云南金钱槭形态变异与遗传变异的相关性研究

对我国特有珍稀濒危保护植物云南金钱槭的形态变异水平、遗传变异模式以及两者之间的相关性进行了研究。形态学性状分析结果表明 :各居群形态性状变异系数的平均值从大到小排列为 :文山居群 (WSh)、屏边居群 (PB)、黑龙潭居群 (HL T)、蒙自居群 (MZ) ;文山居群与屏边、黑龙潭、蒙自居群间已产生显著或极显著水平的形态差异 ,而后三者间的差异未达显著水平。RAPD分析检测到 10 3个位点 ,其中多态位点 84个 ,云南金钱槭物种水平的多态位点比率为 81.5 5 % ,与其它珍稀濒危植物相比该种的遗传多样性水平不低。 AMOVA和 N ei基因多样性指数分析显示 ,尽管大部分遗传变异仍存在于居群内 (分别为5 7.86 %、5 7.33% ) ,但居群间的遗传变异已达较高水平 (分别为 4 2 .14 %、4 2 .6 7% )。相关分析结果显示 ,云南金钱槭的形态变异与海拔、土壤有机质等生态因子有着显著或极显著水平的相关性 ,但与遗传变异的相关性未达显著水平 ,说明云南金钱槭的形态变异虽然具有一定的遗传基础 ,但可塑性及环境压力对形态变异的产生作用更大一些。基于形态性状和 RAPD数据的聚类分析则进一步说明 ,云南金钱槭的形态变异受到环境因子的强烈影响而与遗传背景的关系不显著     

云南金钱槭天然居群等位酶遗传多样性研究

云南金钱槭(Dipteronia dyeriana)是我国特有的国家Ⅱ级重点保护珍稀濒危植物,仅分布于云南的文山、屏边和蒙自三县交界地区。采用水平切片淀粉凝胶电泳等位酶分析方法,对采自三个县的天然居群的63个样品进行了遗传多样性检测。8个酶系统的13个等位基因位点分析表明,云南金钱槭的遗传多样性水平低,其平均多态位点百分率P=15.4%,每位点平均等位基因数A=1.2,平均预期杂合度H_e=0.064,各居群都偏离Hardy-Weinberg平衡,杂合子过量,居群间的基因分化系数G_ST=0.099,遗传变异主要发生在居群内(90.1%),居群间分化较小,居群间遗传一致度较高(I=0.985~1),具有一定的基因流(Nm=2.669)。不加权对平均法(UPGMA)聚类可将4个居群分为两支,水头箐居群和马家寨居群亲缘关系最近,聚类后与黑洞居群形成一支再和中槽子居群聚类,这与其地理分布格局大致吻合。据此提出了关于云南金钱槭保育策略的建议。     

中国特有金钱槭属植物物种分化研究

金钱槭属(Dipteronia Oliv.)隶属于槭树科(Aceraceae),是北温带植物区系中古老、残遗的木本属之一,为中国特有属,属下仅包括金钱槭(D.sinensis Oliv.)和云南金钱槭(D.dyerana Henry)2个物种。本实验从形态学、分子遗传学及基因序列分析等方面对金钱槭(D.sinensis Oliv.)和云南金钱槭(D.dyerana Henry)进行对比研究,探讨其物种间分化程度。结果表明:(1)形态学研究显示两物种形态性状存在显著差异,表明其在形态上发生了显著分化;(2)基因组DNA的AFLP标记分析表明,两物种间存在极显著遗传分化(ΦST=0.390 4),在物种水平及群体水平,两物种遗传多样性及遗传变异水平皆有明显差异(金钱槭云南金钱槭);(3)从GenBank数据库中选出2物种已知的叶绿体基因(psbM-trnD、trnD-trnT、trnL、trnL-trnF、rpl16)和核基因ITS(ITS1、5.8SRNA、ITS2)进行序列进化比对分析,t-test检验表明,2物种叶绿体基因序列间有极显著差异。研究结果表明:金钱槭和云南金钱槭两物种间产生了极显著的分化,推测可能由于长期的地理隔离所致。     

