Pi9基因分子标记辅助选择改良水稻不育系丰源A的稻瘟病抗性  

Improving Blast Resistance of CMS Line Fengyuan A by Pi9-based Marker-assisted Selection Strategy

作　　者：吴婷婷 陈海龙[1] 黄俊[1] 梁毅[1] 张婷 李博文 赖怡帆 邹玉莹 车凡昊 吴俊[4] 刘金灵[1,2,3] 刘雄伦 WU Tingting;CHEN Hailong;HUANG Jun;LIANG Yi;ZHANG Ting;LI Bowen;LAI Yifan;ZOU Yuying;CHE Fanhao;WU Jun;LIU Jinling;LIU Xionglun(College of Agronomy,Hunan Agricultural University,Changsha 410128,China;Crop Gene Engineering Key Laboratory of Hunan Province,Changsha 410128,China;Hunan Provincial Key Laboratory of Rice and Rapeseed Breeding for Disease Resistance,Changsha 410128,China;Hunan Hybrid Rice Research Center,Changsha 410125,China)

机构地区：[1]湖南农业大学农学院,湖南长沙410128 [2]作物基因工程湖南省重点实验室,湖南长沙410128 [3]水稻油菜抗病育种湖南省重点实验室,湖南长沙410128 [4]湖南杂交水稻研究中心,湖南长沙410125

出　　处：《华北农学报》2021年第5期191-197,共7页Acta Agriculturae Boreali-Sinica

基　　金：国家转基因重大专项(2016ZX08001-002)。

摘　　要：为改良水稻不育系丰源A的稻瘟病抗性,以籼稻品系75-1-127作为广谱持久抗稻瘟病基因Pi9的供体亲本,先后以三系保持系丰源B和不育系丰源A为受体亲本,利用Pi9基因内共显性标记ClonDF_(1) R_(1)开展MAS连续回交育种。利用来自国内外不同稻区的33份稻瘟菌菌株进行温室内人工接种,分析丰源A、丰源B、75-1-127的抗菌谱,75-1-127的抗性频率为87.9%;而丰源A和丰源B的抗性频率均仅为33.3%。田间病圃抗性鉴定表明,75-1-127高抗苗瘟,而丰源B和丰源A高感苗瘟。结果表明,含有Pi9基因的75-1-127抗谱广、抗性水平高,在稻瘟病抗性育种中具有很大的应用前景;而丰源A和丰源B抗谱窄、抗性水平低,需要改良。根据Pi9基因序列信息开发了一个共显性分子标记ClonDF_(1) R_(1),它从75-1-127基因组中扩增出一条320 bp的条带,而丰源B和丰源A基因组的扩增产物约450 bp。利用ClonDF_(1) R_(1)先后开展丰源B、丰源A稻瘟病抗性改良的MAS育种实践,获得了13个含Pi9纯合等位基因的改良抗病保持系丰源B-Pi9,以及11个含Pi9纯合等位基因的改良抗病不育系丰源A-Pi9。经过连续11 a的MAS回交育种实践,育成1个高抗稻瘟病、不育性稳定、柱头外露率高、农艺性状和产量性状优良的新三系不育系丰源A-Pi9-5,实现了丰源A和丰源B稻瘟病抗性的定向改良,为三系杂交水稻育种应用创制了新的亲本材料。To improve blast resistance of the CMS line Fengyuan A,a PCR-based co-dominant marker,ClonDF_(1) R_(1),was developed according to sequence information of the broad-spectrum and durable blast resistance gene Pi9,and was employed in marker-assisted selection breeding practice.Resistance responses to 33 tested M.oryzae isolates showed difference between the Pi9 donor 75-1-127 and the receptors Fengyuan A and its maintainer Fengyuan B,presenting 87.9%and 33.3%resistance frequency respectively in greenhouse inoculation assay.75-1-127 showed high-level seedling blast resistance but highly susceptible for two receptors in field nursery.These results showed that the Pi9 gene was promising in rice blast resistance breeding,and blast resistance of two receptors needs to be improved urgently.The ClonDF_(1) R_(1) was developed based on DNA sequence information of the Pi9 gene,this codominant marker primers amplified a 320 bp band from the donor genome while PCR product from two receptors was nearly 450 bp.The codominant molecular marker ClonDF_(1) R_(1) was used in MAS backcross breeding practice for improving blast resistance of Fengyuan B and Fengyuan A respectively.Thirteen maintainer lines Fengyuan B-Pi9,and eleven CMS lines Fengyuan A-Pi9,carrying the Pi9 homozygous alleles and showing high-level seedling blast resistance,were developed.One improved CMS line,Fengyuan A-Pi9-5,was bred through eleven-year consecutive MAS practice.Fengyuan A-Pi9-5 showed high-level seedling blast resistance,stable male sterility performance,high exserted stigma percentage,excellent agronomic and yield traits.Thus,we achieved the goal to directionally improve blast resistance of rice CMS line Fengyuan A and its maintainer Fengyuan B,and created a set of new parents for three-line method hybrid rice breeding application.

关 键 词：水稻 稻瘟病抗性改良 分子标记辅助选择 丰源A Pi9 

分 类 号：S435.11[农业科学—农业昆虫与害虫防治] S334.2[农业科学—植物保护]