皮下盘菌属种内及种间遗传多样性的ISSR分子标记

对皮下盘菌属（Hypoderma）4种23个菌株进行了ISSR分析。结果表明,被试材料ISSR标记多态性好,8个引物共扩增出131个条带,其中126条（占9618％）具有多态性。经欧氏最短距离中的类平均法聚类,得到各菌株之间的ISSR标记的遗传相似性系数（GS）变异范围为0．30—0．82,23个菌株可聚为6类。通过ISSR分析结果与表型性状的比较和分析,明确了悬钩子皮下盘菌种内遗传差异体现在子囊果大小、形状及埋生深度、唇特征、侧丝顶端形状、子囊和子囊孢子形状及大小,以及寄主、着生部位和分布区域方面;皮下盘菌属的种间差异主要表现在子囊果、唇、侧丝、子囊及分生孢子器等部分形态学特征、寄主及着生部位方面。     

安徽野生香菇遗传多样性及杂种优势的ISSR分析

采用简单序列重复区间扩增多态性(Inter–simplesequencerepeat,ISSR)技术,利用13个引物对26个安徽野生香菇Lentinulaedodes菌株和6个香菇栽培品种进行了遗传多样性分析,构建了遗传相关聚类图,并根据ISSR分析选择遗传距离远近不同的亲本进行组合杂交,评价了其杂种优势。在78个ISSR标记中,多态性标记为61个,占78.2%。聚类分析显示,当以相异距离0.23为阈值,32个菌株被划为5个ISSR遗传组,多数菌株之间遗传相似性较低。这表明供试菌株在DNA水平上存在比较显著的遗传变异,具有较丰富的遗传多样性。杂种优势的检测表明亲本间遗传距离较大的组合其杂种优势也较强。     

弯孢类炭疽菌菌株的RAPD分析与分类研究

通过RAPD分析对38个不同寄主来源的弯孢类炭疽菌菌株的系统发育及分类进行研究。结果表明:许多引物的RAPD扩增带型在种内是相似或一致的,而种间差异较大。UPGMA聚类分析的结果表明:38个弯孢类炭疽菌菌株被聚为6个类群,群内菌株之间遗传相似性较高,而群间遗传相似性较低;群与群之间分界明显,表明种的分界相当明显。因此,RAPD分析所反映的种间亲缘关系,将有助于疑难种分类地位的确定和近似种的区分。RAPD分析的结果还揭示了一些近似种的分类关系,如按传统方法分别鉴定为Colletotrichumtruncatum、C.circinans和辣椒炭疽菌C.capsici的许多菌株聚类在同一群内,有很高的遗传相似性,实验结果支持它们为同一个种。实验中还发现5条辣椒炭疽菌的特有片断,分别由引物OPE-14、OPH-15(2条)、OPM-12和OPM-20扩增产生。这些特异带的发现对该菌的快速检测和鉴定具有重要意义。     

22种木莲属植物亲缘关系的ISSR分析

利用ISSR标记分析技术探讨了22种木莲属植物的遗传亲缘关系。选用10对ISSR引物组合对木莲属22个种的基因组进行分析。结果表明：22个种共扩增出595条DNA带,其中多态性条带占94.45%;根据Nei Li遗传相似性系数在0.68处,UPGMA聚类结果将22个种划分为6个类群,在0.70处进一步可分为12个亚类群;22种木莲属植物的Nei-Li遗传相似性系数变化范围为0.605(乳源木莲与球果木莲之间)～0.956(中缅木莲与滇南木莲之间),平均遗传相似性系数为0.698。说明木莲属植物种间差异性大。其中中缅木莲、滇南木莲之间遗传相似性系数为0.956,两者亲缘关系最近。该结果同时支持将巴东木莲、乳源木莲、滇桂木莲分别作为独立的种。     

被子植物生散斑壳属若干种内和种间亲缘关系RAPD分析

采用9个重复性好的随机引物对被子植物上的28个散斑壳属(Lophoderm iumChevall.)菌株进行RAPD-PCR扩增,通过RAPD分析结果结合表型性状的比较和分析,确定了子囊果埋生位置,子座、子囊、子囊孢子、侧丝等形态学特征以及寄主种类差异在该类菌物种水平分类上的重要性,另外产地不同等对种内遗传多样性产生一定影响。进一步明确了被子植物生散斑壳属一些种内和种间的亲缘关系。     

重要昆虫病原真菌粉棒束孢种群的遗传分化

粉棒束孢是一种重要的昆虫病原真菌.运用ISSR分子标记研究了安徽省6个不同地理环境的粉棒束孢种群遗传异质性.结果表明:10个多态性引物多态位点百分率高达98.5％,但种群水平的多态位点差异较大,在59.6％ - 93.2％.基于Nei遗传异质性分析得出各种群间的遗传分化系数(Gst)为0.3365,基因流(Nm)为0.4931;各种群间的遗传分化低于种群内的遗传分化,表明安徽省粉棒束孢的遗传变异主要存在于种群内.根据各菌株间的遗传相似系数进行UPGMA聚类,结果表明,西山种群是单系的同质种群,其余5个种群皆为多系的异质种群,其中鹞落坪种群的异质性最高,琅琊山种群异质性最低.各种群之间的地理距离与遗传距离之间无相关关系.根据6个种群间的遗传距离进行UPGMA聚类,可将它们分成3个类群,分类的结果与各种群的地理环境相符,反映出环境的异质性对种群异质性的影响.     