云南金钱槭 等位酶 遗传多样性 生境片段化 保育策略

云南金钱槭（Dipteronia dyeriana）是我国特有的国家Ⅱ级重点保护珍稀濒危植物,仅分布于云南的文山、屏边和蒙自三县交界地区。采用水平切片淀粉凝胶电泳等位酶分析方法,对采自三个县的天然居群的63个样品进行了遗传多样性检测。8个酶系统的13个等位基因位点分析表明,云南金钱槭的遗传多样性水平低,其平均多态位点百分率P=15.4%,每位点平均等位基因数A=1.2,平均预期杂合度He=0.064,各居群都偏离Hardy-Weinberg平衡,杂合子过量,居群间的基因分化系数GST=0.099,遗传变异主要发生在居群内（90.1%）,居群间分化较小,居群间遗传一致度较高（I=0.985～1）,具有一定的基因流（Nm=2.669）。不加权对平均法（UPGMA）聚类可将4个居群分为两支,水头箐居群和马家寨居群亲缘关系最近,聚类后与黑洞居群形成一支再和中槽子居群聚类,这与其地理分布格局大致吻合。据此提出了关于云南金钱槭保育策略的建议。     

金钱槭属植物的遗传多样性与保护策略

金钱槭属包括2个种:金钱槭(Dipteronia sinensis Oliv.)和云南金钱槭(Dipteronia dyeriana Henry),均为珍稀濒危植物,主要分布于中国的中部和南部。该研究应用18种随机扩增多态性DNA(RAPD)引物、8对扩增片段长度多态性(AFLP)引物和10种多态性叶绿体微卫星基因(cpSSR)引物,获得金钱槭和云南金钱槭的遗传多样性与种群结构的相关信息。结果表明,在物种水平上,由RAPD(金钱槭:H=0.386 4,Hsh=0.556 3;云南金钱槭:H=0.304 7,Hsh=0.445 0)和AFLP(金钱槭:H=0.331 9,Hsh=0.488 0;云南金钱槭:H=0.304 7,Hsh=0.4450)2个核标记的遗传多样性具有可比性,而cpSSR标记(金钱槭He=0.603 2,云南金钱槭He=0.671 1)的遗传多样性高于AFLP和RAPD标记的遗传多样性,遗传分化系数(GST)也显示出相似的结果,由此揭示2个种均拥有相对较高的遗传多样性和遗传分化。研究认为,人类活动破坏植物栖息地是造成2种物种处于濒危状态的主要原因;从长远来看,保护2种金钱槭属植物的主要策略是保护植物栖息地,同时应加强扩大种群规模以维持现有遗传变异水平。     

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。     

云南南部不同种源地小桐子遗传多样性的ISSR分析

应用ISSR分子标记方法对采自云南的8个居群的小桐子(Jatropha curcas)共158个个体进行遗传多样性分析。8个ISSR引物共扩增到了67个位点,其中61个是多态性位点。分析结果表明:(1)云南小桐子的遗传多样性水平很高。在物种水平上,平均每个位点的多态位点百分率PPB=91.04%,有效等位基因数Ne=1.5244,Nei′s基因多样性指数He=0.3070,Shannon多样性信息指数Ho=0.4618;在居群水平上,PPB=55.04%,Ne=1.3826,He=0.2171,Shannon多样性信息指数Ho=0.3178。(2)居群间的遗传分化低于居群内的遗传分化。基于Nei′s遗传多样性分析得出的居群间遗传多样性分化系数Gst=0.2944。AMOVA分析显示:云南小桐子的遗传变异主要存在于居群内,占总变异的63.50%,居群间的遗传变异占36.50%。(3)居群间的地理距离及遗传一致度并不存在相关性。鉴于以上指标,我们推测云南小桐子可能来自不同的地区。     

