65份野生油茶种质遗传多样性的ISSR和RAPD标记分析

利用ISSR和RAPD标记,对名邛台地野生油茶种质进行遗传多样性分析。从60条简单重复序列引物中筛选出16条引物,在65份样品中共扩增出213条带,其中多态位点为203个,多态位点百分率为95.31%;从30条寡居核苷酸引物中筛选出8条引物,共扩增出105条带,其中多态性位点94个,多态位点百分率为89.52%。结果表明:名邛台地野生油茶种质具有较丰富的遗传多样性,ISSR和RAPD标记可以应用于油茶种质遗传多样性分析。     

野生罗汉果遗传多样性的ISSR分析

应用ISSR分子标记方法对采自广西和广东的7个罗汉果（Siraitia grosvenorii）野生居群共130个个体进行了遗传多样性分析。15个ISSR引物共扩增到了111个位点,其中91个是多态性位点,占82.0%。Nei′s基因多样性指数(H_e)为 0.248,Shannon 信息多样性指数（I） 为0.354。罗汉果不同居群的遗传多样性水平差异较大,居群多态位点百分率在 28.2%－55.6%之间,Nei′s基因多样性指数为0.080－0.209,Shannon 信息多样性指数为0.123－0.310。永福居群（YF）和金秀居群（JX）的遗传多样性水平较高,其周边居群的遗传多样性水平逐渐降低,居群间产生了较大的遗传分化（G_st = 0.569）。居群间的遗传距离与地理距离相关性不明显（r =0.369,P = 0.115）。UPGMA聚类图中,7个居群的个体按居群各自聚在一起。     

河南大豆遗传多样性的ISSR分析

用ISSR标记技术对10个大豆品种进行遗传多样性分析.从44条随机引物中筛选出8个多态性引物,共扩增出89条带,其中有55条为多态性条带,多态性比率为61.8%,条带大小为220~1 500 bp,平均每个引物可扩增出11条带.Shannon多样性指数评价结果表明,平均多样性指数为0.286 5,观察等位基因数和有效等位基因数分别为1.618和1.300 8;统计分析结果表明,10个品种间的相似系数为0.60~0.75,平均相似系数为0.69;聚类分析结果表明,10个大豆品种可聚成2组:第一组包括豫豆15、豫豆11、豫豆24、周豆12和周豆11;第二组包括豫豆22、周豆13、豫豆6、中作98-3和豫豆26;主成分分析结果支持聚类分析结果.本研究为大豆品种鉴定和种质资源利用奠定了基础.     

云杉矮槲寄生遗传多样性的ISSR分析

为探究云杉矮槲寄生(Arceuthobium sichuanense)的遗传多样性及其与不同寄主选择压力和地理分布的关系,采用ISSR分子标记方法,对青海、甘肃、四川等地区5种寄主上的100份云杉矮槲寄生样本进行遗传多样性和遗传结构分析。结果表明:(1)10条引物共扩增出130个条带,其中多态性条带129条,多态位点百分率(PPB)为99.23%。(2)物种水平上的Nei’s遗传多样性指数(He)和Shannon信息指数(I)分别为0.313 9和0.476 5,表明云杉矮槲寄生物种水平的遗传多样性较高,但群体间的基因流(Nm=0.528 7)较弱,可能会加速群体间的遗传分化(Gst=0.486)。(3)UPGMA聚类结果显示,来自甘肃、青海的样本聚为一组,表现出较高的相似性,而四川的样本独立聚类;不同寄主来源的聚类结果显示,寄生于鳞皮云杉(Picea asperata)与川西云杉(P.likiangensis var.balfouriana)的样本聚为一类,而寄生于青海云杉(P.crassifolia)、青杄(P.wilsonii)和紫果云杉(P.purpurea)的样本聚为一类,表明地理隔离和寄主选择压力对云杉矮槲寄生的遗传分化起到了重要作用。     