用ISSR标记检测黄麻野生种与栽培种遗传多样性

利用ISSR标记对黄麻属((Corchorus)10个种27份材料的遗传多样性进行分析,25个ISSR引物共扩增出283条带,平均每个引物扩增出10．48条带,多态性条带比例(PPB)为92．85％;种间遗传相似系数在0．33～0．97之间。表现出丰富的遗传多样性．系统聚类结果显示,第I、Ⅲ类群均为黄麻属8个种11份原始野生种,种间存在丰富的遗传多样性;第Ⅱ类群为黄麻两个栽培种及其近缘野生种,共16份材料。种遗传相似性较高;基因组聚类结果与经典分类相符．利用分子标记技术研究初步可以认为。荨麻叶种为最原始的黄麻野生种之一;三室种21C为三室种的一个变种;甜麻为一个尚待定性的野生种．同组材料的RAPD和ISSR分析结果,具有较高的相似性和可信度．     

五种不同蛏类ITS2核苷酸序列分析

对5种不同蛏类的ITS2核苷酸序列进行扩增、测序及分析.结果表明:5种蛏类的509个序列位点中共有49个保守住点,402个变异位点,318个简约信息位点,84个单突变位点.5种不同蛏类遗传距离在1.194～3.173之间,总角截蛏和小刀蛏的遗传距离较近,而与其他3种蛏类的遗传距离较远,遗传距离在1.965～3.173之间.缢蛏、长竹蛏和细长竹蛏3种蛏之间的遗传距离较其他2种蛏近,遗传距离在1.194～1.318之间.同种蛏类不同个体之间的遗传距离很近,遗传差异很小.     

内蒙古地区刺叶柄棘豆(Oxytropis aciphylla Ledeb.)种群遗传多样性分析

用ISSR分子标记对内蒙古地区刺叶柄棘豆(Oxytorpis aciphylla Ledeb.) 5个地理种群进行了种群遗传多样性分析。结果表明：刺叶柄棘豆种群具有较高的遗传多样性,11个ISSR引物扩增出215条带,总的多态位点百分率为98.14%,Shannon多样性指数I=0.2108,Nei基因多样性指数 H=0.341 6,种内总基因多样性(H_t) 为0.2108,种群内基因多样性(H_s)为0.160 4, 大部分遗传变异(76.11%)的遗传变异存在于种群内, 23.89%的遗传变异存在于种群间。遗传分化系数(Gst)为0.238 9,基因流(Nm)为1.592 9。5个居群间已有遗传分化趋势,遗传漂变不会引起遗传分化。UPGMA遗传距离聚类结果表明, 5个地理种群中,植被类型为荒漠草原的4个种群之间遗传距离较近,与1个荒漠种群距离较远。     

广东省水稻纹枯病菌遗传多样性与致病力分化的研究

利用随机引物扩增多态性DNA（PAPD）分子标记技术分析了来自广东省7个县市48个水稻纹枯病菌菌株的遗传多样性,以筛选出的10个随机引物对菌株进行PAPD－PCR扩增,共产生了98个PAPD分子标记,其中89．9％的片段具有多态性。48菌株间的遗传相似性系数（以Nei基因一致度表示）在0．56－0．949之间,用UPGMA和类分析可将它们分为5个RAPD遗传聚类群（A、B、C、D、E）,相同地区来源的菌株基本上聚类在同一组群内。在温室中对供试菌株进行致病性测定,结果表明所有菌株对水稻品种Tetep都有致病性,菌株间致病力差异显著（α＝0．05）,病情指数范围为0．73－18．7,平均感指病指数为5．24。试验分析结果表明水稻纹枯病菌具有丰富的遗传多样性,不同县市的菌株存在很大的遗传分化现象（FST＝0．579）,RAPD遗传聚类群的划分与菌株的地理来源有明显的相关性,但菌株的致病力差异与菌株的来源、遗传聚类组群的划分没有明显的相关性。     

Analyzing genetic relationships in Bombyx mori using intersimple sequence repeat amplification

