珍稀濒危蕨类植物东方水韭的遗传多样性

采用ISSR分子标记对珍稀濒危蕨类植物东方水韭和中华水韭4个自然居群共72个样品进行了DNA多态性分析。从65个随机引物中筛选出11个有效引物,共产生98条DNA片段,其中68条为多态性条带,多态位点百分率(PPL)为66.33%。东方水韭两个居群的多态位点百分比率(PPL)较低(8.16%和7.14%)。中华水韭两个居群的PPL也较低(9.18%和23.47%)。中华水韭物种水平上的PPL(51.02%)高于东方水韭(9.18%)。AMO-VA分析结果表明,东方水韭遗传变异中主要存在于居群内(94.74%),而中华水韭绝大部分的遗传变异却存在于居群间(86.71%)。东方水韭和中华水韭个体间UPGMA聚类结果表明:同一物种的个体完全能聚在一起,中华水韭的两个居群也能明显分开,而东方水韭两居群的个体并不能完全聚在一起。探讨了可能造成上述居群遗传结构模式的主要因素,同时提出今后工作中需重点解决的问题。     

利用ISSR标记对新疆白梭梭居群的遗传多样性分析

利用ISSR分子标记对新疆白梭梭4个居群,105个个体进行了遗传多样性的比较分析。在供试材料中,11个引物共扩增出171个多态位点,多态位点百分率为84．85％,4个居群的多态位点百分率差异在33．92％．40．35％之间。Shannon多样性指数（I）为0．3518,物种水平的Nei基因多样度（h）为0．3482。遗传变异分析表明,物种水平的居群间遗传分化系数Gst为0．6238,居群间的基因流Nm为0．3016。遗传分析表明吐鲁番居群和甘家湖居群的遗传距离最近。     

兰花蕉的遗传多样性研究

采用简单重复序列区间(ISSR,Inter-Simple Sequence Repeat)分子标记技术,对采自广东省的濒危植物兰花蕉(Orchidantha chinensisT.L.Wu)的7个居群137个个体进行遗传变异分析。用10个ISSR引物共扩增出清晰谱带101条,其中58条具有多态性,总多态位点百分率为57.43%。居群水平相对较低,多态位点百分率在6.93%-35.64%之间,平均为18.24%。经POPGENE1.31数据处理,结果表明:在物种水平上Nei基因多样性为0.1254±0.1686;Shannon信息指数为0.2000±0.2429;Nei基因分化系数为0.5481,表明54.81%的遗传变异分布在居群间,45.19%的遗传变异分布在居群内。物种居群间的遗传一致度在0.8855-0.9511之间。我们认为红花潭是其最适合生境,建议在此建立自然保护区;鉴于兰花蕉居群间出现了一定程度的分化,为最大限度地保护兰花蕉的遗传多样性,建议在自然居群间进行相互移栽,以提高群体间的基因交流。     

西双版纳地区流苏石斛遗传多样性的ISSR分析

采用ISSR分子标记技术,对西双版纳分布的兰科濒危植物流苏石斛（Dendrobium fimbriatum）5个居群共114个个体的遗传多样性进行了研究。从100条引物中筛选出了12条用于扩增,共检测到117个位点,其中105个为多态位点。分析结果表明,流苏石斛居群水平遗传多样性较低。在物种水平上,流苏石斛多态位点百分率PPB为89．74％,Nei’s基因多样性指数日为0．3227,Shannon’s多样性信息指数见。为0．4779;在居群水平上,各个居群的多态位点百分率PPB差异较大（6．84％～39．32％）,平均值为23．93％,Nei’s基因多样性指数H为0．0871,各个居群的Shannon’s多样性信息指数见平均为0．1290。AMOVA分析的结果显示,流苏石斛的遗传变异大多数存在于居群间,占总遗传变异的74．79％。基于Nei’s遗传多样性分析得出的居群间遗传分化系数Gst=0．7443。各居群间的Nei’s遗传一致度（I）范围为0．5882～0．8331。Mantel检测发现,居群间的遗传距离和地理距离之间无显著的正相关关系（r=0．2419,P=0．2416）。鉴于流苏石斛的遗传多样性现状和居群遗传结构,我们建议对流苏石斛居群所有个体实施及时的就地保护,同时建立迁地保护居群,促进基因交流。     

