不同遗传背景腐烂茎线虫生物学杂交体系构建与验证  

Construction and Verification of Biological Hybridization System of Different Genetic Background Ditylenchus destructor

作　　者：王宏宝[1,2] 毛佳 曹凯歌[1] 付佑胜 吴险平[1] Wang Hongbao;Mao Jia;Cao Kaige;Fu Yousheng;Wu Xianping(Huaiyin Institute of Agricultural Sciences of Xuhuai Region in Jiangsu,Huai'an 223001,Jiangsu;Jiangsu Key Laboratory for Eco-Agricultural Biotechnology around Hongze Lake(Huaiyin Normal University),Huai'an 223300,Jiangsu;Huai'an Agricultural Science and Technology Industry Corporation,Huaian 223001,Jiangsu)

机构地区：[1]江苏徐淮地区淮阴农业科学研究所,江苏淮安223001 [2]江苏省环洪泽湖生态农业生物技术重点实验室(淮阴师范学院),江苏淮安223300 [3]江苏省淮安市农业科技实业总公司,江苏淮安223001

出　　处：《长江大学学报（自然科学版）》2019年第6期56-59,I0004,共5页Journal of Yangtze University(Natural Science Edition)

基　　金：江苏省环洪泽湖生态农业生物技术重点实验室项目(17HZHL016);江苏省农业自主创新项目(CX(14)5086);江苏省淮安市农科院科研发展基金项目(201906)

摘　　要：为研究不同群体腐烂茎线虫(Ditylenchus destructor)生物学杂交体系的构建方案,分别通过薯片平板杂交法、薯条离心管杂交法、薯块切片打孔杂交法构建3种生物学杂交体系探讨了不同遗传背景线虫群体间杂交产生后代的情况。结果显示,3种杂交构建方案均可以培养成功,不同的是甘薯切片接种法杂交率高、后代数量多,亲本自交数量与其他2种方法相比差异显著,同时甘薯切片法微环境更接近原始寄主状态,操作简便、省时省力,相比甘薯平板打孔法和薯条离心管杂交法更便于线虫在小孔内集中,增加线虫杂交几率。比较不同杂交体系优缺点,认为甘薯切片打孔法所构体系为腐烂茎线虫的生物学杂交试验最佳体系。同时,试验对“DeLY×DeYL”杂交后代进行分子鉴定,验证杂交体系是成功的。该体系的建立可揭示不同地理来源的具有不同遗传背景的腐烂茎线虫群体间杂交在后代遗传背景上表现异质性。该结果可为今后不同遗传背景线虫间进行生物学杂交提供技术指导,并为不同遗传背景线虫的生殖隔离与其他进境检疫性线虫的生物学杂交研究提供参考。To study the construction of biological hybridization system of different genotypes of stem nematode( Ditylenchus destructor ),three biological hybridization systems,such as potato chip flat hybridization,French fries centrifuge tube hybridization and potato chip slice punching hybridization,are constructed to explore whether different genotypes of nematode populations can be crossed to produce offspring.The results show that three hybridization schemes can be successfully cultured.The difference is that the hybridization rate of sweet potato slice inoculation method is high,and the number of offspring is large.The number of parental self-crossing is significantly different from the other two methods.The sweet potato slice micro-environment is closer to the original host state,and the operation is simple and it can save time and effort.Compared with the sweet potato plate punching method and the French fries centrifuge tube hybrid method,it is more convenient for the nematodes to concentrate in the small holes and increases the probability of nematode hybridization.Compared the advantages and disadvantages of different hybridization systems,the biological hybridization test of the rotted stem nematode is carried out by using the sweet potato slice punching method as the best system.Molecular identification of the offspring of “DeLY×DeYL” hybrids is carried out.The results show that the hybrid system is successful.At the same time,it also reveals that the hybridization between populations of rotten stem nematodes from different geographical sources and with different genetic backgrounds show heterogeneity in the genotypes of their offspring.The results provide a technical guidance for biological hybridization between different genotypes of nematodes in the future,and it also provides reference for reproductive isolation of different genotypes of nematodes and biological hybridization of other imported quarantine nematodes.

关 键 词：腐烂茎线虫(Ditylenchus destructor) 生物学杂交 分子鉴定 

分 类 号：S432.45[农业科学—植物病理学]