一个芝麻应用核心种质的DNA分子身份证构建

构建并研究应用核心种质是深入挖掘芝麻优异基因和提高育种效率的有效途径。经过对核心种质农艺、形态等性状4年4点的观察统计, 构建了包括大粒、高木酚素、高油等23种应用型在内的131份芝麻应用核心种质。为了准确鉴别这批应用核心种质并对芝麻DNA分子身份证的构建方法进行探索, 本研究利用SSR变性聚丙烯酰胺凝胶电泳技术, 从基于芝麻全基因组开发的32对SSR引物中, 筛选出7对核心引物, 进而采用SSR荧光标记毛细管电泳技术, 共检测出53个多态性位点, 最多一个引物检测到12个多态性位点。将毛细管电泳得到的分子量数据以数字+英文字母方式编码, 用ID Analysis 4.0软件根据最少引物区分最多种质的原则选核心引物中的6对(ZMM1494、ZMM1648、ZMM3037、ZMM2818、ZMM1851、ZMM1935)组合, 构建出如“4A32645(AC017)”这种简单、易使用的字符串DNA分子身份证, 并确定了不同应用型的组内引物组合。还对131份应用核心种质构建了条形码和二维码DNA分子身份证, 可迅速被电子设备识别, 拓宽了DNA分子身份证的使用范围, 并为以后芝麻种质标准化和品种DNA分子身份证库的构建奠定了重要的技术基础。     

东北春播区糜子核心种质及其DNA分子身份证构建

本研究以500份东北春播区糜子资源为材料,利用169个SSR标记,采用UPGMA聚类分组,进行分层抽样,构建核心种质,同时应用ID Analysis 4.0软件构建分子身份证。利用等位基因数(Na)等遗传多样性衡量指标评估核心种质的遗传差异,并利用PCOA分析核心种质。结果表明,对169对SSR引物进行筛选,发现30对多态性好,利用30对SSR引物构建的糜子核心种质包含190份材料,占全部种质的38%,全部种质与核心种质的均检测出91个等位变异,保留了100%等位基因;有效等位基因数为2.2977～2.9975和2.2872～3.0173,平均值分别为2.8198和2.8297;Shannon多样性指数为0.9532～1.0990和0.9535～1.1162,平均值为1.0645和1.0667;观测杂合度为0.3434～0.8037和0.3162～0.7849,平均值为0.5399和0.5359;期望杂合度为0.5654～0.6672和0.5645～0.6707,平均值为0.6448和0.6473;Nei’s基因多样性指数为0.5648～0.6664和0.5628～0.6686,平均值...     

94份谷子核心种质资源分子身份证构建

为科学准确的鉴别谷子种质资源,加强对谷子种质资源管理和新品种保护。筛选出33对SSR标记对94份谷子种质资源进行了分子身份证的构建。从分布在谷子9条染色体上的500多对SSR标记中,筛选出33对多态性标记扩增国内外94份谷子种质资源,检测到203个多态性片段,每对标记的等位基因数为3～10,平均为6.2个。标记位点的多态信息含量(PIC)变幅为0.429～0.864,平均为0.740。品种间特异指数差异较大,为63.336～268.523,平均为189.000。结果表明,根据标记的等位基因数确定10对分子标记b185、b260、b224、b103、b225、CAAS1044、b186、b253、b105、CAAS3008组合,即可将94份核心种质资源完全区分开。     

基于高基元SSR构建黍稷种质资源的分子身份证

为快速鉴定黍稷(Panicum miliaceum)资源,建立大数据管理平台,为种质身份标识和溯源管理提供理论依据。本研究以来源于4个生态栽培区的130份资源为材料,基于35个高基元SSR (四、五和六碱基重复各21、10和4个)构建分子身份证。结果表明, 35个标记中有30个扩增条带稳定,可用于分子身份证构建。30个标记共检出等位变异(Na)90个,平均为30个;有效等位变异(Ne)为2.3186～2.9982,平均为2.7607;Shannon多样性指数(I)为0.9158～1.0873,平均为1.0472;Nei’s基因多样性指数(Nei)为0.5687～0.6665,平均为0.6360;多态性信息含量(PIC)为0.5151～0.7898,平均为0.6966;观测杂合度(Ho)为0.5000～–0.8678,平均为0.7168;期望观测杂合度(He)为0.5710～0.6691,平均为0.6386。基于UPGMA聚类将材料划为3个类群(I、II、III),就山西省材料而言,地方品种和育成品种分别划归类群I和III,农家种在3个类群中均有分布。主成分分析将试材归为4类,聚类结果与...     