古尔班通古特沙漠荒漠肉苁蓉遗传多样性分析

为了全面了解古尔班通古特沙漠荒漠肉苁蓉居群分布的遗传多样性特点,本研究通过ISSR分子标记技术,利用Nei和Shannon等多样性指数对古尔班通古特沙漠中5个居群166个个体的荒漠肉苁蓉遗传多样性、荒漠肉苁蓉种群和种内的遗传多样性进行分析。在供试材料中,8个引物共扩增出144个多态位点,多态位点百分率达100%,5个居群的多态位点百分率差异在46.53%～77.78%之间。在物种水平上,Nei基因多样度(h)为0.260 4,Shannon多样性指数(I)是0.411 0。遗传变异分析表明,物种水平的居群间遗传分化系数Gst为0.222 2,居群间的基因流Nm为1.750 7。研究显示古尔班通古特沙漠中荒漠肉苁蓉多态位点比例高,各居群基因交流较多,不同居群间遗传变异并不明显,这些对肉从蓉资源有效地保护和利用具有重要意义。     

红泡刺藤居群的遗传多样性研究

利用ISSR分子标记对红泡刺藤的12个居群共242个个体进行了遗传多样性分析.结果表明:(1)16个ISSR引物共扩增到257个位点,其中236个是多态性位点,占91.83％.(2)红泡刺藤居群具有较高水平的遗传多样性,在物种水平上,平均每个位点的多态位点百分率为97.50％,有效等位基因数为1.267,Nei's遗传多样性为0.177,Shannon's多态信息指数为0.296;居群水平上多态位点百分率为51.43％,有效等位基因数为1.205,Nei's 遗传多样性为0.127,Shannon's多态信息指数为0.202.(3)居群间基因分化系数为0.2815,AMOVA分析居群间遗传变异占总量的34.47％,二者结果相近,说明红泡刺藤居群间存在一定程度的遗传分化.居群内的遗传变异为65.53％,基因流为1.2762.(4) Mantel检测显示居群间的遗传距离与地理距离不存在相关性.UPGMA聚类分析和二维主成分分析结果一致,红泡刺藤居群可分为2个居群组,即金沙江居群和维西居群为一个类群,其他居群为另外一大类,表明生态地理条件相似的居群优先集中.     

刺槐不同居群遗传多样性的ISSR分析

利用ISSR标记对全国10个刺槐居群子代100个个体的遗传多样性进行了比较分析,从65个随机引物中筛选出10个多态性引物进行扩增,共检测到91个位点,多态位点数(AP)为85,多态位点百分率(P)为93.41%.刺槐在种级水平的遗传多样性参数略高于居群水平,多态位点百分率(P)分别为95.60%、69.01%,Shannon′s信息指数(I)分别为0.6145、0.3733,Nei′s基因多样性指数(H)分别为0.4337、0.2514.居群间的遗传分化指数Gst、Nei′s基因多样性指数和Shannon′s信息指数统计结果,均显示出中国刺槐居群内遗传多样性大于居群间遗传多样性.利用PopGen32软件对10个居群进行聚类分析可知,10个刺槐群体可分为三大类,亲缘关系和地理分布呈一定的相关性,但没有形成明显的地理变异模式.     

华中特有珍稀植物裸芸香的AFLP遗传多样性分析

采用选择性扩增片段多态性(AFLP)方法对华中特有单种属植物裸芸香(Psilopeganum sinense)的8个自然居群的遗传多样性进行了检测与分析。结果表明:裸芸香的遗传多样性较低,且居群内遗传多样性显著低于物种水平遗传多样性。筛选出的5对引物共得到180个位点,76个为多态位点,多态位点百分率为42.2%,8个居群多态位点百分率为:3.3%～16.7%,居群平均多态位点百分率为9.4%;8个居群Nei多样性指数为0.01987～0.06987,Shannon’s多样性指数为0.0197～0.0816。居群间分化系数Gst=0.5069,居群间基因流为0.2432,不足以维持居群间的基因交流及现有的遗传结构。AMOVA分析表明总遗传变异的13.17%存在于4个地理区域之间,50.45%存在于地理区域内的居群间,36.38%的遗传变异存在于居群内个体间。NTSYS分析表明遗传距离与地理距离不存在相关关系。UPGMA聚类结果表明长江南北两岸的居群并没有产生明显分化。最后,分析了裸芸香的濒危原因并提出了有效的保育措施。     

