中国主要棉花栽培群体的遗传多样性分析

利用ISSR分子标记对我国棉花栽培品种中的38个陆地棉(Gossypium hirsutum)和6个海岛棉(Gossypium bar-badense)共44个品种(按来源划分为6个品种群,6个海岛棉品种均属于新疆品种群体)进行遗传多样性分析表明,筛选出的11个引物在所有品种中共扩增出90条带,其中70条为多态性条带,多态性条带百分率(PPB)达77.78%,Popgene32软件分析结果表明,新疆品种群的遗传多样性水平最高[PPB=91.43%,H(基因多样性指数)=0.3769,I(信息指数)=0.5464],其次是中棉所品种群,河南品种群排位第三,而河北、山东和山西3个品种群体的遗传多样性水平相差较小,其中山西品种群遗传多样性水平最低(PPB=58.57%,H=0.2484,J=0.3587).6个地方棉花栽培群体在总的遗传变异中有82.42%存在于群体内,群体间的遗传变异仅占总变异的17.58%,群体间分化系数Gst=0.1759,群体间总的基因流的估算值.Nn=2.34.聚类分析结果表明,山西、山东和河北品种群体的亲缘关系最近,先聚合在一起,然后依次与河南和中棉所品种群体聚类,最后与亲缘关系最远的新疆品种群体聚合.     

我国主要棉花栽培群体的遗传多样性分析

摘 要：利用ISSR分子标记对我国44个棉花栽培品种（拟似为6个群体）进行遗传多样性分析。筛选出的11个引物在所有品种中共扩增出90条带,其中70条为多态性条带,多态性条带百分率(PPB)达76.1%,Popgene32软件分析结果表明,新疆种群的遗传多样性水平最高(PPL=91.43%,H=0.3769,I=0.5464),其次是中棉种群、河南种群,而河北、山东、山西3个种群的遗传多样性水平相差较小,6个棉花种群在其总的遗传变异中有82.42%的存在于群体内,群体间的遗传变异仅占总变异的17.58%,群体分化系数Gst=0.1759,群体间总的基因流的估算值Nm=2.34。由聚类分析可知,山西、山东、河北种群的亲缘关系最近,先聚合在一起,然后依次与河南、中棉所种群聚类,最后与亲缘关系最远的新疆种群聚合。     

采用Nested -PCR从田间和水稻植株上检测稻曲病菌

采用稻曲病菌(Ustilaginoidea virens)的特异性引物，对从发病程度不同的稻田土样、水稻（Oryza sativa）生殖生长初期稻田浮萍(Lemma ssp.)和水稻植株的DNA进行Nested PCR扩增，分析稻曲病菌在稻田的存在和在植株上的附着和侵染，结果显示在稻田中稻曲病菌可以附着在浮萍上，从第一年种植水稻田块的土壤中未检测到稻曲病菌，但在稻田中的浮萍上可以检测到；水稻生殖生长初期剑叶的叶耳间已有稻曲病菌附着和侵染。实验证明PCR技术可以有效地检测田间稻曲病菌。关键词：稻曲病菌(Usitilaginoidea virens); 检测; Nested PCR     

利用SSR和RAPD标记分析中国部分地区晚疫病菌群体遗传多样性

利用SSR和RAPD对来自福建、黑龙江、河北和内蒙古4个地理群体的80个马铃薯晚疫病菌（phytophthora infestans）株进行了遗传多样性分析。13对SSR引物共扩增出76条谱带,多态性条带比率78.9%,相似系数变化范围0.00～0.42之间;筛选出的14条RAPD引物共扩增出189条谱带,多态性条带比率95.2%,相似系数变化范围0.04～0.66之间。遗传多样性分析表明,在4个群体中,福建群体的多样性更为丰富。遗传相似性分析显示,黑龙江和内蒙古两个群体间的遗传相似性最高,而福建和河北两个群体间的遗传相似性最低。聚类分析显示,来自南方福建的菌株与来自北方黑龙江、河北和内蒙古的菌株亲缘关系较远,且福建群体分布于更多的聚类组,显示出更高的遗传变异度。     

