一个芝麻应用核心种质的DNA分子身份证构建
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摘要
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。
The establishment and research of sesame applied core collection are significantly effective to facilitate the improvement of breeding techniques and exploration of out-standing genes. A sesame applied core collection, including 131 sesame accessions with 23 application-oriented features such as big seeds, high oil content and high lignan content, was established according to the observation and statistics in four various environments for four years. Based on 32 pairs of SSR primers developed from the sesame genome, we screened out seven pairs of primers, bearing high polymorphism, distinct bands and excellent repetition, using the technique of denatured polyacrylamide gel electrophoresis for detecting SSR The more precise detection technique of capillary electrophoresis with fluorescent SSR markers was utilized simultaneously to analyze the 131 sesame germplasm of applied core collection. Totally 53 polymorphic locations were detected out by using six primer pairs, and the most sensitive primer pair could detect 12 polymorphic locations. Subsequently, the results were shown in the form of numbers combined with English letters. As per the rule that the least primer pairs detect the most sesame germplasm, the combinations of six core primer pairs, namely ZMM1494, ZMM1648, ZMM3037, ZMM2818, ZMM1851, and ZMM1935 effectively distinguishing the 131 sesame germplasm of applied core collection were screened out by ID Analysis 4.0. So that, the simple and easy-applied DNA mo-lecular identifications such as "4 A32645(AC017)" were constructed, and the various combinations of primers distinguishing the germplasm within groups were screened out simultaneously. The DNA molecular identifications of character strings, bar code and quick response(QR) codes, easily scanned and recognized by electric gadget, were constructed based on the molecular data of 131 sesame germplasm of applied core collection so as to broaden their application ranges, also provide important technical foundation for the standardization of sesame germplasm and the construction of DNA identification bank of sesame varieties.
The establishment and research of sesame applied core collection are significantly effective to facilitate the improvement of breeding techniques and exploration of out-standing genes. A sesame applied core collection, including 131 sesame accessions with 23 application-oriented features such as big seeds, high oil content and high lignan content, was established according to the observation and statistics in four various environments for four years. Based on 32 pairs of SSR primers developed from the sesame genome, we screened out seven pairs of primers, bearing high polymorphism, distinct bands and excellent repetition, using the technique of denatured polyacrylamide gel electrophoresis for detecting SSR The more precise detection technique of capillary electrophoresis with fluorescent SSR markers was utilized simultaneously to analyze the 131 sesame germplasm of applied core collection. Totally 53 polymorphic locations were detected out by using six primer pairs, and the most sensitive primer pair could detect 12 polymorphic locations. Subsequently, the results were shown in the form of numbers combined with English letters. As per the rule that the least primer pairs detect the most sesame germplasm, the combinations of six core primer pairs, namely ZMM1494, ZMM1648, ZMM3037, ZMM2818, ZMM1851, and ZMM1935 effectively distinguishing the 131 sesame germplasm of applied core collection were screened out by ID Analysis 4.0. So that, the simple and easy-applied DNA mo-lecular identifications such as "4 A32645(AC017)" were constructed, and the various combinations of primers distinguishing the germplasm within groups were screened out simultaneously. The DNA molecular identifications of character strings, bar code and quick response(QR) codes, easily scanned and recognized by electric gadget, were constructed based on the molecular data of 131 sesame germplasm of applied core collection so as to broaden their application ranges, also provide important technical foundation for the standardization of sesame germplasm and the construction of DNA identification bank of sesame varieties.
引文
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