小麦-黑麦1RS-1BS/1BL小片段易位系抗赤霉病鉴定及分子细胞遗传学分析  

Identification and Molecular Cytogenetic Analysis of Wheat-rye(Triticum aestivum-Secale cereale)1RS-1BS/1BL Small Fragment Translocation Lines Against Fusarium Head Blight

作　　者：吕婷婷 张珍悦 姚晓妮 张寒冰 李家创 刘淑会[1] 杨群慧[1] 武军[1] 陈新宏[1] LV Ting-Ting;ZHANG Zhen-Yue;YAO Xiao-Ni;ZHANG Han-Bing;LI Jia-Chuang;LIU Shu-Hui;YANG Qun-Hui;WU Jun;CHEN Xin-Hong(Shaanxi Key Laboratory of Plant Genetic Engineering Breeding,College of Agronomy,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学农学院/陕西省植物遗传工程育种重点实验室,杨凌712100

出　　处：《农业生物技术学报》2020年第7期1193-1202,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金项目(31571650;31771785);国家重点研发计划项目(2017YFD0100701);陕西省农业领域重点项目(2018ZDXM-NY-006)。

摘　　要：赤霉病(Fusarium head blight,FHB)是小麦的主要病害之一,对小麦(Triticum aestivum;2n=6x=42,AABBDD)正常生长和产量造成了严重的威胁。黑麦(Secale cereal;2n=2x=14,RR)具有发达的根系以及较强的抗逆性与抗病性,是改良小麦性状的重要遗传资源。为了提高小麦抗赤霉病的能力,本实验通过墨西哥黑麦与普通小麦W770B杂交,经多代选育获得了一批能够稳定遗传的衍生后代。在小麦扬花期,采用单花滴注法(single flower inoculation,SFI)对亲本及其衍生后代进行赤霉病抗性鉴定,结果显示亲本黑麦对赤霉病表现中抗,W770B表现高感,衍生后代12-5-1表现中抗。进一步通过基因组原位杂交(genome in situ hybridization,GISH)、荧光原位杂交(fluorescence in situ hybridization,FISH)、特异序列扩增分子标记(sequence characterized amplified regions,SCAR)、简单重复序列分子标记(simple sequence report,SSR)对中抗赤霉病的衍生后代12-5-1进行形态学及细胞遗传学的鉴定与分析。结果表明:12-5-1根尖细胞中含有42条染色体2n=42;花粉母细胞减数第一次分裂中期,染色体能正常配对形成21个二价体2n=42=21Ⅱ,并且能正常分离。以黑麦基因组为探针对12-5-1进行GISH鉴定,观察到小麦的2条染色体端部有黄绿色信号,说明小麦染色体与黑麦染色体发生了易位;FISH鉴定表明12-5-1中易位染色体为小麦的1BS与黑麦的1RS发生了易位;因此推断小麦与黑麦杂交发生了染色体小片段易位。利用黑麦特异性标记AF1/AF4与1RS的SCAR标记进行鉴定,结果只在黑麦和衍生后代12-5-1中扩增出相应的条带;SSR分析结果显示小麦1BS染色体上的3对引物(Xgwm11、Xgwm31、Xgwm550)不能在12-5-1中扩增出目标条带,其余引物均能扩增出相应的目标条带,表明12-5-1是一个1RS-1BS/1BL的小麦-黑麦小片段易位系。该抗病衍生后代的创制为小麦抗赤霉病育种提供了新的种质资源。Fusarium head blight(FHB)is one of the main diseases which caused a serious threat to the normal growth and yield formation of wheat(Triticum aestivum;2n=6x=42,AABBDD).Rye(Secale cereal;2n=2x=14,RR)with developed root system,strong stress resistance and disease resistance,is an important genetic resource for improving wheat traits.For the purpose of improving the FHB resistance in wheat,Mexican rye and common wheat W770B were hybridized to obtain several derived lines with genetic stability through multi-generation breeding.At the blooming stage of wheat,single flower inoculation(SFI)was used to identify the parents and progenies resistance to FHB.The results showed that the parents rye had moderate resistance to FHB whereas W770B showed high sensitivity,and the 12-5-1 derived line showed moderate resistance.Morphological and cytogenetic identification and analysis were conducted in 12-5-1 derived line by genomic in situ hybridization(GISH),fluorescence in situ hybridization(FISH)sequence amplification,sequence characterized amplified regions(SCAR),simple sequence repeats(SSR)molecular markers.The results showed that the root tip cell of 12-5-1 contained 42 chromosomes(2n=42).In the first meiotic metaphase of pollen mother cell,there is a proper chromosome pairing forming 21 bivalents(2n=42=21Ⅱ),and normal separation.GISH identification was performed on 12-5-1 using rye genome as a probe.Yellow and green signals were observed at the ends of two chromosomes of wheat,indicating that there is a translocation between wheat chromosome and rye chromosome.FISH results showed that the translocation of 1BS of wheat and 1RS of rye occurred in 12-5-1.Therefore,it is inferred that the hybridization of wheat and rye resulted in translocation of small chromosome segments.Moreover,Rye specific marker AF1/AF4 and SCAR marker of 1RS were used for identification and the corresponding bands were amplified in rye and 12-5-1 derived line.SSR analysis showed that three pairs of primers(Xgwm11、Xgwm31 and Xgwm550)on the 1BS chromoso

关 键 词：小麦 黑麦 赤霉病 易位系 基因组原位杂交(GISH) 荧光原位杂交(FISH) 

分 类 号：S512[农业科学—作物学]