绿色豌豆蚜不同地理种群的遗传多样性

豌豆蚜是我国苜蓿上危害最为严重的害虫之一。利用微卫星标记研究了我国绿色型豌豆蚜10个地理种群的遗传相似性、基因分化、遗传距离与地理距离和海拔之间的关系及其基因结构。选取15对引物扩增300个个体,共检测到41个多态性条带,多态性条带百分率(PPB)为100%。10个豌豆蚜地理种群观测等位基因数(Na)为1.592 7,有效等位基因数(Ne)为1.356 9,Nei’s基因多样性指数为0.206 6,Shannon-Wiener指数(I)为0.307 6。新疆维吾尔自治区、陕西省、河南省种群的遗传多样性较高,内蒙古自治区、山东省、青海省种群相对较低。然而,10个地理种群豌豆蚜(绿色型)的遗传分化较高(Gst=0.399 6)。种群聚类分析结果显示,全部豌豆蚜种群明显聚为两大类群,山东省、河南省种群为一大类群,其余为另一大类群。Mentel检测表明,遗传分化与地理距离、海拔无显著相关性。我国豌豆蚜种群具有非常丰富的遗传多样性,应加强豌豆蚜的监测和治理。     

应用RAPD技术研究4种鲍的亲缘关系

应用RAPD技术研究 4种鲍的亲缘关系结果表明 ,4种鲍群体中 2 0个有效引物共扩增出 5 38条DNA带 ,平均每个引物扩增的条带数为 2 6 .9;扩增的多态性条带数 136 ,平均每个引物扩增的多态性条带数为 6 .8;多态位点百分数为 2 5 .3%。盘鲍群体和皱纹盘鲍群体之间遗传距离与遗传一致度分别为 0 .2 8和 0 .72 ,杂色鲍群体和九孔鲍群体之间遗传距离与遗传一致度分别为 0 .32和 0 .6 8。聚类分析把盘鲍群体和皱纹盘鲍群体聚为 1组 ,二者亲缘关系较近 ;杂色鲍群体和九孔鲍群体聚为 1组 ,二者亲缘关系也较近     

青钱柳种质资源多样性SRAP初步分析

为了利用分子标记方法评价青钱柳种质资源的遗传多样性,本文对青钱柳DNA的提取、SRAP扩增体系重要参数进行了优化,运用优化体系筛选多态性引物,并用1对引物对青钱柳9个种源进行了遗传多样性初步分析。研究结果建立了适于青钱柳SRAP的扩增体系;从110对SRAP引物中筛选出了13对多态性引物,运用1对引物组合Me7＋Em2扩增获得21个多态性位点,多态率达100%。遗传多样性分析表明有效等位基因数（Ne）为1.3429,平均Shannon＇s信息指数（I）为0.3687,Nei＇s基因多样性指数（H）为0.2267,种源的遗传分化指数Gst为0.1983;聚类分析结果表明9个种源在遗传距离0.100处聚为3类,聚类结果和地理距离之间呈现较高的相关性。本文的研究结果表明所建立的青钱柳种质资源库具有广泛的遗传多样性,为进一步的开发利用提供了条件。     

47份水稻品种资源的ISSR遗传多样性分析

为研究广东省惠州市种植的常规水稻品种的遗传多样性,本实验利用ISSR标记对47份水稻品种资源进行遗传多样性检测。从49条引物中筛选出5条重复性好,条带清晰的引物进行PCR扩增,共扩增出53条带,每个引物可以扩增出9～13条带,平均为10．6条,其中47条具有多态性,比率为88．7％。不同水稻品种间遗传相似系数变幅为0．319～0．936,平均达0．691,说明ISSR标记能够揭示材料间较高的遗传多样性。通过聚类,从分子水平对水稻品种资源的遗传关系进行分析,并对47份水稻品种资源进行分类,ISSR标记能将47份水稻品种完全区分开,为水稻品种资源的研究利用提供参考。     

RAPD和ISSR标记对甜瓜种质遗传多样性的研究

利用RAPD和ISSR两种分子标记技术对37份甜瓜(CucumismeloL.)种质进行了遗传多样性研究.在供试材料中筛选到具有多态性的RAPD引物21个,ISSR引物10个(其中ISSR-2与ISSR-4等量混合组成ISSR-10引物对).RAPD引物共扩增出多态性带106条,多态性条带比率(PPB)为58.62%,平均多态信息量(PIC)为0.47;ISSR引物共扩增出多态性带73条,PPB值为65.51%,平均PIC值为0.53.根据两种标记的结果,采用UPGMA聚类分析,将供试材料分为两大类群野生甜瓜和栽培甜瓜;栽培甜瓜又分为厚皮甜瓜和薄皮甜瓜两大亚群,各野生甜瓜种质之间的遗传距离较大,这与其分类地位基本一致.两种分子标记的分析结果呈极显著正相关(r=0.62>r=0.01).研究表明,RAPD和ISSR标记可用于甜瓜种质遗传多样性的研究.     

