摘要
以往进行大白菜杂交种分子纯度鉴定,需要基于亲本开发相应的分子标记,有基因型依赖性。笔者以大白菜杂交种基因组DNA为模板,分别基于自交不亲和S单元型多态性开发的BrCⅠ引物、BrCⅡ引物进行PCR扩增,根据扩增产物的电泳结果确定该大白菜商品杂交种纯度;当杂交种亲本分别为Ⅰ类S单元型和Ⅱ类S单元型时,筛选以BrCⅠ和BrCⅡ引物分别扩增后的产物中只有一个出现条带的材料即为假杂种;当亲本同时为Ⅰ类S单元型或Ⅱ类S单元型时,用限制性内切酶Hinf I或Alu I对PCR产物进行酶切,筛选酶切产物中电泳条带的带型显著不一致的材料为假杂种。该方法为一种新的广适性大白菜杂交种纯度分子鉴定方法,利用该方法进行纯度鉴定,整个周期只需2~3 d,快速精准。
In the past, molecular marker testing of hybrid purity of Chinese cabbage needs to first develop molecular markers based on parents, which are genotype dependent. In this study, the genomic DNA of Chinese cabbage hybrids was used as template, and the amplified products were obtained by PCR amplification with BrCⅠ primer pairs and BrCⅡ primer pairs respectively. The parental type of the commercial hybrids of Chinese cabbage was determined according to the electrophoresis result of the amplified product. When the parents were BrCⅠ S locus and BrCⅡ S locus respectively,only the material with single band was the homozygous parent of the amplification products amplified with BrCⅠ primer pair and BrCⅡ primer pair respectively. When the parents were both BrCⅠ S locus or BrCⅡ S locus, restriction endonuclease Hinf I or Alu I was used for enzyme digestion, and the material with significantly inconsistent electrophoretic bands in the enzyme digestion products was scored as suspected homozygous material. The whole cycle development of this new method only takes 2-3 days, which is fast and accurate.
作者
魏小春
原玉香
赵艳艳
王志勇
杨双娟
苏贺楠
董晓冰
李林
姚春玲
张晓伟
WEI Xiaochun;YUAN Yuxiang;ZHAO Yanyan;WANG Zhiyong;YANG Shuangjuan;SU Henan;DONG Xiaobing;LI Lin;YAO Chunling;ZHANG Xiaowei(Institute of Horticulture,Henan Academy of Agricultural Sciences,Zhengzhou 450002,Henan,China;Yanling Ganluo Health and Old-age Care Co.,Ltd.,yanling 461200,Henan,China)
出处
《中国瓜菜》
CAS
北大核心
2023年第2期19-28,共10页
China Cucurbits And Vegetables
基金
河南省农业科学院杰出青年基金(2021JQ03)
河南省农业科学院自主创新项目(2022ZC21)
河南省重点研发专项(221111110100)
河南省农业良种联合攻关项目(2022010504)
河南省农业科学院科技创新团队(2022TD06)。
关键词
大白菜
S单元型
广适性
分子标记
纯度鉴定
酶切
Chinese cabbage
Slocus
Molecular marker
Purity testing
Restriction enzyme