黄芩种质资源ISSR遗传多样性的分析及评价

采用ISSR分子标记技术对6个野生或栽培居群共147份黄芩种质进行遗传多样性分析和评价。分析结果表明,51个ISSR引物中筛选出18条扩增条带清晰、重复性好和多态性高的引物,共扩增出485条清晰的条带,其中466条具有多态性,平均多态性位点比率为96.08%,平均Nei’s基因多样性指数和Shannon’s信息指数分别为0.244 4和0.388 9,等位基因数（Na）和有效等位基因数（Ne）分别为1.993 8和1.383 9,遗传分化指数G_st=0.122 3,遗传一致度（I）和遗传距离（D）分别为0.951 5和0.050 1,说明收集的黄芩种质资源在总体上具有较高的遗传多样性,不同居群间存在一定的遗传分化和基因交流,遗传变异主要存在于居群内。分子聚类结果表明,同一地区的种质并没有按照收集来源完全聚类,可能与种质不同起源或民间栽培引种有关。在DNA分子水平揭示黄芩种质资源的遗传多样性水平,将为进一步黄芩种质资源评价、保存和新品种选育等利用提供依据。     

舟山群岛野生山茶种群遗传多样性的ISSR分析

采用ISSR分子标记,利用筛选的20条引物对舟山群岛4个野生山茶（Camellia japonica）种群的遗传多样性进行分析。结果表明：山茶种群的多态位点百分比（PPB）为64.06％,Nei’s基因多样性指数（HE）为0.2390,Shannon信息多态性指数（H）为0.3548,种群水平遗传多样性较高。基因分化系数Gst = 0.2241,表明种群间具有较高的遗传分化。地理距离与遗传距离具有显著相关性（r = 0.9653,P<0.05）,表明岛屿隔离对山茶种群的遗传分化具有重要影响。UPGMA聚类表明同岛种群间的亲缘关系更近。基于舟山群岛山茶种群遗传结构分析,建议在其自然生长地加强就地保护。     

湖北野生春兰资源遗传多样性的ISSR分析

近年来,由于过度采挖和生境片断化,湖北野生春兰(Cymbidiumgoeringii)资源正面临着灭绝的危险。本文采用ISSR分子标记技术对湖北省内的11个春兰野生居群共325个个体的遗传多样性水平及居群遗传结构进行了研究。11个引物共检测到127个位点,其中112个为多态位点,占88.19%。POPGENE分析结果表明:与其他兰科植物相比,春兰具有丰富的遗传变异(在物种水平上,He=0.2628,Ho=0.4037;在居群水平上,PPL=63.06%,He=0.1945,Ho=0.2958)。Nei's遗传多样性分析和AMOVA分析表明,各居群间产生了一定程度的遗传分化(GST=0.2440,FST=0.2207)。居群间一定程度的遗传分化可能是由生境破坏和基因流障碍(Nm=0.8828)引起。UPGMA聚类分析可知,与其他居群相比,恩施地区的5个居群,即巴东(BD)、福宝山(FBS)、宣恩(XE)、毛坝(MB)、来凤(LF)优先聚成一支,而大悟(DW)居群单独聚为一支。同时本研究也表明,虽然春兰自交亲和,但在自然界中其繁育系统还是以异交为主。鉴于春兰资源的遗传多样性现状和其相应的居群遗传结构,我们建议在遗传多样性较高的来凤(LF)、京山(JS)、大悟(DW)居群设立保护点进行就地保护;而对资源破坏最为严重的毛坝(MB)和宣恩(XE)居群要实行迁地保护。     

华南及邻近地区大刺鳅遗传多样性的ISSR分析

为了解我国大刺鳅野生资源状况, 研究采用ISSR技术分析了福建、广东、广西、云南和海南11个地理群体262尾大刺鳅的遗传多样性。10个引物共扩增出112个位点, 多态位点95个, 多态位点比例为84.82%。大刺鳅总群体Nei基因多样性h=0.2126、Shannon 信息指数I=0.3358, 表明大刺鳅总群体遗传多样性较丰富, 其中各群体遗传多样性依次为恩平 红河 屯昌 英德 河池 乐昌 增城 龙岩 五华 仁化 百色。总群体遗传分化系数Gst=0.4620, 显示46.2%的变异来自群体间。总群体基因流Nm=0.5823, 表明大刺鳅总群体间缺乏有效的基因交流, 遗传漂变是大刺鳅群体遗传分化的主要因素。聚类分析表明, 东江群体和韩江群体聚为一支, 西江群体和北江群体聚为一支, 大陆群体聚为一大支, 然后和海南屯昌群体聚类。     