Intersimple sequence repeat (ISSR) amplification was used to analyze genetic relationships among silkworm, Bombyx mori L., strains. Nineteen primers containing simple sequence repeat (SSR) motifs were tested for amplification on a panel of 42 strains, representative of the diversity of silkworm germplasm; 12 of the primers amplified distinct, reproducible bands. The primers amplified a total of 108 bands, of which 85 (78.7%) were polymorphic. The ISSR results suggested that within the dinucleotide class, the poly(CA) motif was more common than the poly(CT) motif. The ISSR amplification pattern was used to group the silkworm strains into seven subclusters based on their origin in an unweighted pair-group method with arithmetic average cluster analysis by using Nei's genetic distance. Seven major ecotypic silkworm groups were analyzed. Principal component analysis of the ISSR data supported the unweighted pair-group method with arithmetic average clustering. Therefore, ISSR amplification is a valuable method for determining genetic variability among silkworm varieties. This efficient genetic fingerprinting technique should be useful for characterizing the large numbers of silkworm strains held in national and international germplasm centers.     

杏鲍菇菌种遗传多态性的ISSR分析

采用ISSR(Inter-simple Sequence Repeat)分子标记技术并应用NTSYSpc 2.10e生物软件对20株杏鲍菇菌种进行遗传聚类分析。结果表明,从22条引物中筛选出11条扩增条带清晰、多态性好的ISSR引物,共获得74个ISSR标记位点,51个多态性位点,多态比率为68.92%,产物片段大小在100~5 000 bp之间。聚类分析发现,遗传相似系数在0.828时,20株杏鲍菇菌种分为4个类群,当遗传相似系数达到0.896时,分为12个类群,杏528寿与其他菌种的亲缘系数最远,具有独立的遗传体系。     

羊肚菌不同单孢及不同萌发孔菌丝间的遗传多样性

本研究对来自不同单孢和不同萌发孔菌丝的166份羊肚菌菌株进行形态研究、交配型基因检测、基于ISSR的遗传多样性分析和菌株杂交选育。结果表明,来自不同单孢及其不同萌发孔菌丝培养的菌株间均存在不同程度的形态和遗传差异。4条ISSR引物共扩增出条带清晰的22条多态性谱带,多态性比率为88%。基于遗传相似性系数（GS）、不加权成对群算术平均法（UPGMA）构建的系统树,可将供试菌株分为梯棱羊肚菌Morchella importuna和六妹羊肚菌M. sextelata两大类群。聚类分析发现不同物种间的单孢菌株和单丝菌株的遗传差异显著;同一单孢不同萌发孔菌丝的菌株间存在较大的遗传分化,遗传多样性丰富。交配型基因检测证实羊肚菌是异宗结合型真菌,同一单孢的不同萌发孔菌丝体含相同的交配型基因（MAT 1-1-1或MAT 1-2-1）。遗传多样性结果表明：梯棱羊肚菌比六妹羊肚菌的遗传背景更广泛。此外,ISSR分子标记也表明仅依靠菌丝和菌核的形态特征并不能准确衡量羊肚菌菌株间的亲缘关系。研究羊肚菌单孢菌株、单丝菌株间的形态和遗传特征,将为羊肚菌优质菌种的精准选育提供理论参考。     

Genetic diversity of wild Auricularia auricula-judae revealed by ISSR analysis

Auricularia auricula-judae is an edible and medicinal jelly mushroom, and it is widely cultivated in China. In the present study, inter-simple sequence repeat (ISSR) molecular marker was employed to assess the genetic diversity of 22 wild strains and two cultivars of A. auricula-judae obtained from many ecological regions of China. With the use of 11 ISSR primers, a total of 368 (99.7%) and 81 (71.7%) polymorphic loci in wild and cultivated strains were detected, respectively. The mean genetic similarity of A. auricula-judae was 0.75, based on the dendrogram generated via similarity coefficient, all the tested strains were classified into six groups at a similarity level of 76%, and the two cultivated strains consisted of one group alone. The results of principal coordinate analysis were in accordance with UPGMA clustering, and the first most informative coordinate accounted for 75.9% of the variations, indicated a high level of genetic diversity among wild A. auricula-judae strains. In general, ISSR marker was an effective method to discriminate the A. auricula-judae stains and to evaluate their genetic diversity.     

