内蒙古糜黍资源DNA分子身份证的构建
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1.山西农业大学;2.山西省农业科学院农作物品种资源研究所;3.内布拉斯加大学林肯分校农艺系小宗粮豆研究与推广中心

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国家现代农业产业技术体系建设专项(CARS-06-14.5-A16);山西省现代农业产业技术体系建设专项资金项目(2023CYJSTX03-12);山西省重点研发项目(2022ZDYF110)


Construction of DNA Molecular Identity Card of Broomcorn Millet Resources in Inner Mongolia
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the National Special Fund for Construction of Modern Agricultural Industrial Technology System (CARS-06-14.5-A16); the Special Fund for Construction of Modern Agricultural Industrial Technology System of Shanxi Province (2023CYJSTX03-12); the Key Research & Development Project of Shanxi Province (2022ZDYF110)

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    摘要:

    糜黍(Panicum miliaceum L.)是一种品种资源多样的古老作物,采用荧光SSR标记,对糜黍资源进行数字化的管理与分类,可将众多资源合理高效应用。本研究以258份内蒙古糜黍资源为材料,基于本课题组前期开发的85个常规SSR标记,经多次扩增筛选、优化、加工(引物5′端添加荧光基团)获得荧光SSR核心标记。利用MapChart 2.32绘制标记的染色体定位图,利用ID Analysis 4.0检测标记对材料的区分度,用PowerMarker 3.25和PopGen 1.32进行遗传多样性分析,用 MEGA 11.0.10和NTSYSpc2.11a分别绘制聚类图和主成分分析图,最终应用草料二维码生成器构建材料的DNA分子身份证。遗传多样性分析发现,利用7个标记扩增258份材料,共有123个等位基因(Na),平均每个标记有17.5714个,处于8.0000~25.0000之间;有效等位基因数(Ne)变化范围在3.5868~16.2042之间,均值是7.4622;Shannon多样性指数(I)处于1.8361~2.8852之间,平均值为2.2270;观测杂合度(Ho)、期望观测杂合度(He)分别处于0.5303~0.9606、0.7229~0.9408之间,均值分别为0.8021和0.8372;Nei’s基因多样性指数(Nei)变化范围在0.7212~0.9383之间,均值是0.8353;多态性信息含量(PIC)为0.8266~0.9391,均值是0.8994。将毛细管电泳结果以特定方式进行编码处理,最终仅用7个荧光标记(RYW6、RYW8、RYW37、RYW40、RYW67、RYW124和RYW125)组合在一起生成全部材料的字符串和二维码DNA分子身份证,为内蒙古糜黍种质资源的分类管理及快速鉴定提供了分子检测工具和理论依据。

    Abstract:

    Broomcorn millet (Panicum miliaceum L.) is an ancient crop with various varieties and resources. Using fluorescent SSR markers, the resources of Panicum miliaceum L. can be managed and classified digitally, and many resources can be applied rationally and efficiently. In this study, 258 samples of millet resources from Inner Mongolia were used as experimental materials. Based on 85 pairs of conventional SSR markers developed by our research group, fluorescent SSR core markers were obtained through multiple amplification, screening, optimization and processing (fluorophores were added at the 5 'end of the primer). MapChart 2.32 was used to draw the chromosome location map of core markers, ID Analysis 4.0 was used to detect the differentiation of markers to materials, and PowerMarker 3.25 and PopGen 1.32 were used to analyze genetic diversity. MEGA 11.0.10 was used to draw the cluster diagram, NTSYSpc2.11a was used to draw the principal component analysis diagram, and finally the grass QR code generator was used to build DNA molecular identity cards of all materials. Genetic diversity analysis showed that there were 123 alleles (Na) in 258 materials amplified by 7 pairs of markers, with an average of 17.5714 NA per pair, ranging from 8.0000 to 25.0000. The number of effective alleles (Ne) ranged from 3.5868 to 16.2042, with an average of 7.4622. Shannon diversity index (I) was between 1.8361 and 2.8852, with an average value of 2.2270. Observed heterozygosity (Ho) and expected heterozygosity (He) ranged from 0.5303 to 0.9606 and 0.7229 to 0.9408, respectively, and the mean values were 0.8021 and 0.8372. Nei's gene diversity index (Nei) ranged from 0.7212 to 0.9383, with an average of 0.8353. Polymorphism information content (PIC) ranged from 0.8266 to 0.9391, with an average of 0.8994. The results of capillary electrophoresis were encoded in a specific way, and only 7 pairs of fluorescent labels (RYW6, RYW8, RYW37, RYW40, RYW67, RYW124 and RYW125) were used to generate 258 strings and two-dimensional code DNA molecular identity cards of Inner Mongolia millet resources. The results provided a reference for the classification management and rapid identification of the germplasm resources of Inner Mongolia broomcorn millet.

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  • 收稿日期:2024-04-03
  • 最后修改日期:2024-05-03
  • 录用日期:2024-09-10
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