观叶福禄桐遗传多样性的ISSR分析

利用ISSR分子标记对9个观叶福禄桐品种进行遗传多样性和亲缘关系分析,从100条引物中筛选出9条稳定、多态性高的引物用于PCR扩增,共获得70条带,其中多态性条带61条,多态百分率为87.14%。聚类结果显示,品种间相似性系数为0.346 9-0.816 3,聚类结果与品种间的地理来源紧密相关,从外观形态上比较,亲缘性较近的叶形和株型相似性较高;观叶福禄桐各品种间基因型差异较小,亲缘关系较近,遗传基础相对较窄。     

用ISSR标记技术分析山药品种遗传多样性

利用ISSR标记技术对28个山药品种的遗传多样性进行分析。结果表明,从44条ISSR引物中可筛选出7条能够扩增出清晰、稳定条带的引物;这7条ISSR引物对28个山药品种扩增条带间存在较大差异,多态性条带比率为83．01％,Shannon多样性指数为0．3191;构建的分子树状图将28个山药品种划分为4组：第一组含有日本白、花山药和日本园3个品种;第二组为小叶山药;第三组为嵩野1号;其余23个品种归入第四组。而且主成分分析结果支持上述的聚类分析结果。这为利用ISSR标记技术鉴定山药品种,为有效地利用山药种质资源提供了依据。     

Evaluation of genetic diversity in Lentinula edodes strains using RAPD, ISSR and SRAP markers

Random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) markers were used to evaluate the genetic diversity among 23 elite Lentinula edodes strains in China. A total of 138, 77 and 144 bands were detected by 16 RAPD primers, 5 ISSR primers and 23 SRAP primer combinations, among which 58.8%, 73.5% and 56.3% was polymorphic, respectively. By UPGMA clustering, a dendrogram was constructed based on each analysis. The three dendrograms showed that 23 L. edodes strains were clustered into three or four groups. The grouping exhibited similar structure and was generally consistent with their pedigrees. Twenty-three L. edodes strains shared great similarity indicated that the low level of genetic diversity of L. edodes strains and their relationship between each other. The important source of breeding material, such as wild and exotic types, must be introduced in order to broaden genetic base and decreases genetic vulnerability of L. edodes.     

香菇单孢杂交子代群体灰色关联度和ISSR分析

以香菇Lentinula edodes栽培菌株秋6和K95-1为亲本,通过单孢菌株配对杂交获得21个杂交子,观察杂交子及亲本菌丝体生长情况,进行栽培出菇试验。采用灰色关联度分析法,对9个正常出菇的杂交子从8个性状方面进行综合评价,结果表明,杂交子QK-8和QK-15关联度仅次于亲本秋6,农艺性状和子实体性状表现最好。采用ISSR技术对21个杂交子及亲本进行DNA多态性聚类分析,单孢杂交后代遗传分化十分明显,归于同一类群的杂交子在菌丝生长、子实体形态和农艺性状等方面常表现相似,ISSR分析能为优良杂交子初步筛选提供重要参考。     

香菇主要栽培菌株遗传多样性的AFLP分析

采用AFLP技术分析了收集到的31个主要香菇栽培菌株的DNA多态性。采用6对引物共扩增得到了443条DNA带,其中共有条带为189条,多态性比率为57.34%,说明收集到的香菇菌种具有一定的遗传多样性,它们之间存在一定的遗传差异。用平均连锁聚类法构建了所有样本的遗传相关聚类图,以0.80的相似性为切割点,31个菌株可分成4个类群,类群Ⅰ主要由上海农科院食用菌研究所和福建三明真菌研究所提供的香菇菌种组成,类群Ⅱ全部由收集到的日本栽培品种组成,类群Ⅲ由浙江庆元与福建三明真菌所的菌种组成,类群Ⅳ为上海农科院食用菌所的7402。本研究为香菇遗传信息数据库的建立奠定基础,为优良品种选育和亲缘关系的研究提供分子生物学依据。AFLP技术通过不同菌株的指纹图谱的不同能够有效分辨其基因型,可为香菇的栽培菌株质量监测和菌种鉴定提供快速有效的技术手段,从而为规范食用菌菌种生产管理提供科学依据。     