珍稀濒危植物天目木兰（Magnolia amoena）遗传多样性的RAPD分析

运用随机扩增多态DNA(RAPD)技术,对天目木兰(Magnolia amoena)居群的遗传多样性进行了研究．从40个10-mer随机引物中筛选出14个能得到清晰、稳定扩增带的引物进行扩增,14个引物共检测了94个位点,其中多态性位点为23,占24．4％,计算了12个居群之间的遗传相似度和遗传距离,并运用UPGMA法进行了聚类分析,结果显示相同严地个体间(居群内)的遗传距离较小,遗传多样性水平很低;不同产地个体间(居群间)遗传距离较大,遗传多样性水平较前者高,即天目木兰个体间遗传多样性水平与它的地理分布有关,天目木兰总体较低的遗传多样性是导致它濒危的原因之一。     

云南穗花杉的遗传多样性研究

采用随机扩增多态DNA(RAPD)方法对云南穗花杉(Amentotaxus yunnanensis)4个居群和台湾穗花杉似(Amentotaxus for9rmosana)1个居群共104个个体进行了遗传多样性分析。9个随机引物共扩增出清晰谱带143条。云南穗花杉在物种水平上遗传多样性较高(多态位点百分率P为79．0％,基因多样性指数He为0．2718),但云南穗花杉和台湾穗花杉居群内遗传多样性均较低(P为18．0％、6．9％;He为0．0688、0．0198)。云南穗花杉居群间遗传分化强烈(AMOVA,GST和Shannon多样性指数分别为0．7611,0．7503和0．7526)。据推测,第四纪冰川引起的瓶颈效应,小规模居群引起的遗传漂变及幼苗成活率低等因素都加剧了居群间的遗传分化。建议对所研究的云南穗花杉全部居群予以保护,特别是对云南西畴和贵州兴义市七舍两个具有相对较高遗传多样性的居群应该优先建立就地保护点,以达到最大限度保存云南穗花杉遗传多样性的目的。     

大别山山核桃天然群体遗传结构的初步分析

利用RAPD分子标记技术检测了大别山山核桃3个天然居群的遗传多样性和遗传结构。20条10 bp随机引物共检测到238个扩增位点,其中多态性位点162个,多态位点百分率(PPB)为68.1%。居群水平Shannon’s多态性信息指数(I)介于0.2651~0.2801之间;居群水平Ne i’s基因多样性指数(H)介于0.1789~0.1890之间。遗传变异计算显示大别山山核桃居群间基因分化系数(Gst)为0.4063,分子方差分析(AMOVA)表明居群间基因分化水平为0.4177,居群间基因流(Nm)为0.7306,说明大别山山核桃大部分变异存在于居群内,居群间基因交流相对较少。这一结果符合大别山山核桃风媒、异交的繁育系统特点,但其居群间基因分化程度明显高于异交植物的平均水平(Gst=0.1930)。地理隔离、居群内近交及居群间基因流受阻可能是形成目前大别山山核桃天然群体遗传结构的主要因素。     

中国特有植物卧龙沙棘自然群体的RAPD分析

应用12个随机引物对卧龙沙棘全部2居群共28个个体进行了RAPD分析。结果表明,卧龙沙棘具有较丰富的遗传多样性,多态位点百分率为78.05%,Nei’s基因多样性h=0.2553,Shannon’s多态性信息指数Ι=0.3841。分布范围狭窄的卧龙沙棘在亚种水平的遗传多样性明显高于分布较广的中国沙棘。在居群水平上,卧龙沙棘同样具有很高的遗传多样性,平均多态位点百分率为63.42%,Nei’s基因多样性h=0.2193,Shannon’s多态性信息指数I=0.3287。卧龙沙棘的基因分化系数Gst=0.1425,表明遗传变异主要存在于居群内。AMOVA的结果也表明,在全部的遗传变异中,19.18%的遗传变异存在于居群之间,与Gst值基本一致。这一结果符合卧龙沙棘风媒、异交的繁育系统特点。     

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。     

Genetic variability and differentiation of Caragana microphylla populations as revealed by RAPD markers

Genetic variability in random amplified polymorphic DNA (RAPD) was studied in 90 individuals of Caragana microphylla, an outcrossing perennial shrub species, from five natural populations sampled in Inner Mongolia steppe of China on a small scale. Nineteen selected primers were used to amplify DNA samples, and totally 225 bands were detected. The percentage of polymorphic bands within populations ranged form 58.22% to 63.56%, with an average of 60% at the population level and 71.11% at the species level, indicating relatively high genetic variations in C. microphylla species. Shannon's information index (I) and Nei's gene diversity (h) showed the similar trend with each other. According to the analysis of Nei's gene diversity, the percentage of genetic variation among populations was 7.13%, indicating a low level of genetic differentiation among populations. There existed a strong gene flow (Nm = 3.26) among populations. Although AMOVA analysis also revealed most variation was within populations (phi(ST) = 4.1%), a significant proportion was observed among populations (P<0.001) in the present study, suggesting genetic differentiation occurred among populations at a certain extent. Based on Mantel's tests and the results of previous studies, the genetic structure pattern of C. microphylla accorded with the isolation-by-distance model on a very large scale, however, on a small scale, the significant genetic differentiation among populations might be enhanced by the micro-environmental divergence among the sampling sites, rather than by geographic factors. Analysis of the genetic variations of C. microphylla populations provided useful information for the adaptive strategy of Caragana species.     

