基于SCoT分子标记的61份加工型黄瓜种质资源遗传多样性分析及DNA指纹图谱构建

Genetic diversity analysis and DNA fingerprinting construction of 61 processed cucumber germplasm resources based on SCoT markers

摘要

为探明加工型黄瓜种质资源的亲缘关系及遗传多样性，为新品种选育提供理论依据，采用目标起始密码子多态性标记(SCoT)技术，以61份加工型黄瓜种质资源为试验材料，进行遗传多样性分析并构建DNA指纹图谱。结果表明，从100条SCoT引物中筛选出8条扩增条带清晰、多态性高且重复性好的引物；利用筛选出的引物对61份黄瓜种质基因组DNA进行PCR扩增，共扩增出238个位点信息，其中多态性位点227个，平均多态性位点百分比为95.38%；采用NTSYS-pc 2.1软件计算得出供试的61份材料间的遗传相似系数分布在0.588~0.920之间，遗传相似系数最小的材料分别是34号与55号，二者在瓜色、叶片形状及果实形状方面存在较大差异，说明两者亲缘关系最远。基于遗传相似系数对其进行UPGMA聚类分析和树状图绘制，在相似系数0.735处，可将其划分为7个类群，说明61份黄瓜种质材料背景存在差异，但多数地域来源相近、生境相似地区的种质被聚在同一类群。利用2对特异性较大的引物SCoT 12和SCoT 83构建的DNA指纹图谱可单独鉴别出61份黄瓜种质资源，该图谱可为黄瓜分类与鉴定提供科学依据。

Abstract

In order to find out the genetic diversity of germplasm resources of processed cucumber and provide theoretical basis for the breeding of new varieties. the genetic diversity of 61 processed cucumber germplasm resources was analyzed and DNA fingerprinting was constructed using SCoT technique. The results showed that 8 primers with clear amplification bands, high polymorphism and good repeatability were selected from 100 SCoT primers. A total of 238 loci of 61 cucumber germplasm genomic DNA were amplified by PCR using the selected primers, of which 227 were polymorphic loci, and the average percentage of polymorphic loci was 95.38%. NTSYS-pc 2.1 software was used to calculate the distribution of genetic similarity coefficients among the 61 tested materials between 0.588 and 0.920, and the material with the smallest genetic similarity coefficients was 34 and 55, respectively. There were great differences in melon color, leaf shape and fruit shape, indicating that the two were most loosely related. Based on the genetic similarity coefficient, UPGMA cluster analysis and tree diagram were performed. At the similarity coefficient 0.735, they could be divided into 7 groups. The results indicated that 61 cucumber germplasm material backgrounds were different, but most of the germplasm sources were similar, and germplasm from similar habitats were grouped together in the same group. Sixty-one cucumber germplasm resources could be identified by DNA fingerprinting of SCoT 12 and SCoT 83, which could provide scientific basis for classification and identification of cucumber.

关键词

黄瓜 / SCoT分子标记 / 亲缘关系 / 遗传多样性 / DNA指纹图谱

Key words

Cucumber / SCoT molecular marker / Genetic relationship / Genetic diversity / DNA fingerprinting

参考文献

[1] 李亚航, 施艳娥, 丁卓, 等. 黄瓜主要农艺性状分子标记研究进展[J]. 中国瓜菜, 2023, 36(10): 1-9.

[2] 吴仕蔓, 娄兵海, 陈传武, 等. 应用SSR荧光标记法构建22个柚类品种的分子身份证[J]. 果树学报, 2023, 40(4): 605-614.

[3] SAHOO T R, SINGH D K, SINGH N K. Genetic diversity analysis of cucumber (Cucumis sativus L.) genotypes based on ISSR markers[J]. Journal of Pharmacognosy and Phytochemistry, 2020, 9(2): 2131-2136.

[4] MANOHAR S H, HOSAKATTE N M, RAVISHANKAR K V. Genetic diversity in a collection of Cucumis sativus L.assessed by RAPD and ISSR markers[J]. Journal of Plant Biochemistry and Biotechnology, 2012, 22(2): 241-244.

[5] INNARK P, RATANACHAN T, KHANOBDEE C, et al. Downy mildew resistant/susceptible cucumber germplasm (Cucumis sativus L.) genetic diversity assessment using ISSR markers[J]. Crop Protection, 2014, 60: 56-61.