基于表型性状和SSR分子标记构建甘薯核心种质

为更好地保存、研究和利用甘薯种质资源,本研究以国家甘薯种质资源圃(广州)保存的1091份甘薯种质为材料,分别采用欧氏距离和Nei’s距离进行NJ聚类分组,组内随机取样,构建核心种质。利用均值、方差、香农多样性指数、变异系数等指标对核心种质的表型性状数据进行代表性评价,以及利用有效等位基因、Nei’s遗传多样性指数、Shannon’s多样性指数等指标对核心种质的SSR分子标记数据进行代表性评价;并利用主成分分析对核心种质进行确认。结果表明,构建的甘薯核心种质包含289份材料,占全部种质的26.49%;在P<0.05概率下,核心种质中表型性状以及SSR分子标记的相关指标与全部种质无显著差异,且二者的表型频率分布基本一致;主成分分析表明核心种质具有与全部种质相似的遗传多样性和群体结构。建立的甘薯核心种质很好地代表了全部种质的遗传变异和群体结构,可为甘薯的品种改良、优良基因挖掘以及种质创新奠定良好基础。     

分子标记及其在云南稻种核心种质中的应用

本文总结了分子生物学发展的各类分子标记及其在核心种质中的应用。     

黄麻应用核心种质的DNA分子身份证构建

It is important to construct and study the applied core collection in order to promote jute genetic breeding and explore excellent genes. In this study, the applied core collection in jute, including 61 accessions divided into 16 application-oriented features, such as high yield, high quality, disease resistance and so on, was established based on the field performance from 300 jute germplasm. 12 fluorescent core primers were screened from 46 pairs of core primers. Fluorescence labeled capillary electrophoresis was used to analyze the polymorphism of these 12 pairs of primers, and a total of 140 polymorphism sites were detected. The precise DNA molecular ID cards were constructed by the combination of the 12 core primer pairs from the data coded in the form of numbers and English letters. Bar code and quick response code DNA molecular were established and can be quickly scanned and recognized by electric gadget. These results could be beneficial to increase the application efficiency and rapid molecular identification in jute germplasm resources.     

DNA分子标记在作物种质资源中的应用进展

近20年来,随着分子生物技术的迅速发展,已发展了多种分子标记,并已得到广泛的应用,文章综述了较为主要的一些分子标记及其在作物种质资源研究领域中的应用.     

杏高通量荧光SSR检测体系及分子身份证的构建

本研究以杏(Prunus armeniaca L.)为试材,根据M13荧光引物和巢式PCR理论,通过调整巢式PCR内两部分循环数、M13荧光引物与正向引物比例和4种荧光(FAM, VIC, NED和PET)产物混合比例3个主要因素,建立了高通量荧光标记SSR检测体系。结果显示,当循环数比设为15∶25、M13荧光引物与正向引物比例为4∶1时,巢式PCR反应体系达到最佳。4种荧光产物按照1μL∶1μL∶1μL∶2μL混合稀释30倍上机检测,所得荧光信号较强、峰高均匀、稳定性好。在建立的体系基础上,筛选了12对多态性良好的李属(Prunus)SSR引物,构建了50份中国杏种质资源的分子身份证。各材料的遗传分子身份证具有唯一性、易读性,可以将种质资源精确的区分开,有利于中国杏种质资源鉴定和保护、品种权益保护和品种创新的激励。     