内蒙古高原小叶锦鸡儿(Caragana microphylla)、中间锦鸡儿(C. davazamcii)和柠条锦鸡儿(C. korshinskii)的遗传多样性及遗传关系

采用RAPD技术对10个具较大地理跨度的小叶锦鸡儿、中间锦鸡儿和柠条锦鸡儿种群的遗传多样性和遗传关系进行了研究.共检测到678个位点,多态条带比率(PPB)为100%;特有位点41个,占6.05%.总体上3种锦鸡儿的遗传多样性表现出自东向西递减的趋势,分析表明其与生长地点年均气温呈显著负相关.AMOVA表明:3种锦鸡儿种间变异只占总体变异的6.08%,且显著性检验表明这种变异不显著;种内种群间的变异占总变异的11.90%;总变异的主要部分来自种群内部(82.02%). 3种锦鸡儿各种群总体分析结果表明:种群内变异比率Hpop/Hsp为0.8013,基因分化系数Gst为0.1603,种群每代迁移数Nm为2.6192,显示种群间存在一定强度的基因流,3种锦鸡儿间表现为异交性.3种锦鸡儿多样性高低及种群聚类分布格局都表现出一定的地理连续性.     

Genetic diversity and genetic relationship of Caragana microphylla, Caragana davazamcii and Caragana korshinskii on the Inner Mongolia Plateau, China

C. microphylla, C. davazamcii and C. korshinskii exhibit a geographical replacement series from east to west on the Inner Mongolia Plateau. Currently, there is still a debate about the taxonomic and genetic relationship among these 3 species. We studied the genetic diversity and genetic relationship among these 3 species by analyzing DNA samples of individual plants from within 10 populations with random amplified polymorphic DNA (RAPD) markers. We identified 678 RAPD loci in total, of which all were polymorphic (PPB = 100%). There were 41 unique loci (6.05%). In general, a trend presented that the genetic diversity of these species decreased from east to west. Further, the genetic diversity was significantly negatively correlated with the local annual mean temperature. Analysis of molecular variation (AMOVA) showed that the genetic variation among these 3 species was only 6.08% of the total genetic variation. Between the species, the genetic variation was insignificant (P = 0.9961). The proportion of genetic variation among populations within each species was 11.90% (P < 0.001) of the total genetic variation, and the total genetic variation mainly existed within the populations (82.02%). Estimated with Shannon's index, genetic differentiation within the populations (Hpop/Hsp) was 0.8013, the coefficient of gene differentiation (Gst) was 0.1603, and the gene flow index (Nm) was 2.6192. This, thus, indicates that there is relatively high gene flow among these populations, and that these 3 species are crossbreeding. The genetic diversity level and the population distribution pattern showed geographic continuity to some extent.     

有性生殖对栗疫病菌群体结构的影响

采用RAPD方法对来源于栗疫病菌8个不同子囊壳的子囊孢子后代和无性生殖的对照群体各23个菌株进行了群体结构的比较。从RAPD随机引物中筛选出扩增多态性丰富的4条引物,共扩增出条带73条,多态性检测率为100%。研究结果表明,在8个子囊壳和无性生殖群体中的基因多样性,64.27%由群体内部引起,只有35.73%的多样性由群体之间的基因差异引起。各子囊壳群体间存在的基因流动很小(Nm=0.8994)。有性群体和无性群体之间的遗传距离为0.1389,基因流动值为3.4212,说明子囊壳群体和无性生殖群体之间存在一定的系统关系。分析表明栗疫病菌子囊孢子后代在自然界的传播对自然界的病菌的多样性起重要的作用。     