[6] MLIKI A, STAUB J E, ZHANGYONG S, et al. Genetic diversity in african cucumber (Cucumis sativus L.) provides potential for germplasm enhancement[J]. Genetic Resources and Crop Evolution, 2003, 50: 461-468.

[7] 刘亚婷, 罗英, 曾仁杰, 等. 黄瓜种质资源形态学标记和SRAP标记的遗传多样性分析[J]. 中国农学通报, 2017, 33(18): 35-41.

[8] 姚丹青, 楼坚锋, 朱文莹, 等. 基于SNP标记的黄瓜遗传多样性分析[J]. 上海农业学报, 2017, 33(1): 21-30.

[9] 史建磊, 陈先知, 王克磊, 等. 华南型黄瓜种质遗传多样性的SSR分析[J]. 分子植物育种, 2017, 15(6): 2440-2449.

[10] COLLARD B C Y, MACKILL D J. Start codon targeted (SCoT) polymorphism:A simple, novel DNA marker technique for generating gene-targeted markers in plants[J]. Plant Molecular Biology Reporter, 2009, 27(1): 86-93.

[11] 马玉姣, 何西堂, 吴玉森, 等. 基于SCoT分子标记的37份葡萄种质遗传多样性分析[J]. 中外葡萄与葡萄酒, 2023(4): 18-23.

[12] 万雯, 潘鸿, 马涛, 等. 基于SCoT分子标记的广西山核桃种质资源遗传多样性和群体遗传结构分析[J/OL]. 分子植物育种,[2023-03-29]. https://kns.cnki.net/kcms/detail/46.1068.S.20230329.1029.006.html.

[13] 高曼熔, 杨天为, 黄诗宇, 等. 广西野生铁皮石斛SCoT体系优化及遗传多样性分析[J/OL]. 分子植物育种,[2023-08-30]. https://link.cnki.net/urlid/46.1068.s.20230830.0912.002.

[14] 葛聪聪, 王亚娜, 李鸾翔, 等. 香水柠檬×白花柠檬群体子代表型及遗传变异分析[J]. 果树学报, 2022, 39(2): 169-183.

[15] 李欢, 张文洋, 田志强, 等. 高通量分子标记检测方法的研究进展[J]. 玉米科学, 2022, 30(3): 1-9.

[16] 孙蕾倩, 马燕, 李哲, 等. iPBS分子标记研究进展[J]. 分子植物育种, 2022, 20(1): 232-238.

[17] 张珺, 孔维鹏, 刘丽平, 等. ISSR分子标记在果蔬资源上的应用及研究进展[J]. 现代农业科技, 2019(4): 45-47.

[18] 杨梦婷, 黄洲, 干建平, 等. SSR分子标记的研究进展[J]. 杭州师范大学学报(自然科学版), 2019, 18(4): 429-436.

[19] 龙治坚, 范理璋, 徐刚, 等. SCoT分子标记在植物研究中的应用进展[J]. 植物遗传资源学报, 2015, 16(2): 336-343.

[20] 张桂华, 韩毅科, 杨瑞环, 等. 黄瓜种质资源遗传多样性的AFLP分析[J]. 华北农学报, 2007, 22(3): 21-24.

[21] 王惠哲, 李淑菊, 管炜. 应用SRAP标记分析黄瓜的遗传差异[J]. 生物技术通报, 2009(12): 76-79.

[22] 金庆敏, 林毓娥, 吴廷全, 等. 50份黄瓜核心种质的SSR标记筛选与聚类分析[J]. 广东农业科学, 2023, 50(9): 39-48.

[23] 高刚. 国产披碱草属植物的系统与进化研究[D]. 四川雅安: 四川农业大学, 2016.

[24] 王蕾, 王福全, 尹惠萍, 等. 基于SSR标记的黄瓜栽培品种遗传多样性分析及指纹图谱构建[J]. 分子植物育种, 2021, 19(8): 2668-2677.

[25] 卢霞, 刘梦华, 邓志军, 等. 基于InDel标记的黄瓜种质资源遗传多样性分析[J]. 江苏农业科学, 2021, 49(1): 49-54.

[26] 王凤格, 赵久然, 田红丽, 等. 农作物品种DNA指纹库构建研究进展[J]. 分子植物育种, 2015(9): 2118-2126.

基金资助

广西壮族自治区科技先锋队“强农富民”“六个一”专项行动项目(桂农科盟20230505)