种水平上的遗传多样性 (Ps=37.5 % ,As=1.9,Hes=0 .171)低于长寿命木本被子植物的平均值 (Ps=5 9.5 % ,As=2 .10 ,Hes=0 .183)。居群遗传结构的分析显示 ,10个居群中随机交配的偏差为FIS=- 0 .0 0 71,表明砂生槐在居群水平上存在轻微的杂合子过量现象 ,偏离了Hardy Weinberg平衡 ;FST=0 .1748,表明砂生槐是居群间分化较大的一类多年生木本植物 ,主要原因是环境恶化和人类活动干扰 (过度砍伐、放牧等 )导致其生境片断化 ,从而影响了居群间基因交流而造成基因流水平较低 (Nm =1.180 2 )。砂生槐高海拔居群H2 (谢通门 )、H31(江当 1)、H32 (江当 2 )、H5 (朗塞岭 )包含着绝大部分等位基因 ,显示了相对较高的遗传多样性水平 ,应加以保护和管理 ,作为砂生槐种质资源就地保存的基地。     

承德光秃山不同海拔油松居群遗传多样性

为了解光秃山不同海拔下天然油松居群之间的遗传差异及探求遗传多样性与土壤因子的关联,为油松种源筛选和管理提供参考,本研究运用ISSR技术,对位于河北承德辽河源光秃山4个不同海拔油松天然居群共118个植株个体的遗传多样性进行分析。13个引物共扩增出177条清晰的条带,种群多态位点百分比(PPL)为60.2775%,Nei’s基因多样性指数(h)为0.2171,Shannon信息指数(I)为0.3222;不同种群遗传变异水平随海拔差异呈规律性变化,表现为沿海拔升高而呈低-高-低的分布规律,其中1354～1274 m范围的遗传多样性水平最高;在物种水平上油松具有较高的遗传多样性(PPL=98.33%,h=0.38142,I=0.5550),种群间的遗传分化系数Gst=0.6562。利用AMOVA软件对遗传变异的等级剖分结果表明,种群间有显著的遗传分化,约2/5的遗传变异存在种群间,种群内占3/5。Pearson相关分析表明,油松居群内遗传多样性与海拔、土壤养分(有机质、速效磷、速效钾含量)之间存在显著或一定的相关关系。Mantel检验结果显示,油松居群遗传距离与海拔差距、土壤养分因子的分异存在一定相关性。以上结果表明不同海拔区域的生态因子、低基因流等对油松居群间的遗传分化影响较大。     

云南灯盏花遗传变异的RAPD分析

运用RAPD (randomamplifiedpolymorphicDNA)技术对云南灯盏花 (Erigeronbrevis capus) 6个居群进行分析 ,研究其遗传变异及遗传结构 ,用外类群紫菀对比分析其亲缘关系。随机挑选 16个引物在 6个居群中共产生 2 33条DNA片段 ,其中 192条带具有遗传多态性 ,约占 82 4 0 %。多态位点百分比PPB、等位基因数A、有效等位基因数Ae、Nei’s基因多样性指数H和Shannon多样性指数I在物种水平分别为 82 4 0 %、 1 82 4 0、 1 30 0 5、0 1896、 0 30 2 1;居群的平均值分别为 5 4 2 3%、 1 5 40 1、 1 2 6 91、 0 16 0 7、 0 2 4 6 0。灯盏花的遗传多样性较丰富 ,基因分化系数Gst值为 0 346 0 ,有 6 5 4 0 %的变异来自居群内 ,6个居群的遗传一致度较高 ,在 0 90 37～ 0 972 3之间 ,平均为 0 94 10。利用UPGMA对 6居群聚类分析 ,结果表明 :丘北居群 (YB)和文山居群 (YX)亲缘关系最近 ,小哨居群 (Y )和新平居群 (YA)次之 ,丘北居群 (YB)和腾冲居群 (ZA)亲缘关系最远。遗传关系与各居群地理分布区间的距离大致成正相关     