濒危植物长叶榧群体遗传多样性的RAPD分析

借助随机扩增多态DNA方法,分析了中国特有的濒危植物长叶榧的遗传多样性和遗传分化．结果表明：12个随机引物在9个长叶榧自然群体180个样品中可检测到180个可重复位点,其中多态位点119个．长叶榧物种水平的遗传多样性较高,多态位点百分率（P）为66．11％,Shannon信息指数（,）为0．3087,Nei指数（h）为0．2015;而群体水平的遗传多样性较低,P、I和h分别平均为23．76％、0．1221和0．0813．AMOVA分子变异显示,42．57％变异来源于群体内,57．43％变异来源于群体间,群体间的遗传分化系数（Gst）为0．5965,群体间的遗传分化程度高．长叶榧群体间的基因流很低,为0．3382．瓶颈效应、群体隔离和群体间基因流低等因素都加剧了长叶榧群体间的遗传分化．9个长叶榧群体间的平均遗传距离为0．1630．通过UPGMA进行聚类,可将9个长叶榧群体分为浙江和福建两大类群．建议在迁地保护时应尽量避免在群体之间实施种质迁移．     

A comparison of the genetic diversity in Dipteronia sinensis Oliv. and Dipteronia dyeriana Henry

Dipteronia is an endemic genus to China and includes only two species, Dipteronia sinensis and D.dyeriana.Based on random amplified polymorphic DNA (RAPD) markers,a comparative study of the genetic diversity and genetic structure of Dipteronia was performed.In total,128 and 103 loci were detected in 17 D.sinensis populations and 4 D.dyeriana populations,respectively,using 18 random primers.These results showed that the proportions of polymorphic loci for the two species were 92.97% and 81.55%,respectively,indicating that the genetic diversity of D.sinensis was higher than that of D.dyeriana.Analysis,based on similarity coefficients,Shannon diversity index and Nei gene diversity index,also confirmed this result.AMOVA analysis demonstrated that the genetic variation of D.sinensis within and among populations accounted for 56.89% and 43.11% of the total variation,respectively,and that of D.dyeriana was 57.86% and 42.14%,respectively.The Shannon diversity index and Nei gene diversity index showed similar results.The abovementioned characteristics indicated that the genetic diversity levels of these two species were extremely similar and that the interpopulational genetic differentiation within both species was relatively high.Analysis of the genetic distance among populations also supported this conclusion.Low levels of interpopulational gene flow within both species were believed to be among the leading causes for the above-mentioned phenomenon.The correlation analysis between genetic and geographical distances showed the existence of a remarkably significant correlation between the genetic distance and the longitudinal difference among populations of D.sinensis (p＜0.01),while no significant correlation was found between genetic and geographical distances among populations of D.dyeriana.This indicated that genetic distance was correlated with geographical distances on a large scale rather than on a small scale.This result may be related to differences in the selection pressure on species by their habitats with different distribution ranges.We suggest that in situ conservation efforts should focus on establishing more sites to protect the natural populations and their habitats.Ex situ conservation efforts should focus on enhancing the exchange of seeds and seedlings among populations to facilitate gene exchange and recombination,and to help conserve genetic diversity.     

内蒙古典型草原羊草种群遗传分化的RAPD分析

运用 RAPD技术对内蒙古典型草原不同生境 8个羊草种群进行分析。采用 2 4个随机引物 (10 nt)在 8个种群中共检测到2 2 4个扩增片断 ,其中多态性片断 173个 ,总的多态位点百分率达 77.2 % ,特异性片断 2 2个 ,占 9.82 % ,平均每个引物扩增的DNA带数为 9.3 3条。利用 Nei指数和 Shannon指数估算了 8个种群的遗传多样性 ,并计算种群相似系数和遗传距离 ,运用UPGMA法进行聚类分析。结果表明 :羊草大部分的遗传变异存在于种群内 ,只有少部分的遗传变异存在于种群间 ,Nei指数和Shannon指数计算结果分别为 85.4%和 66.8% ;羊草不同种群的遗传多样性存在差异 ;8个羊草种群平均遗传距离为 0 .2 3 16,变异范围为 0 .1587～ 0 .2 70 0 ,说明 8个羊草种群间的遗传变异不大 ,即 :在较小地理范围内羊草的遗传分化程度较小 ;8个种群可聚为 3个类群 ,聚类结果显示生境相似的种群能够聚在一起 ,而地理距离最近的种群不一定归为一类 ,说明小范围内羊草种群间的遗传分化与地理距离不存在相关性 ,而与其生境间的相似度相关。影响遗传相似性的不是单一因子而是各种因子的综合作用 ,较小地理范围内羊草种群间的遗传分化主要是由环境的异质性所引起的