用ITS和ISSR分子标记技术鉴别香菇生产用种

通过选用香菇生产中存在名称争议或者名称相近或者同一名称但长期在不同地区栽培的12株香菇生产菌株,以及用于种水平对比的豹皮香菇Lentinuslepideus和虎皮香菇Lentinustigrinus的4个菌株,共16个菌株作为供试材料,进行ITS和ISSR遗传分析,并用RAPD技术验证试验结果。结果证明,不同种的ITS长度存在差异,再次证明ITS可以有效区别种之间的菌株;在ISSR的菌株水平分析中,香菇种内材料拥有两个共同的条带,与其他两种菌株的带型图谱有着明显差异,其中5对材料的带型图谱极为相近。RAPD验证结果与上述结果相近。由此可见,结合ITS与ISSR技术是可以用作香菇生产菌株鉴别的,这为ITS和ISSR分子标记技术推广应用于香菇生产菌株的快速准确鉴别提供了技术依据。     

Using SSR markers to evaluate the genetic diversity of Lentinula edodes’ natural germplasm in China

In this study, microsatellite markers were utilized to reveal the genetic diversity of 56 strains of Lentinula edodes grown in China. A total of 224 DNA bands were detected through 25 primer pairs, of which, 223 bands (99.6%) were polymorphic between two or more strains. The variation in SSR (Simple Sequence Repeat) DNA band patterns and average genetic similarity among the wild strains of L. edodes obtained from the same region uncovered a vast genetic diversity in the natural germplasm found in China. Compared with L. edodes strains from other areas, the genetic diversity of those from the Yunnan Plateau, Hengduanshan mountains, Taiwan, and south China was significantly greater. Based on cluster analysis and principal coordinate analysis, the results indicated that all L. edodes strains could be divided into three major groups. These results effectively displayed the differences between the strains from North and South China, and those from the same or adjoining regions could cluster preferentially into small groups in most cases, suggesting the positive correlation between the cluster results and the geographical origin for the natural germplasm of Chinese L. edodes. To our knowledge, this is a pioneering report on the utilization of the SSR marker technique in analyzing L. edodes’ genetic diversity.     

Assessment of the genetic diversity and genetic relationships of Lilium in China using ISSR markers

China is a center of natural distribution and diversity of genus Lilium around the world. In the study, the genetic diversity and genetic relationships of Lilium in China were analyzed by inter-simple sequence Repeat (ISSR) markers. The 6 highly polymorphic ISSR primers were selected to amplify the 20 Lilium species. The results showed that a total of 114 DNA bands were amplified, all of which were polymorphic loci (P = 100%), the effective number of alleles (Ne) was 1.2753, and the difference value between observed number of alleles (Na) and effective number of alleles (Ne) was 0.7247, and the Nei's genetic diversity (H) was 0.2048, and the Shannon's information index (I) was 0.3503. These results indicated that there is significant genetic difference among Lilium species in China. Taking the average genetic similarity coefficient (Gs) 0.5313 as the threshold, the 20 tested Lilium species were clustered into 5 groups, which was not entirely consistent with traditional clustering by morphological traits. The results obtained from this study can provide a reference for the molecular study of Lilium germplasm resources.     