紫茎泽兰群体遗传多样性及遗传结构的AFLP分析

紫茎泽兰是我国最为严重的入侵物种之一。本研究以中国境内的紫茎泽兰为材料，用AFLP方法研究了其群体遗传多样性及遗传分化。用筛选出的3对荧光引物，对5个地区62个群体进行AFLP分析，共扩增出490条带，其中多态性带328条，群体的遗传多样性较高(P = 66.9%，H = 0.171，I = 0.268)，主要存在于群体内，群体间的遗传分化系数Fst为0.287。AMOVA分析表明，在地区水平紫茎泽兰的遗传分化主要存地区内，群体内遗传分化占 70.25%，群体间遗传分化占21.71%，地区间的遗传分化仅占8.04%。UPGMA聚类分析结果表明，62个地理群体主要分为四大类群，并且有明显的地缘关系。Sigmaplot分析表明海拔是影响紫茎泽兰遗传多样性主要地理因子。Mantel检验表明遗传距离与地理距离成正相关(r = 0.34)，但也有例外。由此推断风媒传播可能是紫茎泽兰的主要传播方式，水媒传播可能是紫茎泽兰的另一主要传播途径；随着紫茎泽向东、向北扩散，新入侵地区的遗传多样性逐步降低。     

8份广西产绞股蓝种质的RAPD引物筛选及其遗传多样性分析

本研究的目的在于筛选合适的RAPD随机引物,应用RAPD技术对药用植物绞股蓝进行遗传多样性分析,并构建DNA指纹图谱。研究结果表明,我们利用生物信息学方法挑选出的20条引物中有19条引物的扩增条带清晰且多态性好;在清晰稳定出现的354条带中,294条具有多态性;其中有3条引物的扩增条带可清楚区分绞股蓝与混淆品种乌蔹莓,可建立其DNA指纹图谱。按UPGMA法进行聚类分析,计算其遗传相似系数,结果显示,8份绞股蓝供试材料聚为两类,聚类结果与其地理区域远近和生长环境一致。本研究中筛选出的19条引物适用于绞股蓝遗传多样性分析,且获得的DNA指纹图谱可用于鉴别绞股蓝。     

尼里——拉菲奶水牛的RAPD分析

以尼里—拉菲奶水牛基因池为模板,从22条10 bp的随机引物中筛选出17条具有清晰多态条带的引物,从中选取5条,对尼里—拉菲奶水牛基因组DNA进行RAPD扩增,结果得到53条带,其中12条表现多态,多态频率23.85%;根据N e i氏公式计算品种内的遗传相似指数F的平均值为0.9500,遗传距离指数D的平均值为0.0500。     

Analysis of genetic diversity of soybean germplasm from five different origins using RAPD markers

The pattern of genetic diversity among 92 genotypes of soybean from 5 different origins/sources (Pakistan, the USA, Asian Vegetable Research Development Centre (AVRDC), Japan and North Korea) was analyzed using randomly amplified polymorphic DNA markers. Out of 20 random primers 6 tested, 10 were polymorphic among genotypes and they yielded 107 markers, with an average of 10.7 markers per primer. The proportion of polymorphic bands within genotypes ranged from 0.47 to 0.71 with an average of 0.59. Pakistani and US genotypes exhibited the highest number of polymorphic bands (95%), while North Korean genotypes revealed the lowest (60%). The mean band frequency of the primers among genotypes was 0.57 with a range of 0.08–0.99. The Shannon’s index and Nei’s genetic diversity index revealed that primer OPF-06 showed maximum genetic diversity among the genotypes. Dendrogram constructed using Unweighted Pair Group Mean Average (UPGMA) method divided the genotypes into 5 main groups consisting of 13 clusters. The results of cluster analysis indicated that the genetic diversity between Pakistani and US or AVRDC genotypes is much larger than that between Pakistani and North Korean or Japanese genotypes. The Pakistani genotypes had distinct bands from plant introductions. Therefore, the Pakistani genotypes may be useful to soybean breeders.     

Abundant genetic diversity in cultivated <Emphasis Type="Italic">Codonopsis pilosula</Emphasis> populations revealed by RAPD polymorphisms

Genetic diversity of seven cultivated populations of Codonopsis pilosula Nannf. from Longxi County, Gansu Province of China was estimated using randomly amplified polymorphic DNA (RAPD) markers. The 17 selected RAPD primers amplified 205 polymorphic bands out of a total of 235 (87.2%). Nei’s gene-diversity statistics and population differentiation parameters based on AMOVA analysis indicated that the cultivated C. pilosula populations remained a high level of genetic diversity with Hs = 0.299 and I = 0.450. A greater proportion of genetic diversity was found within (77%) rather than among (23%) the populations. In addition, we also detected that populations from different altitudes had a considerable genetic differentiation after 40 years of cultivation at the same site. Populations from higher altitude had lower genetic diversity than those from lower altitude. Our results suggested that irregular and sparse cultivation practices, i.e., random collecting, preserving, and planting seeds of the medicinal species without deliberate selection, might be an efficient way to conserve genetic resources of medicinal plants, in addition to their effective uses.     