四种松毛虫不同地理种群遗传多样性的等位酶分析

【目的】采用等位酶电泳技术对中国松毛虫属Dendrolimus 4种共9个地理种群进行遗传多样性和遗传分化研究。【方法】对6种等位酶系统乳酸脱氢酶（LDH）、苹果酸脱氢酶（MDH）、苹果酸酶（ME）、乙醇脱氢酶（ADH）、甲酸脱氢酶（FDH）、谷氨酸脱氢酶（GDH）进行聚丙烯酰胺凝胶电泳分析。【结果】在4种松毛虫9个地理居群中共检测到10个基因位点,其中4个位点为多态位点,检测到17个等位基因; 种群总体水平多态位点比率P=40%,平均有效基因数A=1.700,平均期望杂合度He=0.151,种群平均遗传距离为0.001～0.285; 其中马尾松毛虫指名亚种Dendrolimu punctatus Walker 6个居群的遗传分化度Fst=0.265,基因流Nm=0.692。4种松毛虫之间遗传关系最近的是落叶松毛虫D. superans Butler和马尾松毛虫的地理亚种赤松毛虫D. punctatus spectabilis Butler,遗传关系最远的是落叶松毛虫D. superans Butler和云南松毛虫D. houi Lajonquiere。【结论】 马尾松毛虫居群间遗传分化程度较大,基因交流较少,遗传漂变已经成为导致该物种种群分化的主要原因之一;遗传距离与地理距离存在一定相关性。     

极濒危植物中华水韭休宁居群的遗传结构

采用超薄平板微型聚丙烯酰胺等电聚焦电泳方法对极濒危蕨类植物中华水韭(Isoetes sinensis)现存于安徽休宁的5个亚居群的等位酶多样性和遗传结构进行了研究。结果表明：中华水韭居群每位点平均等位基因数A=1．7,平均多态位点比率P=55．56％,平均预期杂合度He=0．201。居群中半数的多念位点表现为等位基因的“固定杂合”,5个亚居群的遗传多样性无显著差异,但都表现出严重偏离Hardy—Weinberg平衡的杂合子过昔;其遗传变异主要发生于亚居群内(94．27％),亚居群之间的遗传分化较小(Gsl=0．0573),亚居群间遗传一致度较高(I=0．960—0．999)。我们推断这可能是由于居群构建之初的奠基者效应或者原种群曾经历了较为严重的遗传瓶颈所导致;此外亚居群间便利的基因交流也可能起到了很大作用(Nm=4．5062)。取自休宁居群的75株样品由13个专一多位点基因型(18个位点等位酶基因型)组成,亚居群及个体间高度遗传均质。本文还就中华水韭休宁野生居群濒危的遗传因素进行了探讨,并提出了相应的保育策略。     

三峡库区特有种疏花水柏枝的保护遗传学研究

采用超薄平板微型聚丙烯酰胺等电聚焦电泳方法对三峡库区特有种疏花水柏枝 (Myricarialaxiflora)的 13个自然居群和 1个人工迁地保护居群的等位酶遗传变异进行了初步研究。检测了 5个酶系统 ,得到 13个等位酶位点 ,遗传多样性及其遗传结构分析结果表明 :疏花水柏枝具有较高水平的遗传多样性 ,平均多态位点比率P =78.7% ,每位点平均等位基因数A =1.8,平均预期杂合度He=0 .317,高于中国植物特有种的平均水平 ,且群体中杂合基因型个体偏多 ;其遗传变异主要发生于居群内 ( 84.86 % ) ,居群间又存在一定的遗传分化 (Gst=0 .15 14) ,居群间平均基因流Nm =1.40 1,居群间遗传距离为 0 .0 0 2～ 0 .176 ;UPGMA聚类分析显示疏花水柏枝在三峡湖北境内的白水河—泄滩一带分为上游和下游 2个居群组 ;武汉植物园迁地保护的混合居群基本保育了其遗传多样性总水平。在分析讨论疏花水柏枝遗传多样性与其繁育系统、生境及其起源进化的关系的基础上 ,探讨了疏花水柏枝濒危的主要原因 ,推断其应为第四纪冰期影响后的古孑遗种。最后 ,在评价迁地保护成果的基础上 ,提出了今后进一步保育的策略。     