准噶尔无叶豆片断化居群的遗传变异及克隆多样性

准噶尔无叶豆(Eremosparton songoricum)是豆科无叶豆属小半灌木, 既能开花结实进行有性繁殖, 又可以靠根茎进行无性克隆繁殖, 为国家三级保护植物, 在中国仅片断化分布于新疆古尔班通古特沙漠局部区域。本文采用ISSR分子标记对采自古尔班通古特沙漠腹地及边缘的7个准噶尔无叶豆自然居群共148个个体进行了遗传变异和克隆多样性分析。8个引物共扩增出84个位点, 其中77个为多态性位点, 物种水平上的多态位点百分比PPB为91.67％, Nei's基因多样性指数I为0.3192, Shannon信息指数H为0.3540; 居群水平上的多态位点百分比PPB为58.45％, Nei's基因多样性指数I为0.2248, Shannon信息指数H为0.3270。居群间的遗传分化系数G_ST为0.2978。AMOVA分析表明, 有31.88%的遗传变异存在于居群间, 表明居群间存在显著的遗传分化。克隆多样性分析表明, 居群水平上, 居群G的Simpson多样性指数和均匀度指数最高, 分别为0.9400和0.9885; 居群E最低, 分别为0.8457和0.9021。物种水平上, Simpson多样性指数为0.9858, 均匀度指数为0.9673。本研究结果表明, 和其他荒漠植物相比, 准噶尔无叶豆表现出较高的遗传变异水平和克隆多样性, 这主要与该物种兼性生殖的繁育方式及多克隆起源有关; 而居群间产生了显著的遗传分化则主要由于人为干扰引起的生境片断化和居群减小而导致的基因交流障碍所致。遗传变异水平和遗传结构的研究将为分析准噶尔无叶豆致濒原因及进化潜力提供重要资料, 对该物种保护具有指导意义。     

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.     

云南金钱槭果实、种子形态分化研究

对5个云南金钱槭(Dipteronia dyeriana Henry)天然居群的果实、种子形态进行研究.测量了果实和种子的长度、宽度、重量等12个表型性状指标.数据统计结果显示5个云南金钱槭居群按果实、种子大小进行排序为蒙自(MZ)>屏边(PB)>文山1(WSh1)>文山3(WSh3)>文山2(WSh2).表型性状在居群内和居群间均存在着一定程度的变异,其中居群间变异系数的平均值从0.064(果实整体形态,PL/PB)到0.197(种子重量,SW),相比之下果实整体形态最为稳定.居群间形态总体差异显著性配对t检验结果说明,多数居群间已产生较明显的形态分化.Ward聚类和相关分析结果都表明表型性状与生态因子、海拔高度之间存在着一定程度的相关性,多种生态因子的共同作用是导致果实、种子形态特征产生差异的主要原因.     

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.     

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. __________ Translated from Acta Phytoecologica Sinica, 2005, 29(5): 785–792 [译自: 植物生态学报, 2005, 29(5): 785–792]     

Genetic Diversity of Jatropha curcas (Euphorbiaceae) Collected from Southern Yunnan,Detected by Inter-simple Sequence Repeat ( ISSR)

The genetic diversity of 158 individuals from eight semi-wild populations from Yunnan Province was estimated using ISSR method (8 primers). The results revealed an extraordinarily high level of genetic diversity ( at species level,percentage of polymorphic loci PPB = 91.04% , effective number of alleles N_e = 1.5244 , Nei′s (1973 ) gene diversity H_e= 0.3070, and Shannon′s information index H_o = 0 . 4618 ; at population level, PPB = 55. 04% , N_e = 1.3826, Nei′s (1973) gene diversity H_e = 0.2171, and Shannon′s information index H_o = 0.3178). The level of genetic differentiation between populations is lower than that among populations . The low level of genetic differentiation among populations was detected, based on Nei′s genetic diversity analysis (29.44%), and AMOVA (36.50%). There is no associations between geographical distance and genetic identity.We suggest that Jatropha curcas of Yunnan Province might not be introduced from the same place.     

云南南部不同种源地小桐子遗传多样性的ISSR 分析

应用ISSR 分子标记方法对采自云南的8 个居群的小桐子( Jatropha curcas) 共158 个个体进行遗传多样性分析。8 个ISSR 引物共扩增到了67 个位点, 其中61 个是多态性位点。分析结果表明: (1) 云南小桐子的遗传多样性水平很高。在物种水平上, 平均每个位点的多态位点百分率PPB = 91.04% , 有效等位基因数N_e = 1.5244, Nei′s 基因多样性指数H_e= 0.3070, Shannon 多样性信息指数H_o = 0.4618; 在居群水平上, PPB = 55.04%, N_e = 1.3826, H_e = 0.2171, Shannon 多样性信息指数H_o = 0.3178。(2) 居群间的遗传分化低于居群内的遗传分化。基于Nei''s 遗传多样性分析得出的居群间遗传多样性分化系数G_st = 0.2944。AMOVA分析显示: 云南小桐子的遗传变异主要存在于居群内, 占总变异的63.50%, 居群间的遗传变异占36.50%。(3) 居群间的地理距离及遗传一致度并不存在相关性。鉴于以上指标, 我们推测云南小桐子可能来自不同的地区。     

