应用AFLP技术对斜茎黄芪根瘤菌遗传多样性的分析研究

采用ＡＦＬＰ指纹图谱分析技术对来自我国不同地区的９５株斜茎黄芪根瘤菌及２４株已知根瘤菌的参比菌一起进行遗传多样性研究．结果表明,在５０％相似性水平上,全部菌株被分为３１个ＡＦＬＰ群,显示出极大的遗传多样性．相关性较高的菌株的聚类结果与先前的表型性状数值分析结果相一致．具相同ＡＦＬＰ指纹图谱的菌株都具有相同的生长速度,且其中大部分菌株来自我国同一地区,同时具很高的表型相似性．此外,本研究表明,ＡＦＬＰ指纹技术具更高的分辩率,更能揭示种内的遗传多样性     

慢生花生根瘤菌的遗传多样性研究

用ＲＥＰ１Ｒ１和ＲＥＰ２１ＤＮＡ为引物，对２２株四川土著慢生花生根瘤菌染色体ＤＮＡ的基因外回文重复序列（ＲｅｐｅｔｉｔｉｖｅＥｘｔｒｏｇｅｎｉｃｐａｌｉｎｄｒｏｍｉｃｓｅｑｕｅｎｃｅＲＥＰ）进行了ＰＣＲ扩增和凝胶电泳，并对电泳结果的指纹图型进行了相似性聚类分析．结果表明了四川土著慢生花生根瘤菌的遗传多样性．指纹图形是根瘤菌多样性研究中一种简便而有效的方法     

四川省部分豆科植物根瘤菌的遗传多样性

从四川省豆科植物鸡眼草属、合欢属、豇豆属、葛属、金合欢、杭子梢分离获得了103株根瘤菌,采用AFLP、BOXAIR-PCR研究了这些菌株的遗传多样性.结果表明,分离自四川省豆科植物6个属的根瘤菌存在明显的遗传多样性,AFLP分析中,全部供试菌株的相似性为21%,全部供试菌株可分为26个AFLP遗传群;BOXAIR-PCR聚类分析结果表明,103个菌株分在70%相似性水平处,供试菌株被分为18个BOX遗传群.除部分菌株外,多数根瘤菌按宿主类型聚群,同一宿主而地理来源不同的根瘤菌分布于不同的遗传群,表明宿主和地理环境对根瘤菌具有深刻影响.图3表1参9     

慢生型大豆根瘤菌的遗传多样性研究

采用选择性扩增片断长度多态性 (简称AFLP)DNA指纹技术对来自泰国北部的 2 45株慢生型大豆根瘤菌进行遗传多样性的研究 .通过AFLP图谱揭示出该地区的慢生型大豆根瘤菌有较显著的遗传多样性 .从 2 45株中选择出 92个代表株用计算机进行的树状图分析结果表明 ,所分析的菌株在 82 %的相似性水平上聚类成 8个群 .对这 92个代表株的部分菌株和慢生型大豆根瘤菌的参比菌株进行多聚酶链反应 (PCR)扩增的 16SrDNA的 4种限制性内切酶长度多态 (简称 16SrDNAPCR RFLP)分析 ,得出 2个不同的 16SrDNAPCR RFLP类型的菌株 ,分别与慢生型大豆根瘤菌的参比菌株Bradyrhizobiumjaponicum和Bradyrhizobiumelkanii相同 .图 1表 2参 14     

岩生植物金发草遗传多样性的ISSR和AFLP比较研究

禾本科岩生植物金发草〔Pogonatherum paniceum(Lam.)Hack.〕具有开发为新型国产草坪草种的潜力.用IS-SR与AFLP分子标记,分析了四川、重庆、云南及广西等地22个金发草种群的遗传多样性.用14个ISSR引物、10对AFLP引物分别扩增出239、485条谱带,多态位点百分率分别为94.98%、90.93%.两种标记均揭示出金发草种群具有丰富的遗传多样水平.由于标记原理不同,两种标记的结果呈现出微小的差别.相对而言,基于ISSR标记分析的遗传多样性参数高于AFLP标记得到的遗传多样性参数,这可能是由于金发草种群间存在着大量的微卫星变异所致.两种标记基于Nei遗传距离的聚类分析结果存在着一定的差异,但用Mantel测试对两种方法检测的遗传一致度进行相关性分析表明,它们之间存在着显著的相关性(r=0.3146,P=0.0150),用SPSS11.5软件进一步分析两种标记遗传一致度的相关性,Pearson相关系数为0.315(在0.01水平上),两种标记的聚类分析都揭示出金发草的遗传变异呈现出一定的地域分布规律.ISSR标记与AFLP标记均能应用于金发草种群的遗传多样性研究.图1表2参16     