孑遗植物水杉的遗传多样性研究

采用随机扩增多态DNA(RAPD)方法对孑遗植物水杉 (Metasequoiaglyptostroboides)的 8个居群 (包括 6个孑遗居群和 2个人工栽培居群 )的 48个个体进行了遗传多样性研究。用 16个随机引物共扩增出清晰谱带 5 7条。孑遗居群平均多态位点百分率为 38.6 %。各居群多态位点百分率差异显著 ( 10 .5 3%～ 33 .33 % ) :湖北利川的小河和桂花 2个居群较高 ,分别是 33 .33%和 31.5 8% ;湖南龙山洛塔居群较低 ,为 10 .5 3 %。人工栽培居群中 ,美国密西根州和湖北武汉 2个居群的多态位点百分率也较低 ,仅分别为 10 .5 3 %和 8.77%。用POPGENE 1.31版软件进行数据处理后结果如下 :孑遗居群的平均Nei’s基因多样性为 0 .130 5 ,Shannon’s遗传多样性信息指数为 0 .192 1,水杉孑遗居群遗传距离与地理距离的相关系数r =0 .2 346 6 (P =73 .2 7% ) ,基因分化系数Gst=0 .10 82 ,表明居群间有一定分化。用UPGMA方法聚类可知 ,2个人工居群与湖北利川居群优先聚类。鉴于目前水杉孑遗居群生境人为破坏严重、遗传多样性较低和原生母树分布分散 ,建议在湖北利川小河和桂花野生水杉最适生境建立水杉遗传多样性保护点 ,将现有的 2 5 0多株原生母树分别采种育苗定植于保护点 ,以达到最大限度地保存现有水杉遗传多样性的目的。     

黄土高原不同气象因子对中国沙棘亚居群遗传多样性的影响

利用ISSR分子标记技术,分析了甘肃省东部地区8个中国沙棘居群的遗传多样性和遗传分化,以及气象因子对它们的影响.共调查了8个居群的240个个体,利用11条引物扩增出165条带.扩增范围集中在300～1 500 bp,其中157个位点表现为多态,占95.76%,分子变异分析(AMOVA)显示,居群内遗传分化程度高,76.5%的变异存在于居群内.基因分化系数(Gst)为0.2418,居群间基因流为1.5675.说明保护沙棘种群的遗传资源应优先考虑种群内个体.经Mantel检验发现,8个居群的地理距离和遗传距离显著相关(r=0.65,P=0.002). 对气象因子与遗传多样性所作的回归分析显示, 中国沙棘开花期间的风速与其遗传多样性呈显著正相关.说明开花期风速和地理距离是沙棘种群遗传多样性的重要影响因子.     

广西石灰岩地区蜈蚣蕨居群的遗传多样性研究

采用等位酶分析方法 ,研究了广西石灰岩地区蜈蚣蕨居群的遗传多样性 ,分析了其空间变化趋势。检测了 8个酶系统 ,1 5个酶位点。分析结果表明 :广西石灰岩地区蜈蚣蕨居群遗传多样性程度较高 ,每个位点的等位基因平均数为 1 .6 7,多态位点为 5 7.78% ,平均期望杂合度为 0 .2 4 9。蜈蚣蕨居群的遗传组成在居群间有一定的差异 ,但差异的程度并不与空间距离成正比     

Genetic diversity and population structure of the serpentine endemic Calystegia collina (Convolvulaceae) in northern California

We used enzyme electrophoresis to evaluate genetic diversity in 32 populations of Calystegia collina, a clonal plant species endemic to serpentine outcrops in northern California (USA). Of 34 loci examined 56% were polymorphic, but on average only 17% were polymorphic within local populations. Neither the total number of alleles nor the number of multilocus genotypes differed significantly between populations in small vs. large serpentine outcrops. Genetic and geographic distances between populations were positively correlated, but this relationship was not significantly affected by the isolation of serpentine outcrops. Populations were highly differentiated (F(st) = 0.417) and little genetic variation was explained by geographic region or serpentine outcrop.Observed heterozygosity within populations almost always exceeded Hardy-Weinberg expectations. In many populations, all 30 sample ramets were uniformly heterozygous at one or more loci yet were genetically variable at other loci. These results imply that many C. collina populations originate from one or a few genetic founders, with little recruitment from seeds. Genetic variation within uniformly heterozygous populations must be the product of multiple, closely related founders or somatic mutations within the population. We conclude that vegetative reproduction, perhaps coupled with somatic mutation, helps maintain genetic diversity in these isolated but long-lived populations.     