秋海棠属植物种质亲缘关系的ISSR分析

应用ISSR标记方法对秋海棠属(Begonia L.)植物33个种的种质亲缘关系进行分析,从70条引物中筛选出13个多态性引物,并通过POPgen 32软件以UPGMA法分析其亲缘关系。结果表明:33份秋海棠属植物的遗传距离在0.188 9～0.932 8之间,多态性百分率达到97.54%,Shannon信息指数为0.551 9,Nei基因多样性指数为0.376 0。秋海棠属植物具有丰富的遗传变异,UPGMA聚类分析结果与现有属下分类学表现较为一致,且生态地理条件相似的种群优先聚集。     

山东省长岛县南部四岛藜（Chenopodium album L.）天然种群遗传多样性的ISSR分析

采用ISSR分子标记对分布于山东省长岛县南部的北长山、南长山、大黑山和小黑山4个岛屿上的藜天然种群共81个个体进行了遗传多样性及遗传结构的研究。13个引物共检测到157个可重复的位点。遗传多样性研究结果表明：种内的多态位点比率为66.24％,具有较高的遗传多样性（Shannon（I）指数在物种水平上为0.332 0）;种群间有一定的遗传分化,根据G_st值,种群间的遗传多样性占总群体的9.27％。遗传距离分析表明,XHS种群和NCS种群的遗传一致度最高,与地理距离无相关性。     

Genetic diversity analysis of Hepatacodium miconioides at different altitude in Tiantal Mountain,Zhejiang Province,China and its relationship with environmental factors

The genetic diversity within and among populations of Hepatacodium miconioides collected at three different altitudes in Tiantai Mountain,Zhejiang Province and its relationships to environmental factors were analyzed by random amplified polymorphic DNA(RAPD)technique.Amplification using 12 random primers of 60 plants and 122 repetitive loci were produced.The percentage of polymorphic loci of three populations ranged from 18.85% to 23.77% with an average of 21.86%,indicating the relatively low genetic diversity of H.miconioides.The average Shannon index of phenotypic diversity(0.1329)and Nei index(0.0925)within populations were relatively low.A distinct genetic differentiation existed among populations Of H miconioides in spite of the relatively small geographical distribufion.The average genetic diversity within populations of H.miconioides accounted for 33.58% of the total genetic diversity while the genetic diversity among populations accounted for 66.42% as estimated by the Shannon index of phenotypic diversity,The genetic differentiation among populations of H.miconioides was 0.6546,as estimated by Nei index.The gene flow estimated from Gsr was only 0.2656 and it indicated that gene flow among populations of H.miconioides was relatively low.The mean value of the genetic identity among populations of H.miconioides was 0.7126 and the average of genetic distance of H.miconioides was 0.3412.The genetic identity between populations at the elevation of 990m and at the elevation of 780 m was the highest.The genetic identity between population at the elevation 500 m and other two populations was relatively low.The correlation analysis showed that the genetic diversity within populations was significantly related with the soil total nitrogen.     

7种川产淫羊藿属植物遗传多样性及亲缘关系的ISSR分析

利用ISSR分子标记对7种12居群的川产淫羊藿属植物进行遗传多样性及亲缘关系分析。结果发现23个ISSR随机引物共扩增194条清晰条带,其中169条具多态性,平均多态性位点比率为87.11%,有效等位基因数Ne=1.534 2,Nei基因多样性指数H=0.314 4,Shannon多样性指数I=0.469 7,表明物种间遗传多样性丰富。7种川产淫羊藿属植物间的遗传相似系数为0.653 2～0.748 9,聚类分析和主成分分析直观的显示出12份供试材料间的亲缘关系,并将其分为两类。表明ISSR分子标记技术可以用于川产淫羊藿属植物的遗传多样性和亲缘关系分析。     

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。