金钱槭和云南金钱槭遗传多样性比较研究

金钱槭属(Dipteronia)是我国特有少种属,属下仅金钱槭(D. sinensis)和云南金钱槭(D. dyeriana)两种。该文用RAPD标记揭示了金钱槭的遗传多样性和遗传结构,并与云南金钱槭的RAPD研究结果进行了比较。同时,对两物种遗传距离与地理距离的相关性进行了分析,结果有助于阐释该属植物遗传变异的产生机制。研究显示,18条随机引物在17个金钱槭居群(226个个体)中检测到128个扩增位点,物种水平的多态位点比率为92.97%,在4个云南金钱槭居群(45个个体)中则检测到103个扩增位点,物种水平的多态位点比率为81.55%,金钱槭的多态位点比率高于云南金钱槭。相似性系数值、Shannon多样性指数和Nei基因多样性指数分析反映了与多态位点比率相一致的结果。AMOVA(Analysis of molecular variance)分析结果显示,金钱槭居群内、居群间的遗传变异分别占总变异量的56.89%和43.11%。云南金钱槭居群内、居群间的遗传变异分别占总变异量的57.86 %和42.14%。Shannon多样性指数、Nei基因多样性指数的分析结果与AMOVA分析结果趋势相同。上述特征值揭示,金钱槭和云南金钱槭居群间的遗传分化均已达到较高水平,推测居群间低水平的基因流可能是导致上述现象产生的原因之一。遗传距离与地理距离的相关分析结果显示,金钱槭居群间的遗传距离与经度差异存在极显著水平的相关性(p<0.01),云南金钱槭居群间的遗传距离与地理隔离则无显著相关关系。说明在大尺度上遗传距离与地理距离相关而在小范围内则无上述关系,该结果可能与位于不同分布区内的物种所承受的生境选择压力不同有关。建议在对该属植物进行就地保护时,应设立多个保护点,保护自然居群及其周围生境;在迁地保护时,应通过加大居群间种子和幼苗的交换,人为创造基因交流和重组的条件,保存该属植物的遗传多样性。     

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     

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]     

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。     

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。     

稀有植物云南金钱槭生长群落特征与保护对策

稀有植物云南金钱槭（Dipteronia dyeriana）分布在云南文山老君山,该山在地理位置上处于亚热带南部。但云南金钱槭仅生长在山体的中上部,海拔1800～2200m,生长地因海拔高,具有温凉、湿润的气候特点。云南金钱槭喜光,不适应森林内部环境。老君山云南金钱槭的大多数个体是以单优灌木群落形式存在,少量生长在灌丛中和森林边缘。单优灌木群落是干扰的产物,也由于干扰的延续而存在。分布区森林乔小被砍伐不会导致云南金钱槭消失,原生森林植被完全恢复反而会减少个体数量。对云南金钱槭生存最大的威胁是农地扩大。要维持较高个体数量,适当的干扰是必需的。     

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.     

省沽油群体遗传多样性的AFLP分析

利用4对AFLP引物对来自安徽石台、湖北大悟和河南桐柏的3个省沽油群体进行遗传多样性分析,共扩增出489条带,其中多态性条带460条,多态性百分比为93.99%。不同群体Nei’s基因多样性指数(H)和Shannon’s信息指数(I)变化范围分别为0.1920～0.2046和0.2937～0.3151,其中湖北大悟群体遗传多样性最高。物种水平和种群水平的H分别为0.2190和0.1964,群体内变异占总变异的89.68%,表明省沽油遗传变异主要存在于各群体内部。3个群体的平均遗传距离为0.0292,UPGMA聚类分析结果说明省沽油各群体间亲缘关系较近并和地域具有相关性。建议省沽油的遗传资源保护应以种内遗传多样性的保护为主。     

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.     

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H 为0 . 3227 , Shannon′s 多样性信息指数Hsp 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Ho 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。     

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。     

Genetic diversity analysis of Hepatacodium miconioides at different altitude in Tiantai 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 distribution. 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 G _ST 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 990 m 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.     