应用微卫星标记分析不同桔小实蝇种群的遗传多样性

为探究不同桔小实蝇Bactrocera dorsalis (Hendel)地理种群的遗传变异、入侵来源和扩散情况,利用13对引物对中国南方10省区、泰国、夏威夷、菲律宾和老挝的30个桔小实蝇种群共180个个体的遗传多样性水平进行了研究。 Popgene32和NTSYS-pc2.10e软件分析结果表明：30个不同桔小实蝇种群的遗传相似度在0.3599~0.9153范围内。种群的Nei氏基因多样性指数平均为0.6464±0.1026,Shannon信息指数I平均为1.2845±0.2632, 提示桔小实蝇种群具有较丰富的遗传多样性。UPGMA聚类分析显示,福建地区和海南地区分别独立一支,广东地区和台湾地区种群聚成一支,而广西、泰国、湖南、云南、老挝、四川、 重庆和贵州地区聚为一大支系。据此提出泰国种群和老挝种群是最早入侵我国的种群,云南地区是最早的入侵地,广西地区可能为又一较早入侵地。     

Deciphering genetic diversity analysis of saffron (Crocus sativus L.) using RAPD and ISSR markers

The existence of genetic diversity in Crocus sativus has globally remained a mystery till date. The study investigated PCR based DNA amplification profile of saffron using ISSR and RAPD based primers. A total of 38 amplicons were generated by ISSR primers in the range from 7 to 12 with an average of 9.50 bands per primer. 20 bands were found to be polymorphic and 18 were monomorphic with an average percentage of polymorphism as 52.48%. RAPD based amplification revealed a total 161 amplicons, 107 as polymorphic and 54 as monomorphic with an average percentage of polymorphism as 66.44%. Cumulative results of RAPD and ISSR demonstrated that Nei-Li’s similarity index ranged between 0.70 and 0.97. The results of AMOVA has revealed 9% of variance among populations and 91% of variance within populations, Φ PT was found as 0.089, which indicates existence of genetic differences though limited. In conclusion, the results indicate that saffron accessions are minimally genetically differentiated, which could be capitalized in future breeding programmes to ameliorate this precious crop.     

广西药用野生稻遗传多样性分析及SSR引物数量对遗传多样性结果的影响研究

采用25对SSR分子标记,对广西境内采集的1610份药用野生稻材料进行遗传多样性和聚类分析,其中检测到等位变异181个,有效等位基因数Ae范围为1.0094(RM240)~2.2674(RM488),平均值为1.3568;期望杂合度He范围为0.0093(RM240)~0.5591(RM448),平均值为0.2112;Shannon多样性指数I在0.0393~0.9296之间变动,平均值为0.3624。数据表明,广西药用野生稻遗传多样性较为丰富,12个药用野生稻地理居群按遗传多样性指数大小排序为:梧州-3南宁-1梧州-2梧州-1南宁-2玉林-2贵港-2梧州-4玉林-3玉林-1贺州贵港-1,其中指数高的居群均分布于梧州市和南宁市两地,因此确定了这两地为广西药用野生稻的遗传多样性中心。通过不同数量的SSR引物对药用野生稻材料间相似系数矩阵进行相关性测验,结果显示当引物按照PIC值降序排列时,10对引物即达到较好聚类效果,升序排列时,21对引物即达到较好聚类效果。研究表明,在进行药用野生稻大居群聚类分析时,SSR引物适宜数量应多于21对,最低不得少于10对。     

Genetic diversity of ten Egyptian chicken strains using 29 microsatellite markers

In this study, we assessed the genetic diversity of three Egyptian local chicken strains (Fayoumi, Dandarawi and Sinai) and six synthetic breeds derived from Fayoumi and Sinai by intercrossing with Barren Plymouth Rock, Rhode Island Red or White Cornish. Diversity measures were based on interrogation of 29 microsatellites. We identified three main clusters of chicken populations encompassing selected Fayoumi lines and Doki-4 (cluster-1), native Dandarawi (cluster-2) and Sinai, and all six synthetic breeds (cluster-3). Dandarawi and Fayoumi lines exhibited lower intra-population genetic diversity and allelic privacy than Sinai and synthetic breeds. The global inbreeding (F(IT) ) was 0.11, among-population differentiation (F(ST) ) was 0.07, and within-population differentiation (F(IS) ) was 0.04. The between-population marker-estimated kinship was lower than within-population estimates. The cluster analysis classified the Fayoumi lines, Dandarawi and Gimmizah as clearly separated populations. The other strains were configured in mosaic admixed groups.