21株马特组镰刀菌遗传多样性的ISSR分析

为明确马特组镰刀菌种间和种内的遗传差异与亲缘关系,本文利用ISSR分子标记技术对21个马特组菌株进行了遗传多样性分析.结果表明:利用筛选出的15条引物对3种供试菌株进行扩增,共扩增出239条条带,其中多态性条带230条,多态性位点比例为96.2％,平均每条引物产生条带数为15.3条.21个菌株间的遗传相似系数范围为0.494～0.933,平均为0.640.在遗传相似系数为0.593时,供试的21株马特组镰刀菌可明显分成2个ISSR类群(IG),IG-Ⅰ包括1～17号菌株,为Fusarium solani和F solani var.coeruleum;IG-Ⅱ包括18 ～21号菌株,全部为F.ventricosum.在遗传相似系数为0.933时,供试的21个菌株可被全部区分开.供试的镰刀菌基因组在SSR区域具有丰富的多态性;ISSR类群划分与菌种分类之间存在一定相关性,但与菌株的地理来源没有相关性;而同一类群中,不同菌株之间的遗传相似性与菌株的地理来源存在一定的相关性.同一地区同种寄主的相同菌种,其菌株间也存在一定的遗传差异.     

Differentiation of commercial strains of <Emphasis Type="Italic">Agaricus</Emphasis> species in China with inter-simple sequence repeat marker

In recent years, Agaricus mushroom production has undergone an important increase in the world. However, there is no consensus over the taxonomy of some commercial strains. So rapid and reliable methods to identify Agaricus strains are urgently needed. In this paper, six inter-simple sequence repeat (ISSR) primers were screened from 20 primers to analyse the genetic diversities of the 12 main commercial strains of Agaricus in China. The results showed that a total of 124 bands were produced, among which 80.6% was polymorphic, the similarity coefficients ranged from 0.69 to 0.98. Twelve tested strains were divided into 2 groups on the similarity coefficient of 0.75. Three Agaricus bitorquis strains were clustered in one group, and seven Agaricus bisporus strains were clustered with Agaricus strains S1 and S2 in the other group. As comparison with ISSR, intergenic spacers combined with restriction fragment length polymorphisms (IGS-RFLP) was employed to analyse genetic diversities of the tested strains by digesting IGS amplified products with restriction enzymes Hae III, Alu I, Rsa I, respectively, and IGS-RFLP cluster results were similar with those of ISSR. However, ISSR was superior to IGS-RFLP in differentiating closely related strains. It is suggested that the ISSR marker is an alternative method to differentiate Agaricus strains.     

Evaluation of Genetic Diversity Among Chinese Pleurotus eryngii Cultivars by Combined RAPD/ISSR Marker

Pleurotus eryngii (DC. Ex. Fr.) Quél is a rare precious edible fungus which belongs to the family Pleurotaceae. This mushroom has highly nutritional, pharmaceutical, economic and ecological values. In the present study, combined randomly amplified polymorphic DNA (RAPD)/inter-simple sequence repeat (ISSR) was used to assess the genetic diversity of P. eryngii strains cultivated in China. For the RAPD and ISSR analyses, 404 and 392 polymorphic bands were obtained from 32 P. eryngii strains using 28 and 24 selected primers, respectively. A combined RAPD/ISSR dendrogram grouped the 32 strains into five clades with coefficient of 0.770. The comparison of RAPD and ISSR was also elucidated in the present study. The results of our study obtained by combined RAPD/ISSR analysis contributed to a better understanding of the genetic relationships among the P. eryngii strains and provide orientation for the strain improvement of P. eryngii species.     

Analysis of genetic relationship in mutant silkworm strains of Bombyx mori using inter simple sequence repeat （ISSR） markers

Amplified inter simple sequence repeats (ISSR) markers were used to determine genetic relationships among mutant silkworm strains of Bombyx mori. Fifteen ISSR primers containing simple sequence repeat (SSR) motifs were used in this study. A total of 113 markers were produced among 20 mutant swains, of which 73.45% were found to be polymorphic. In selected mutant genetic stocks, the average number of observed allele was (1.7080±0.4567), effective alleles (1.5194±0.3950) and genetic diversity (Ht) (0.2901±0.0415). The dendrogram produced using the unweighted pair group method with arithmetic means (UPGMA) and cluster analysis made using Nei's genetic distance resulted in the formation of one major group containing 6 groups separated 20 mutant silkworm strains. Therefore, ISSR amplification is a valuable method for determining the genetic variability among mutant silkworm swains. This efficient molecular marker would be useful for characterizing a considerable number of silkworm swains maintained at the germplasm center.     

57株毛头鬼伞遗传多样性分析

运用SRAP、RAPD、ISSR3种分子标记技术对来源不同地区的57株毛头鬼伞Coprinus comatus进行了遗传多样性分析,通过3种分子标记进行聚类分析,当相异系数D为0.48时,可以把57株毛头鬼伞分为4类:Ⅰ类包括Co0001;Ⅱ类包括Co0003;Ⅲ类包括Co0005;Ⅳ类包括其余54个菌种。供试的57个菌株间的相异系数范围从0–0.72,具有一定的遗传多态性。但其中有许多菌株两者之间的相异系数为0,说明毛头鬼伞菌种存在着比较严重的同种异名现象。