企鹅珍珠贝不同地理群体遗传多样性的fAFLP 分析

为阐明企鹅珍珠贝(Pteria penguin)不同地理种群的遗传多样性机制, 采用荧光标记扩增片段长度多态性(fAFLP)技术分析了企鹅珍珠贝广西涠洲岛、广东流沙湾和海南黎安3 个不同地理群体的遗传多样性。选取7 对引物组合对90 个个体(每个群体30 个)进行fAFLP 扩增, 结果发现每个个体均能扩增出清晰的、可重复的扩增条带, 每对引物的扩增位点数在100—163 之间, 共得到895 个扩增位点, 多态位点数为865 个; 涠洲岛、流沙湾和黎安群体的多态位点比例分别为70.73%、63.13%、66.82%。Nei 遗传多样性指数为0.1634、0.1558、0.1783, Shannon 遗传多样性指数为0.2635、0.2474、0.2932。3 个群体间遗传相似度在0.9722—0.9824之间, 遗传距离在0.0177—0.0282 之间。根据遗传距离绘制UPGMA 聚类图, 但Mantel 检验结果显示企鹅珍珠贝三群体间的遗传距离与地理距离之间无显著相关。Shannon 遗传多样性指数和AMOVA 分析, 结果均显示企鹅珍珠贝的遗传变异主要来源于群体内个体间, 7.91%的遗传变异来自群体间, 92.09%的遗传变异来自群体内。分析群体的显性基因型频率分布和基因流Nm 发现3 个群体有基本相同的遗传结构, 有明显的基因交流。研究结果表明北海涠洲岛群体、湛江流沙湾群体和海南黎安群体的企鹅珍珠贝种质有较高的多态位点比例, 但未发生显著地理分化。这一结果为我国企鹅珍珠贝的良种选育以及种质资源保护措施的制定提供了参考依据。       

珍稀濒危植物金毛狗的SRAP分析

利用SRAP分子标记技术,对7个居群79株金毛狗进行遗传多样性分析。结果表明:10对SRAP引物组合共得到107条扩增条带,多态性条带比率为85.98%,Nei’s基因多样性指数为0.229 6,Shannon’s多样性指数为0.358 6,表明金毛狗居群水平具有较高的遗传多样性;金毛狗7个居群的总基因多样度为0.229 6,居群内遗传多样度为0.135 4,居群间的遗传分化指数为0.410 6,表明有41.06%的变异存在于居群间,有58.94%的变异存在于居群内;居群间基因流为0.717 8,表明居群间基因交流频率较低;遗传一致度和UPGMA聚类分析结果显示,生境条件相似的居群优先聚集,说明金毛狗种质亲缘关系与地理分布相关性不显著。     

High Genetic Differentiation of Hippophae rhamnoides ssp. yunnanensis (Elaeagnaceae), a Plant Endemic to the Qinghai-Tibet Plateau

RAPD markers were used to detect genetic diversity and population genetic differentiation of Hippophae rhamnoides ssp. yunnanensis, a sea buckthorn endemic to the Qinghai-Tibet plateau. The genetic parameters of percentage of polymorphic bands (92.86%), Nei’s gene diversity (h, 0.255), and Shannon’s index (I, 0.397) indicated high genetic diversity in this subspecies. The subpopulation differentiation suggested that 45.9% of genetic variation was among populations. High genetic differentiation among populations was also detected using AMOVA (47.02%). The main factors responsible for high genetic differentiation are probably related to natural geographic barriers among populations, gene drift, and limited gene flow caused by restricted pollen flow and seed flow. A Mantel test indicated that geographic distances were significantly correlated with genetic distances. The UPGMA phenogram based on Nei’s unbiased genetic distances and the result of three-dimensional model plots performed by principal coordinate analysis also supported the correlation. Altitude, however, did not have any clear effect on genetic differentiation.     

云南金钱槭叶子中的酚性成分

利用各种色谱技术从云南金钱槭(Dipteronia dyeriana)叶片中分离得到13个化合物。通过波谱学方法鉴定为11,12-诃子裂酸二甲酯(1),12,13-诃子裂酸二甲酯(2),11-诃子裂酸甲酯(3),12-诃子裂酸甲酯(4),13-诃子裂酸甲酯(5),鞣料云实素(6),类叶升麻苷(7),短叶苏木酚酸甲酯(8),3-O-没食子酰基莽草酸(9),樱桃苷(10),山萘酚-3-O-β-D-木糖基-(1→2)-β-D-葡萄糖苷(11),没食子酸(12),莽草酸(13)。所有化合物均首次从金钱槭属植物中分离得到。     

大猿叶虫四地理种群遗传多样性的RAPD分析

以大猿叶虫Colaphellus bowringi Baly 4个地理种群的基因组DNA为材料, 进行RAPD分析。从80条引物中筛选出11条稳定性好、多态性高的引物进行扩增, 共得到65个扩增位点, 53个多态位点, Nei氏遗传多样性指数为0.1049~0.2061, Shannon多样性指数为0.1641~0.3167。结果表明所分析的大猿叶虫遗传变异很高, 其中江西龙南种群遗传变异最小, 山东泰安种群遗传变异最高。种群间的遗传距离范围为0.0636~0.3200, 其中江西龙南种群和江西修水种群间的遗传距离最小, 哈尔滨种群与江西龙南种群间的遗传距离最大, 种群遗传距离的大小与其相对地理距离的远近吻合。结果提示种群遗传距离的大小与它们生物学上的相似性有关联。