142份甜高粱品种的分子身份证构建

从分布在高粱染色体10个连锁群上的103对SSR引物中,筛选出41对多态性引物, 并用其扩增国内外142份甜高粱种质资源, 检测到189个多态性片段, 每对引物的等位基因数在2~11之间,平均为4.6个。引物位点的多态信息含量指数(PIC)变幅为0.089~0.850,平均为0.543。品种间特异指数差异较大,介于109.1~454.7之间,平均为189.0。结果表明, 根据引物的等位基因数确定11对引物(Xtxp329、Xtxp258、Xtxp113、Xtxp303、Xtxp61、Xtxp201、Xtxp14、Xtxp91、Xtxp47、Xtxp217和Xtxp67)组合, 并用于构建142份品种资源的分子身份证, 有效地区分了各品种。     

黄麻核心种质的遴选

遴选黄麻核心种质可为黄麻种质创新及新品种选育奠定基础。本研究以300份黄麻种质资源为基础,基于SSR分子标记和农艺性状考察,结合地理来源构建核心种质。结果表明,11个农艺性状变异系数变幅在13.06%～84.87%,表现出丰富的遗传多样性。按农艺性状聚类分析可划分为8个类群,按分子标记聚类可划分为10个类群。结合2个聚类分析、地理位置并按比例取样,建立一个由108份品种(系)组成的预选核心种质。采用44对SSR引物对其进行遗传差异分析,在遗传相似系数为0.65时,可把108份品种(系)分成圆果种和长果种两大类。根据遗传差异分析,剔除遗传相似系数大于或等于0.85的遗传冗余,获得84份品种(系)的核心种质,其中圆果种60份和长果种24份。比较84份核心种质与300份种质的农艺性状变异系数及Shannon-Wiener指数发现,两者之间相差不大,表明遴选的84份核心种质可以最大限度代表300份黄麻种质资源的遗传多样性加以利用和保存。     

黄麻SSR标记与纤维产量性状的相关性

Simple sequence repeat (SSR) markers correlated to fiber yield traits would be a beneficial tool for molecular marker-assisted breeding in jute. In this study, 397 SSR markers were screened from 311 jute germplasms by the phenotype identification from 2016 to 2018 and 116 pairs of primers. The correlation analysis between SSR markers and fiber yield related traits by SPSS revealed that the range of the coefficient of variation related to 9 fiber traits was from 13.05% to 76.78%, indicating a comprehensive genetic variation. The correlation analysis of the agronomic traits showed that there were significantly correlations among these traits. The average correlation coefficient between branch height and nodes of main stem was the highest (r = 0.931), followed by the correlation coefficients (r = 0.781) of fresh bark weight per plant and fresh stem weight per plant and the coefficients (r = 0.779) of dry bark weight and fresh bark weight per plant. Analysis of variance (ANOVA) showed that the traits of main stem nodes, number of branches, plant height and branch height were relatively stable in different years with the higher broad heritability capacity. Furthermore, the SSR markers associated with fiber yield related traits were identified by Pearson correlation method. Stepwise regression analysis among each trait associated SSR markers indicated that there were six markers associated significantly with fiber yield related traits, and phenotypic variation explained by each SSR marker varied from 3.9% to 22.5%. These results will accelerate the development of molecular design breeding in jute.     

基于黄麻转录组序列SNP位点的CAPS标记开发与验证

黄麻CAPS分子标记的开发,可以为黄麻遗传多样性分析、种质资源鉴定和分子标记辅助选择等研究提供有效方法。本试验以黄麻179和爱店野生种为材料,采用IlluminaHiSeq4000测序技术进行转录组测序,对其SNP位点进行分析,设计了与木质素合成基因4CL、COMT及转录因子MYB相兲的SNP引物,在此基础上,应用dCAPS Finder2.0软件开发了CAPS标记,幵对其有效性进行了验证。结果表明:(1)组装后的黄麻unigene序列共72,674条,序列总长度为29,705,997 bp,检测到的SNP位点总数为67,567个,平均每440 bp有1个SNP。(2)获得了39对分别与4CL、COMT和MYB相兲的SNP引物,从中筛选获得26对CAPS标记引物,开发成功率为66.7%,其中11对CAPS标记具有多态性,多态性比例为43.2%。(3)开发的CAPS标记能较好地将12份不同类型的黄麻种质区分开来,表明CAPS是适用于黄麻研究的较理想的分子标记方法,为黄麻遗传基础研究提供了可靠工具。     