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。     

中国大麦育成品种(系)的遗传多样性

为了解我国大麦育成品种(系)的遗传基础现状,利用位于大麦7个连锁群上不同位置的25对SSR引物对来自国内不同区域的大麦推广和新育成品种(系)的遗传多样性及其亲缘关系进行分析.结果表明,25对SSR引物在73个大麦品种间均有多态性扩增,每一对引物检测到的等位基因数目在2-6之间,平均为3.92个.SSR引物的多态性信息含量(PIC)变幅为0.252-0.780,平均为0.569.遗传多样性分析发现,我国不同大麦产区的皮大麦品种(系)间遗传相似性系数(GS)变化范围为0.661-0.767(平均值0.714),青藏高原地区青稞品种(系)间遗传相似性系数变化范围为0.533-0.925(平均值0.699),表明目前国内皮大麦和青稞的遗传多样性均较差、品种遗传背景单一.通过聚类分析,在遗传相似系数0.671水平处,42份皮大麦材料聚为2大类;在遗传相似系数0.618水平处,31份青稞材料聚为5大类,来源地相同的材料通常聚在同一大类或亚类,材料聚类结果与其地理来源有一定相关性.本研究表明SSR标记具有较好的遗传变异检测能力,对大麦品种(系)间的遗传多样性评价可为我国大麦育种过程中亲本材料的选择和利用提供重要依据.     

链格孢菌菌株致病性及其遗传差异性

对分离自紫茎泽兰的19个链格孢菌菌株的致病性进行了比较研究，并对这19个菌株和其它来源的3个菌株的5．8S rDNA及其两侧的ITS1区和ITS2区进行了序列分析．结果显示，链格孢菌菌株的致病性差异明显；21株链格孢菌菌株在5．8SrDNA及其两侧的转录问区（ITS区）的序列无差异，而与百日草链格孢菌（Alternaria zinnine）的序列差异显著，表明21株链格孢菌菌株属种内变异．采用AFLP分子标记技术对21株链格孢菌的DNA扩增片段长度多态性进行了分析，结果表明，链格孢菌自然变异群体内存在很高的遗传多样性．链格孢菌菌株的相似性与菌株的地理来源有一定的相关性，但基于AFLP标记划分的AFLP类群与基于寄主病情指数划分的致病力类群问关系不相一致；起始菌株与变异菌株相似性显著，但二者和致病性的关系不显著．图2表1参18     

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     

池塘浮游生物优势种组成与DNA指纹的关系

研究中国地质大学(武汉)校区3个池塘7个样点浮游生物群落的优势种组成和DNA指纹多态性.利用15种随机引物分别扩增,其中7种引物获得多态性片段.电泳共得到47条带,谱带平均多态率为96.4%.根据Nei's遗传距离矩阵构建3个池塘7个样点的遗传关系聚类树.通过优势种群组成聚类分析比较探讨浮游生物优势物种组成和DNA指纹遗传关系聚类树结构的关系.结果表明,同一池塘样点的优势种组成和DNA指纹遗传多样性相似性较高;西区池塘样点的优势种组成和遗传关系最近,其次是东区池塘的3个样点,差别最大的是北区池塘的两个样点.其原因可能是生境的隔离和差异.提示尽管池塘浮游生物优势种组成与DNA指纹结构表示的遗传关系会存在较小差异,但是分析结果基本一致.因此,利用DNA指纹技术研究池塘浮游生物生态学是可行的.     

Morphological and molecular comparisons of dominant amphioxus populations in the China Seas

The amphioxi Branchiostoma belcheri, Branchiostoma japonicum and Branchiostoma malayanum are recorded from the coast of China Seas. Six amphioxus populations comprising three Branchiostoma species collected from Hong Kong (southern China), Xiamen (south-eastern China) and Qingdao (northern China) were compared at the morphological and molecular levels. Phylogenetic separations among the species and geographic populations were evaluated by the analysis of 12S ribosomal RNA gene sequences and amplified fragment length polymorphism (AFLP) markers. From morphological characters, B. belcheri is more closely related to B. japonicum than to B. malayanum. However, phylogenetic affinities indicated by both 12S rRNA gene sequences and AFLP analysis showed a more recent phylogenetic splitting of B. belcheri and B. malayanum than that of B. belcheri and B. japonicum. In the AFLP similarity tree, geographic populations of B. japonicum were clustered into different clades. The AFLP data also showed that both B. belcheri and B. japonicum populations in Hong Kong have the highest levels of within-population genetic diversity as compared to that in Xiamen and Qingdao, suggesting that genetic diversity of Branchiostoma decreases from low to high latitudes. Results of hierarchical analysis of molecular variance (AMOVA) also revealed a high level of genetic diversity either for the three B. japonicum populations or the two B. belcheri populations in China Seas. However, genetic variation among the three B. japonicum populations was insignificant, indicating that these populations are genetically connected.     

