小麦-黑麦1BL/1RS易位系7-1抗纹枯病的分子细胞学鉴定  

Molecular Cytogenetic Identification of Wheat-rye(Triticum aestivumSecale cereale) 1BL/1R Translocation Line 7-1 with Sharp Eyespot Resistance

作　　者：梁邦平[1] 郝冬冬 刁慧珊 李家创 袁凤平[1] 李毛[1] 武军[1] 赵继新[1] 陈新宏[1] LIANG Bang-Ping HAO Dong-Dong DIAO Hui-Shan LI Jia-Chuang YUAN Feng-Ping LI Mao WU Jun ZHAO Ji-Xin CHEN Xin-Hong(College of Agronomy, Northwest A＆F University, Yangling 71210)

机构地区：[1]西北农林科技大学农学院,杨凌712100

出　　处：《农业生物技术学报》2018年第4期711-718,共8页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.31571650和No.31771785);国家重点研发计划(No.2017YFD0100701);陕西省重点实验室后补助项目(No.2016SZSj-20);西北农林科技大学唐仲英育种基金

摘　　要：黑麦(Secale cereale)是小麦(Triticum aestivum)的重要三级基因源,具有小麦改良所需要的多种优良性状。为丰富小麦杂交育种新的种质资源,利用墨西哥黑麦(2n=2x=14,RR)与普通优质小麦W770B(2n=6x=42,AABBDD)杂交,经过多年筛选,选出若干优良性状的衍生系,在小麦五叶期时用PDA培养基培养的带纹枯病菌的牙签,接菌到小麦芽鞘内,温室内培养,5周后鉴定纹枯病发病等级,计算病情指数,为了结果的准确性,经过多次牙签法鉴定,筛选出病情指数为PI=32.8%的抗病衍生系7-1。为明确衍生系7-1的遗传成分,本研究综合采用形态学、细胞遗传学、基因组原位杂交(genomic in situ hybridization,GISH)、特异序列扩增(sequence characterized amplified region,SCAR)分子标记、简单重复序列(simple sequence repeat,SSR)分子标记和醇溶蛋白分析。对7-1的根尖细胞和花粉母细胞的细胞遗传学观察表明7-1染色体结构和数目稳定2n=42=21Ⅸ;以黑麦DNA为探针的GISH分析观察表明7-1含有两个黑麦染色体臂;黑麦基因组SCAR标记分析表明,D15和P13LF/R能够在黑麦和7-1中扩增出黑麦特异条带,黑麦1RS SCAR标记分析表明,ω-sec-p1/ω-sec-p2、ω-sec-p3/ω-sec-p4和IB-267能够在黑麦和7-1中扩增出黑麦目的条带,说明7-1具有黑麦1RS染色体;筛选小麦每条染色体长短臂上的多对引物,在7-1中只有1BS上的引物Xgwm264和Xgwm11未能扩增出相应条带,其余染色体上的引物均扩增出了条带,说明7-1中缺失了小麦1BS染色体;醇溶蛋白分析表明7-1中扩增出了1RS黑麦碱条带,由此证实了小麦染色体1BS被黑麦染色体1RS所替换,该材料为小麦-黑麦1BL/1RS易位系。本研究中7-1为抗纹枯病的1BL/1RS易位系,为小麦纹枯病抗病育种提供了新的种质资源,拓宽了小麦育种材料。Rye（Secale cereale）, which contains excellent resistance and stress tolerance genes to against biotic and abiotic stress, is an important germplasm for wheat（Triticum aestivum） improvement. To enlarge the germlplasm resources for wheat breeding, the crosses between common wheat cv. W770 B（2 n=6 x=42,AABBDD） and rye（Secale cereale; 2 n=2 x=14, RR） via wide hybridization was conducted, and several derived lines with great characteristics were obtained after a long-time screening. Then, the toothpicks infected with Rhizoctonia cerealis which cultivated on freshly prepared potato-dextrose agar（PDA） were put in the leaf sheaths of these materials at the five-leaf stage of wheat plants. The plants after inoculation were cultivated in greenhouse for five weeks, and then identified the occurrence degrees and calculated the disease indexes.Derived line 7-1,with a sharp eyespot disease index of 32.8% was selected and identified by toothpick method through several years for improving the accuracy of the identification. In order to figure out the genetic constitution of 7-1, morphological method, cytogenetical method, genomic in situ hybridization（GISH）,sequence characterized amplified region（SCAR） markers, simple sequence repeat（SSR） markers and gliadin analysis were conducted in this study. The results of cytogenetics observation of root tip cells and pollen mother cells showed that the number and structure of chromosome of 7-1 were stable and the configuration of7-1 was 2 n=42=21 Ⅱ. GISH analyses using rye genomic DNA as probe indicated that 7-1 contained two chromosome arms of rye. The analysis of rye genome SCAR markers showed that D15 and P13 LF/R can amplify 900 bp around and 850 bp around rye specific bands both in rye and 7-1. And the analysis of 1RS SCAR markers of rye showed that ω-sec-p1/ω-sec-p2, ω-sec-p3/ω-sec-p4 and IB-267 amplified 1 100 bp around, 450 bp around and 300 bp around rye target bands both in rye and 7-1. Both analysis of rye genome SCAR markers and rye chromo

关 键 词：黑麦 基因组原位杂交(GISH) 特异序列扩增(SCAR) 简单重复序列(SSR) 1BL/1RS 

分 类 号：S512[农业科学—作物学]