源于广8 A杂交稻组合的稻瘟病菌无毒基因型分析  

Analysis of Magnaporthe oryzae Avirulent Genes in the Infected Hybrid Rice Combinations Derived from a Sterile Line of Guang 8 A

作　　者：汪文娟[1] 苏菁[1] 杨健源[1] 韦小燕[1] 陈凯玲[1] 陈珍[1] 陈深[1] 朱小源[1] WANG Wen Juan;SU Jing;YANG JianYuan;WEI XiaoYan;CHEN KaiLing;CHEN Zhen;CHEN Shen;ZHU XiaoYuan(Plant Protection Research Institute,Guangdong Academy of Agricultural Sciences/Guangdong Provincial Key Laboratory of High Teehnology for Plant Protection,Guangzhou 510640)

机构地区：[1]广东省农业科学院植物保护研究所/广东省植物保护新技术重点实验室,广州510640

出　　处：《中国农业科学》2018年第24期4633-4646,共14页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2016YFD0300707);广东省现代农业产业技术体系(2017LM1075);广东省科技计划项目(2015B020231003;2016B090918116;2016A020210026);国家现代农业产业技术体系建设专项(CARS-01-32)

摘　　要：【目的】分析源于广8 A杂交稻组合的稻瘟病菌(Magnaporthe oryzae)无毒基因型,为华南不同主栽抗病水稻品种的合理布局提供参考依据。【方法】利用抗稻瘟病单基因系,对稻瘟病菌单孢分离菌株进行致病型测定;并根据已克隆的且与稻瘟病菌致病性相关的8个无毒基因功能性分子标记进行分离菌株的无毒基因型分析。选取分离自广8 A杂交稻组合的27个稻瘟病菌单孢菌株,提取各菌株的菌丝体DNA作为模板,进行PCR扩增及琼脂糖凝胶电泳检测。并对其中的7个稻瘟病菌菌株进行相应无毒基因全长或CDS区域的PCR产物测序,将测序序列与相应无毒菌株的无毒基因序列进行比较分析。【结果】所测定的菌株对其中的5个单基因系IRBLkh-K3(Pikh)、IRBLb-B(Pib)、IRBLz5-CA(Pi2)、NIL-e1(Pi50)和IRBL9-W(Pi9)表现高的无毒性频率(>85%),所有菌株对单基因系IRBL9-W(Pi9)和NIL-e1(Pi50)都表现无毒;这些菌株对其中的7个单基因系表现较低的无毒性频率(<20%),包括IRBLks-F5(Piks)、IRBLa-A(Pia)和IRBL19-A(Pi19)等,表明分离自广8 A杂交稻组合的稻瘟病菌对不同的单基因系表现出不同的致病型。所有菌株都被检测出含无毒基因AvrPi9和AvrPik,并可检测到无毒基因AvrPik的2种类型:AvrPik-D型和AvrPik-E型,但未检测出无毒基因Avr1-CO39、AvrPii和AvrPia。在部分菌株中能检测到无毒基因AvrPiz-t、AvrPib和AvrPita预期大小的产物,但在所检测的菌株中均以较低的频率与不同的突变类型出现。对GD13-621、GD14-349与GD15-291等7个菌株分别进行相应无毒基因的PCR产物测序,AvrPi9、AvrPita及AvrPiz-t的基因序列与其各自无毒菌株的无毒基因序列完全一致,表明分离菌株中这3个无毒基因的基因结构相对稳定。测序菌株的AvrPib基因序列与已报道的AvrPib无毒菌株的序列在基因的上游区域存在2个SNP的差异及3个连续碱基的插入,具有较丰富的多样性;6个测序菌株的AvrP【Objective】 The objective of this study is to analyze the avirulent genotypes of Magnaporthe oryzae derived from the hybrid rice combinations of Guang 8 A, and to provide a reference for rational distribution of cultivars with different blast resistant genes in South China. 【Method】 A set of single-spore strain was subjected to pathotype analysis using blast monogenic differential lines. According to the functional markers of 8 Avr genes, which had been cloned and were associated with the rice blast pathogenicity, the haplotypes of Avr genes were analyzed. The DNA, which extracted from the 27 strains of rice blast fungi, was used as a template for PCR amplification. The PCR products of avirulent full gene or CDS region were analyzed by agarose gel electrophoresis and sequencing, and their sequences were compared to corresponding avirulent genes【.Result】The tested strains showed high frequency(>85%) of avirulence to the 5 monogenic differential lines, such as IRBLkh-K3(Pikh), IRBLb-B(Pib), IRBLz5-CA(Pi2), NIL-e1(Pi50) and IRBL9-W(Pi9), all strains were avirulent to IRBL9-W(Pi9) and NIL-e1(Pi50). These strains showed relatively low frequency of avirulence(<20%) to the 7 monogenic differential lines, such as IRBLks-F5(Piks), IRBLa-A(Pia), IRBL19-A(Pi19) and so on, indicating that the strains from the combination of Guang 8 A showed different pathotypes to different rice blast-resistant lines. AvrPi9 and AvrPik fragments were almost present in all strains, and two haplotypes(AvrPik-D and AvrPik-E) of AvrPik were identified. But none of Avr1-CO39, AvrPii or AvrPia was amplified in any strain. The expected size products of AvrPiz-t, AvrPib and AvrPita could be detected in some strains, but they all appeared in lower frequency and different mutation types in the tested strain. Amplicon sequencing of 7 strains(GD13-621, GD14-349, GD15-291, et al.) revealed that the sequences of AvrPi9, AvrPita and AvrPiz-t were identical to those of the respective avirulent strains. This result indicated that these 3 Avr gene

关 键 词：杂交稻组合 广8 A 稻瘟病菌 无毒基因 抗稻瘟病单基因系 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]