Allozyme variation in endangered Castanea pumila var. pumila

Allozyme genetic variation in 12 populations of the endangered Castanea pumila var. pumila (Allegheny chinkapin), sampled across the natural range of the species in the United States, was evaluated using 11 loci from seven enzyme systems. At the species level, the percentage of polymorphic loci (Ps) was 72.7 %, the mean number of alleles per locus (As) was 1.9, the mean number of alleles per polymorphic locus (APs) was 2.3, the effective number of alleles per locus (Aes) was 1.5 and the genetic diversity (Hes) was 0.296. At the population level, Pp = 49.2 %, Ap = 1.5, Aep = 1.4, APp = 2.1 and Hep = 0.21. Most of the allozyme variation (70 %) in C. pumila var. pumila occurred within populations. Wright's gene flow rate [Nm(W)] was as low as 0.57. Population differentiation along the species range was not detected. Populations of C. pumila var. pumila in Florida had the most variable levels of genetic diversity, but populations in Virginia and Mississippi also showed high levels. Based on the results of this study, conservation management strategies are discussed.     

Allozyme variation of Cyclobalanopsis championii (Fagaceae), a narrowly distributed species in southern Taiwan

Allozyme genetic variability in five natural populations of Cyclobalanopsis championii (Fagaceae) in Taiwan was investigated using 12 loci from 9 enzyme systems. The average values of parameters describing within-population variation, expected heterozygosity (He = 0.151), the percentage of polymorphic loci per individual (P = 50%), the average number of alleles per locus (A = 1.7), effective number of alleles per locus (Ae = 1.25), and the average number of alleles per polymorphic loci (AP = 2.2) are comparable to those of other long-lived woody plants. The overall fixation index (Fis = 0.208) indicates a significant deficiency of heterozygotes at the population level. Allelic frequency deviation from Hardy-Weinberg equilibrium was found for different loci in different populations. An exact test for population differentiation using the Tools for Population Genetic Analyses program also indicates that allelic frequencies among populations are significantly different (P < .001). Among-population variation, Gst, accounted for 9.2% of the total heterozygosity. The population at Shouchia and the southernmost population Nanjenshan had higher inbreeding coefficients (0.177 and 0.153, respectively) than did the northern populations. Genetic drift is supported by the observations of the variance components of linkage disequilibrium and a large proportion of loci in Nanjenshan and Shouchia that show pairwise locus disequilibrium. We believe continuous genetic drift in the southern populations will increase genetic divergence among populations of C. championii in Taiwan. Significant correlation was found between elevation and expected heterozygosity. We therefore inferred that temperature is the most important ecological factor to influence the genetic diversity of C. championii.     

Genetic diversity and population structure of Yucca filamentosa (Agavaceae)

Using 19 allozyme loci we studied genetic diversity in 18 populations of Yucca filamentosa (Agavaceae) from the southeastern United States. Of the 19 loci surveyed, 17 (89.5%) were polymorphic in at least one of the populations sampled. There was considerable variation among populations in the percentage of polymorphic loci (range = 31.6-84.2%, mean = 67.6%). Similar heterogeneity among populations was observed for mean number of alleles per polymorphic locus (range = 2.0-3.0; mean = 2.48) and mean expected heterozygosity (range = 0.113-0.288; mean = 0.213). On average, 83% of the total genetic diversity was found within populations. Duplications of three allozyme loci were detected in several populations. The life-history characteristics of Y. filamentosa (a long-lived, semiwoody, predominantly outcrossing monocot with a large geographical range) may contribute to the maintenance of such high levels of genetic diversity. These results contradict expectations of the genetic structure of Y. filamentosa based on observations of the dispersal and pollination behavior of its sole pollinator, Tegeticula yuccasella, the yucca moth.     

Patterns and levels of genetic differentiation in North American populations of the Alaskan wheatgrass complex

Levels and distribution of genetic variation were assessed using six allozymes in 27 populations of Alaskan wheatgrass (Elymus alaskanus) from different locations in Canada, USA, Greenland and Russia to obtain information on the genetic structure of these populations. The enzyme systems were ACO, DIA, GPI, MDH, PGM and SKD. Allozyme variation at the species level was high, with 64.3% (Ps) of the loci being polymorphic, an average number of alleles per locus of 1.9 (As), and an average genetic diversity of 0.17 (Hes). Differentiation was found in the populations studied, with the following findings: (1) statistically significant differences were found in allele frequencies among populations for every polymorphic locus (P < 0.001); (2) 63% of the total allozyme variation at polymorphic loci was partitioned among populations (GST = 0.63); (3) relatively low mean genetic distances between the populations were obtained (mean D = 0.029); (4) the genetic structure of Russian populations are clearly distinct from the other populations, the cluster and principal component analyses revealed the same genetic patterns of relationships among populations. This study also indicates that E. alaskanus contains different levels of allozyme variation in its populations. Furthermore, some banding patterns at the loci Aco-1, Aco-2, Gpi-2, Mdh-1, Skd-1, Skd-2 can be used as markers to identify individual populations.     