上海地区水稻已知品种SSR指纹图谱库构建

用24对引物构建上海地区53份水稻品种的DNA指纹图谱,为当地水稻新品种保护、遗传资源评价及亲缘关系分析提供理论依据和技术支持。结果显示,实验中共检测出109条多态性片段,变幅2~9条,平均每对引物4.5417条多态性片段,有效等位基因数为70.8187,平均每个标记2.9508个;Shannon多样性指数(Ⅰ)平均值为1.1296,变幅0.4949~1.8402;多态性信息含量(PIC)平均值为0.5415,变幅0.2656~0.8001。用24个标记建立53份材料的DNA指纹图谱,这些字符串就构成了这53份材料的“身份证号码”,本实验中,各材料的分子身份证号码是唯一的;聚类分析表明,在遗传相似系数0.21处可以将53份材料分为2个类群,分别为粳稻组和籼稻组,但是组内遗传距离较近。由此可见,上海地区水稻品种的遗传多样性相对狭窄。     

Study of genetic diversity in Indian and exotic sesame (Sesamum indicum L.) germplasm using random amplified polymorphic DNA (RAPD) markers

Fifty-eight accessions of sesame (Sesamum indicum L.), an important oil seed crop of the tropics and subtropics were analysed using random amplified polymorphic DNA (RAPD) technique. The material analysed comprised 36 collections from 18 different states of India and four adjoining countries of the Indian subcontinent, and 22 exotic accessions from 21 sesame growing countries around the world. The results from PCR amplifications with the selected 24 random 10-mer primers were statistically analysed. The value of Jaccard’s similarity coefficients ranged from 0.19 to 0.89. The results indicated the presence of high level of genetic diversity. However, the extent of genetic diversity was greater in the collections from Indian subcontinent as compared to the exotics. Among the Indian accessions, the collections from Rajasthan and North-eastern states were highly diverse. The phenetic analysis grouped 48 out of 58 accessions in six clusters and the remaining highly diverse accessions were placed outside these close-knit clusters. The Bootstrap estimates obtained by Wagner parsimony analysis were significant for seven out of 49 nodes in the majority-rule consensus tree (<95% occurrence). The results of both the analyses were, however, broadly comparable when the constitution of the individual clusters were considered. The principal components analysis indicated that the first two components accounted for only 21% of the total variations and in order to explain <75% of variations 18 components were required. The high level of genetic diversity prevalent among the Indian collections is probably indicative of the nativity of this crop species. Similarly, the relatively lower level of polymorphism in exotic germplasm could be ascribed to the comparatively recent introductions of limited germplasm of this crop into some of the non-traditional sesame growing countries. This revised version was published online in August 2006 with corrections to the Cover Date.     

黄麻纤维产量与主要农艺性状的相关分析

研究黄麻纤维产量与主要农艺性状的相关性, 可为高产育种与生产利用提供科学依据。本文159份不同来源黄麻种质资源的12个农艺性状对纤维产量即单株干皮重的影响表明, 各性状的变异系数变化在11.89%至38.50%之间, 表现出丰富的遗传变异。黄麻纤维产量与各性状均呈极显著正相关, 其中, 与单株鲜皮重、株高、始花期的相关系数较大, 分别为0.814、0.760和0.648。黄麻纤维产量和单株鲜皮重、株高、出麻率、鲜皮厚的回归方程达显著水平, 其标准回归系数依次为0.443、0.437、0.291和0.113。通径分析显示, 单株鲜皮重、株高在决定黄麻纤维产量时起主要作用。出麻率的相关系数(0.253)与直接通径系数(0.291)表现基本一致, 说明出麻率直接对黄麻纤维产量起作用, 具极显著正相关。因此, 在黄麻高产育种中, 应该以始花期、单株鲜皮重、株高、出麻率与鲜皮厚为主要筛选对象, 兼顾综合性状的改良。