摘要
为了实现黏类小麦雄性不育基因rfv1的定向转育,创制优良的黏类非1BL/1RS小麦雄性不育新保持系,本研究以1BS上带有不育基因rfv1的非1BL/1RS小麦变种SP4、莫迦小麦为供体材料,以杀雄剂途径培育的小麦强优势组合西杂1号和西杂5号杂交小麦新品种的母本西农Fp1和西农Fp2为受体材料,采用专一核置换回交转育方法,同时结合根尖细胞学镜检、复合引物PCR及SDS-PAGE和A-PAGE技术,进行不育基因rfv1定向转入的鉴定,旨在育成既携带rfv1不育基因,又具有西农Fp1和西农Fp2核遗传背景的黏类非1BL/1RS小麦雄性不育新保持系。结果如下:(1)根尖体细胞随体鉴定和复合引物PCR分析表明,该法不仅能准确鉴定出1BL/1RS纯合易位系,还可鉴定出1BL/1RS.1BL/1BSrfv1易位杂合体,两者结果一致。其中复合引物PCR更适合于回交后代中大量目标植株的筛选,为小麦雄性不育基因rfv1定向转移到1BL/1RS易位系提供了准确的鉴定方法与依据;(2)利用SDS-PAGE技术对供试材料进行高低分子量麦谷蛋白亚基的分析表明,在低分子量谷蛋白D亚基区存在莫迦小麦和SP4的特征带;而SP4的高分子量谷蛋白亚基区的6+8亚基,也可以作为示踪小麦雄性不育基因rfv1定向转移到非1BL/1RS易位系的特征亚基条带;(3)A-PAGE技术对醇溶蛋白的分析表明,在ω-醇溶蛋白区也发现莫迦小麦和SP4不同于西农Fp2的特异蛋白条带,也可作为示踪小麦雄性不育基因rfv1定向转移到非1BL/1RS易位系的特征蛋白条带。本研究成功地将小麦杂种优势利用中的杀雄剂法和三系法相结合,促进了小麦杂种优势利用新技术体系的建立。同时该方法亦可应用于黏类小麦雄性不育恢复基因Rfv1的定向转育,进而可实现小麦由生理型不育向遗传型不育的定向转化,以探索一套杂交小麦强优势组合多途径利用的新方法。
In order to utilize wheat heterosis in multiple pathways,a special backcross was performed to transform physiological male sterile-restoration system to genetic male sterile-restoration system directionally,and female parents ofhybrid wheat XZ1 and XZ5,which were bred by means of physiological male sterile-restoration system and had proved ofrobust heterosis,were used as recurrent parents crossing and backcrossing to Triticum spelta var.duhamelianum（for shortSP4）and T.macha var.subletschchumicum with male sterile gene rfv1 on short arm of 1B chromosome to breed the newnon-1BL/1RS maintainers of CMS.The results are as follows：（1）Both root-tip cytological identification and multiplexPCR makers exactly differentiated between homozygote and heterozygote of 1BL/1RS translocation,and multiplex PCRwas more suitable for the selection of large backcross progenies and provided an accurate method for tracing rfv1 gene indirectional transduction of non-1BL/1RS translocation;（2）The HMW（High Molecular Weight）and LMW（LowMolecular Weight）subunits analysis by SDS-PAGE revealed that the special subunit in D subunit area of SP4 and T.macha could trace rfv1gene in directional transduction.For SP4,6＋8 HMW subunit could be the tracer subunit;（3）Based on A-PAGE,the special gliadin band inω-gliadins area of SP4 and T.macha were found,which could be atracer gliadin band of rfv1gene in directional transduction.These results could be applied for directional transduction ofrestoring gene Rfv1 of wheat sterility lines with some Aegilops cytoplasms.
出处
《核农学报》
CAS
CSCD
北大核心
2010年第6期1124-1131,共8页
Journal of Nuclear Agricultural Sciences
基金
国家高技术研究发展计划(863计划)重大专项(2009AA101102)
高等学校博士学科点专项科研基金(20090204110024)
陕西省"13115"科技创新工程重大科技专项(2010ZDKG-68)
国家杨凌农业生物技术育种中心专项基金(99-1A)
西北农林科技大学拔尖人才支持计划项目
关键词
小麦
黏类雄性不育系
rfv1基因
定向转育
分子细胞学鉴定
wheat
wheat sterility lines with some Aegilops cytoplasms
rfv1 gene
directional transduction
molecular cytogenetical identification