柚品种的等位酶变异研究

研究了柚的48个品种的等位酶变异,利用等位酶分析技术对柚的酯酶（EST）,6－磷酸葡萄糖异的酶（PGI）,6－磷酸葡萄糖变位酶（PGM）,莽草酸脱氢酶（SKD）,超氧化物歧化酶（SOD）共5个酶系统的10个等位酶基因座进行了分析,除PGI－1,PGI－2两个基因座外,其它8个均为多态性基因座;10个等位酶基因座共观察到的等位基因25个,平均每个基因座的有效等位基因数目为1．55,基因多样度0．2805,柚的品种间具有较为丰富的等位酶标记遗传多样性,但柚类种质资源群体总的遗传多样性水平偏低。柚的较低的有效等位基因数目与基因多样度可能由于人工选择及资源流失造成。     

Enzyme polymorphism in Mansonia uniformis, a mosquito vector of brugian filariasis

Three temporal samples of a wild population of Mansonia uniformis were analysed for genetic variation at six gene-enzyme systems. Adenylate kinase, hexokinase (3 loci) and cathodal malate dehydrogenase were monomorphic. Phosphoglucomutase, glucose phosphate isomerase, isocitrate dehydrogenase and anodal malate dehydrogenase were polymorphic. Each of the polymorphic loci was represented by three alleles. The average heterozygosity or gene diversity was 0.0437.     

Genetic Diversity in Cymophyllus fraserianus (Cyperaceae), a Rare Monotypic Genus

Cymophyllus fraserianus (Fraser's sedge) is a rare perennial evergreen herb found in late-successional forest communities in the Appalachian Mountains of the U.S. Genetic diversity was assessed at 19 allozyme loci for 12 populations sampled from the southern and central portions of the range of this primitive monotypic genus. Species variation was comparable to that found for other narrowly distributed species, although mean population genetic diversity was somewhat higher. Sixteen of the 19 loci (84%) were polymorphic, with a mean of 46% polymorphic within populations. On average, 2.7 alleles were detected per polymorphic locus. Genetic diversity within populations (Hep) averaged 0.113, whereas species' genetic diversity (Hes) was 0.131. Although a twofold range of within-population genetic diversity (Hep = 0.075-0.158) was found, less than 13% of the genetic variation was distributed among populations (G(ST) = 0.129). An indirect estimate of historical levels of gene flow (Nm = 1.69) was consistent with the high mean genetic identity (mean I = 0.96) found between populations. Despite the relictual nature of C. fraserianus, its rarity and fragmented distribution, genetic diversity within the genus is typical of herbs with similar life-history characteristics. No association was found between geographic and genetic distances between populations (r = -0.01; p > 0.9), suggesting that present day populations are relatively isolated.     

Patterns of allozyme diversity in the threatened plant Erigeron parishii (Asteraceae)

Thirty-one occurrences of Erigeron parishii, a narrowly endemic plant threatened by mining, were sampled for allozyme diversity. This taxon held considerable genetic variation at the 14 allozyme loci surveyed. Species (e.g., alleles per locus [A] = 4.3 and proportion of polymorphic loci [P] = 0.64) and population (e.g., A = 2.15 [SD = 0.3] and P = 0.53 [SD = 0.12]) genetic diversity measures were higher than expected for narrowly endemic plant taxa. The proportion of polymorphic loci and numbers of alleles per locus indicated that E. parishii has not experienced severe or long-lasting population bottlenecks. Within-population f indicated low to moderate levels of inbreeding. Populations were only moderately differentiated (theta-p = 0.12), suggesting either that there is substantial gene flow among populations or that populations have not been isolated long enough to detect effects of genetic drift. There was no significant differentiation among populations in different vegetation types nor was there a relationship between genetic distance and geographic distance among sites. Continued fragmentation by mining activities would isolate populations, disrupting gene flow, exacerbating loss of diversity, and increasing inbreeding in the remaining fragments. Protection of large, interconnected populations throughout the range of the taxon is warranted to maintain processes that have sustained the observed diversity.