Genetic diversity and relationships among safflower (<Emphasis Type="Italic">Carthamus tinctorius</Emphasis> L.) analyzed by inter-simple sequence repeats (ISSRs)

Genetic diversity and relationships among 48 safflower accessions were evaluated using 22 inter-simple sequence repeats (ISSR) primers. A total of 429 bands were amplified, and 355 bands (about 82.7%) were polymorphic. Five to forty-one polymorphic bands could be amplified by each primer, with an average of 16.1 polymorphic bands per primer. The results showed that the polymorphism of the safflower germplasm was higher at the DNA level. All the 48 accessions could be distinguished by ISSR markers and were divided into 9 groups based on ISSR GS by using UPGMA method. The genetic relationships among the accessions from different continents were closer. Comparatively, the genetic diversity of the accessions originated from Asia was higher, from Europe assembled. The results also showed that the genetic variation of accessions from Indian and Middle Eastern safflower diversity centers were relatively higher. ISSR is an effective and promising marker system for detecting genetic diversity among safflower and give some useful information on its phylogenic relationships.     

Genetic diversity of Pakistan wheat germplasm as revealed by RAPD markers

Wheat breeding in Pakistan started in 1930s before partition in the United India and so far has released more than 68 cultivars, but no systematic analyses of the genetic diversity of Pakistan wheat have been made. Twenty Pakistan wheat cultivars released from 1933 to 2002 were examined for genetic diversity and relationships using random amplified polymorphic DNA (RAPD) markers. Forty-two RAPD primers were applied and 184 polymorphic bands were generated for each cultivar. Most of the cultivars were genetically interrelated, although six of them displayed some genetic distinctness. The RAPD variation observed among these cultivars was low. Only 40.7% of the total scorable bands were polymorphic, and 26.1% of the polymorphic bands were observed most frequently (f = 0.95) among the 20 cultivars. The proportions of polymorphic bands for each cultivar ranged from 0.67 in ‘Yecora’ to 0.84 in ‘C-250’ with an average of 0.76. About 1.4% of the RAPD variation might have been fixed over the 69 years of wheat breeding, but such fixation was not statistically significant. These results are significant for future improvement and conservation of Pakistan wheat.     

Genetic diversity analysis of hulless barley from Shangri-la region revealed by SSR and AFLP markers

For adding the hulless barley resources of Shangri-la region to the global barley resource library, a basic work was done by us to assess their genetic diversity of this region. The genetic diversity of 60 hulless barley samples collected from three counties in Shangri-la region of Yunnan Province, were studied using SSR (simple sequence repeats) and AFLP (amplified fragment length polymorphism) markers. A total of 70 alleles were detected for 19 pairs of SSR primers, and 525 band containing 464 polymorphic bands were revealed for 5 pairs of AFLP primers. The value of polymorphism information content (PIC) ranged from 0.03 to 0.86 for SSR primers. The total numbers of alleles were 51, 55, 43 in three populations and the polymorphic bands were 188, 205 and 141. The genetic distances and genetic identity among the three populations showed their close relationship. The gene diversity among populations relative to the total population diversity (Gst) was 0.13 for SSR markers and 0.02 for AFLP markers and indicated that just 13 and 2% variations were among populations, respectively. The UPGMA cluster analysis revealed that all of the samples grouped randomly rather than clustered into distinct groups corresponding to their populations, row types and spring/fall types. We concluded that there was high genetic diversity in the population of Shangri-la region and the formation of diversity was related to complex environment and inhabitants’ traditional practices.     

草莓品种的亲缘关系及其不同继代次数的叶片组培苗的RAPD分析

从100个随机引物中筛选出10个引物,对16个草莓品种进行RAPD分析。结果显示,10个引物共扩增出73条DNA谱带,其中多态条带49条,多态性程度为67.13%;遗传距离D值在0.40水平上能将供试材料聚为3类,表明RAPD能很好地鉴别草莓品种之间的遗传关系。利用前面筛选的10个随机引物和其他20个随机引物对不同继代次数的草莓组培继代苗进行了RAPD分析,其特征带型表现一致,未发现变异,说明草莓叶盘离体继代培养30次以内能稳定遗传。