猕猴桃性别特异性毛细管荧光分子标记开发  

Development of specific markers for the characterization of kiwifruit sex trait based on fluorescence capillary electrophoresis

作　　者：方林川 张鸿 陈志伟 王发明[2] 戢小梅 ANG Linchuan;ZHANG Hong;CHEN Zhiwei;WANG Faming;JI Xiaomei(Wuhan Academy of Agricultural Sciences,Wuhan,Hubei,430074,China;Guangxi Institute of Botany,Chinese Academy of Sciences,Guilin,Guangxi,541006,China)

机构地区：[1]武汉市农业科学院,武汉430074 [2]广西壮族自治区中国科学院广西植物研究所,桂林541006

出　　处：《中国南方果树》2024年第3期7-15,共9页South China Fruits

基　　金：湖北省重点研发计划(2021BBA090,湖北低海拔地区猕猴桃适宜品种筛选及配套优质高效栽培技术研究与示范);湖北省重点研发计划(2023BBB064,猕猴桃集约化高效育苗技术创新及应用);2021年武汉市猕猴桃发展专项(武汉市猕猴桃种苗健康繁育关键技术研发与应用)资助。

摘　　要：猕猴桃是起源于我国的雌雄异株果树。通过开发与猕猴桃性别表型关联的功能标记,可在苗期准确判定杂交后代或种质资源的性别,极大提高猕猴桃早期选择的效率,从而为种质资源创新奠定基础。基于猕猴桃Y染色体性别决定非重组区中抑雌基因SyGl与促雄基因FrBy的外显子保守序列设计引物,利用不同群体材料对其进行有效性验证,从中筛选出高效引物对,利用琼脂糖凝胶电泳、目的片段重测序与荧光毛细管电泳方法比较了不同分子标记的效果,通过引入内参ITS引物,建立了多重PCR体系及荧光毛细管电泳检测体系,利用389份样品对检测体系的效果进行检验。结果表明:筛选获得2个性别特异性标记(Ms1及Ms2),可用于猕猴桃性别鉴定。内参ITS引物的引入,降低了DNA质量造成的实验误差。开发出的多重(ITS、Ms1及Ms2)PCR体系及荧光毛细管电泳检测体系,对包括中华猕猴桃品种、美味猕猴桃品种、野生资源(对萼猕猴桃、毛花猕猴桃、华南猕猴桃、奶果猕猴桃)和中华猕猴桃杂交后代材料在内的389份样品进行检测,有370份样品的性别被准确鉴定,总体鉴定准确性达到95.11%。该检测体系,可提高性别标记的检测效率,适用于猕猴桃性别性状的分子辅助与设计育种。Kiwifruit is a dioecious fruit that originated in China.By developing functional markers associated with the sex determination of kiwifruit,the sex of hybrid offspring or germplasm resources at the seedling stage could be detected,which greatly improved the efficiency of early sex trait selection,thus laying a foundation for germplasm resource innovation in an economical and efficient manner.Primers were designed based on the conserved exon sequences of female suppress gene SyGl and male improve gene FrBy in the sex determination non-recombinant region of Y chromosome of kiwifruit,the effectiveness was proven by different population materials,and the high efficiency primer pair was screened.Effect of different molecular markers was compared by using agarose gel electrophoresis,target fragment sequencing and fluorescence capillary electrophoresis methods.By introducing internal reference ITS primers,multiple PCR system and fluorescence capillary electrophoresis detection system were established,and the effect of the detection system was tested by 389 samples.The results showed that two sex-specific markers were obtained including Ms1 and Ms2,which could be used for sex identification of kiwifruit.Introduction of internal reference ITS primer decreased the experiment error caused by DNA quality.Three hundred and eighty-nine samples including varieties of Actinidia chinensis Planch.,A.deliciosa and wild germplasm resources such as A.valvata,A.eriantha,A.glaucophylla and A.carnosifolia var.glaucescens were tested by multiple PCR system(ITS,Ms1,Ms2)and fluorescence capillary electrophoresis detection system.Sex of 370 samples was accurately identified,with accuracy rate of 95.11%.The sex detection system developed in this study could improve the identification efficiency of sex marker,is suitable for molecular marker assisted selection of sex traits in kiwifruit.

关 键 词：猕猴桃 性别鉴定 种质资源 分子标记辅助选择 

分 类 号：S66[农业科学—果树学]