An AFLP-based test of clonality in widespread, putatively asexual populations of Porphyra umbilicalis (Rhodophyta) in the Northwest Atlantic with an in silico analysis for bacterial contamination

Asexual reproduction often predominates in populations of species experiencing range expansion. Locally selected genotypes can lead to the establishment of clonal populations that accumulate genetic diversity through time. Sexual reproduction has never been observed in extensive field and culture studies of the red alga Porphyra umbilicalis from Maine, USA, even though sexual reproduction predominates in this species in the eastern Atlantic (Europe). This suggests that Maine populations are indeed asexual and might consist of one or only a few genetic clones; we have tested this using AFLPs. Individuals were sampled at two sites in Maine (Cobscook Bay [n?=?25], Schoodic Point [n?=?26]) and compared to sexual individuals from England (Sidmouth [n?=?17]). The AFLP analysis determined that individuals at two sites in Maine containing putative asexuals were not strictly clonal; however, two multilocus lineages were sampled more than once. Two genetic clones, one at each Maine site, were comprised of 6 individuals each; the 39 additional Maine individuals had distinctive AFLP genotypes. However, when the individuals from Maine were compared with a known sexual population from Sidmouth, England, much greater genetic diversity was found within the sexual population in England. Finally, we examine how preparation of field-collected material for AFLP investigations can affect the inclusion of non-target DNA and demonstrate an in silico approach for removing some cryptic contaminants from analysis.     

Limited population structure in Northern and Spotted Wolffishes (<Emphasis Type="Italic">Anarhichas denticulatus</Emphasis> and <Emphasis Type="Italic">A. minor</Emphasis>) despite low apparent dispersal potential

Northern and Spotted Wolffishes (Anarhichas denticulatus and A. minor) are demersal marine fishes listed as “threatened” in Canadian waters. Both species have unusually large benthic eggs and large size at hatch, which should reduce passive dispersal. We examined population differentiation with microsatellite and AFLP loci across the ranges of both species in the North Atlantic Ocean. Although significant population structure was documented, differentiation was less than expected based on knowledge of life history characteristics. Significant differentiation was found in Northern Wolffish between the Barents Sea and other samples based on both microsatellite and AFLP data. In contrast, population structure in the Spotted Wolffish was notably weaker, particularly with microsatellites. Both species were characterized by low genetic diversity for marine fishes and had significantly lower genetic diversity than the congeneric Atlantic Wolffish. This finding was consistent with the conservation status of these three species and suggests potential vulnerability to over-exploitation in Northern and Spotted Wolffishes.     

潍坊、花园口土样中土著大豆根瘤菌的遗传多样性研究

采用１６Ｓ－２３Ｓ ｒＤＮＡ间隔区段（ＩＧＳ）ＲＦＬＰ分析和ＰＡＰＤ分析技术，分别对分离自山东潍坊和河南郑州花园口的２４株土著大豆根瘤菌进行２多样性分析。结果一 ７０％的相似性水平上，依ＩＧＳ－ＲＦＬＰ分析可将供试菌株分为１０群，依ＲＡＰＤ分析可将供试菌株分为８群。综合两种分析技术在５０５的相似性水平上将供试菌株分为潍坊群和花园口群。 在系统发育和分子进行上有明显的地理分隔作用。     

Multigenerational genetic analysis of tropical secondary regeneration in a canopy palm

Iriartea deltoidea (Arecaceae) is an abundant canopy palm with a wide geographic distribution in Neotropical wet forests. We analyzed the genetic profile across three generations of Iriartea within a 43-ha area encompassing two areas of second-growth and adjoining old-growth forest at La Selva Biological Field Station in northeastern Costa Rica. A total of 311 reproductively mature trees, 99 large saplings, 207 small saplings, and 601 seedlings were genotyped using 141 AFLP loci. Parentage analysis revealed high dispersal distances, both for seed (over 2.3 km) and pollen (over 3.8 km), indicating a large genetic neighborhood within La Selva Biological Station. In a 20-ha area of second growth, the founding palm population was dominated by a small number of parental trees located in the adjacent old-growth forest; two old-growth trees contributed 48% of the second-growth genes. The genetic diversity of reproductively mature trees in this second-growth forest was significantly reduced compared to adjacent old-growth forest. Within 400 m of the border with old-growth forest, we observed a similar reduction of genetic diversity in saplings, and an even greater loss of genetic diversity in the second generation of seedlings. Nearly half of these seedlings were offspring of local parents. In contrast, in the distant portion of second-growth forest (400-800 m from the old-growth border), parentage analysis showed that 40% of seedlings originated from outside the study area and only 10% were offspring of local parents. These high levels of gene flow maintained genetic diversity in saplings and seedlings similar to levels observed in old-growth forest. Our findings highlight the importance of gene flow from diverse seed and pollen sources for sustaining levels of genetic diversity of tree populations in second-growth forests.