黑龙江省和海南省稻瘟病菌中AVR-Pita家族的分布及变异分析  

Distribution and Variation Analysis of AVR-Pita Family in Magnaporthe oryzae from Heilongjiang Province and Hainan Province

作　　者：刘瑞[1] 赵羽涵 顾欣怡 王艳霞 靳学慧[1] 吴伟怀[2] 张亚玲[1] LIU RUI;ZHAO YuHan;GU XinYi;WANG YanXia;JIN XueHui;WU WeiHuai;ZHANG YaLing(College of Agronomy,Heilongjiang Bayi Agricultural University/Heilongjiang Plant Resistance Research Center,Daqing 163319,Heilongjiang;Environment and Plant Protection Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101)

机构地区：[1]黑龙江八一农垦大学农学院/黑龙江省植物抗性研究中心,黑龙江大庆163319 [2]中国热带农业科学院环境与植物保护研究所,海口571101

出　　处：《中国农业科学》2023年第3期466-480,共15页Scientia Agricultura Sinica

基　　金：国家自然科学基金(U20A2025);黑龙江省农垦总局科技攻关项目(HKKYZD190205);黑龙江八一农垦大学科研启动项目(XDB201802,XDB201605)。

摘　　要：【目的】通过检测黑龙江省和海南省不同稻瘟病菌(Magnaporthe oryzae)菌株群体中AVR-Pita家族的分布情况和变异特征,了解其变异类型的致病表型,为区域内抗病种质的筛选与选育提供参考。【方法】通过参考NCBI中公布的AVR-Pita家族序列分别对启动子区和CDS区设计特异性引物共6对,对2020年采自黑龙江省和海南省不同地区的397个稻瘟病菌单孢菌株提取DNA,进行PCR扩增。通过电泳检测结果,分别选取囊括不同地区的代表菌株对扩增片段进行测序。测序结果与NCBI中相应的碱基与氨基酸序列进行比较分析,并利用水稻抗性单基因系,对不同变异类型的稻瘟病菌菌株进行无毒功能验证。【结果】在PCR电泳检测结果中,黑龙江省稻瘟病菌菌株携带所有待测基因,分布广泛且检出频率较高;而海南省稻瘟病菌菌株中仅携带AVR-Pita1和AVR-Pita2,并以低频率集中存在。无毒基因组成分析结果显示,黑龙江稻区的稻瘟病菌菌株复杂多样,携带的基因型种类较海南菌株丰富。PCR产物测序检测结果显示,AVR-Pita家族以点突变、插入和缺失为主要变异类型划分为19种,且不同稻瘟病菌群体来源的菌株,其变异类型具有特异性。AVR-Pita1检测出10种变异类型,其中AVR-Pita1-(1-5)为黑龙江省稻瘟病菌菌株独有的变异类型,AVR-Pita1-(6-10)为海南省稻瘟病菌菌株独有。对这10种变异类型经功能验证后发现,无毒功能均已丧失。AVR-Pita2检测出8种变异类型,其中AVR-Pita2-(1-4)为黑龙江省稻瘟病菌菌株独有,AVR-Pita2-(5-8)为海南省稻瘟病菌菌株独有。经功能验证后发现,无毒功能均已丧失。AVR-Pita3在黑龙江省仅检测出1种变异类型AVR-Pita3-1。【结论】黑龙江和海南稻瘟病菌群体中AVR-Pita家族均为突变后的等位基因型存在,经致病性鉴定后,检测出的所有突变类型均不能被相对应抗性基因Pi-ta和Pi-ta2所识别,表现为感病。因此,抗【Objective】By detecting the distribution and variation characteristics of the AVR-Pita family in different Magnaporthe oryzae strains from Heilongjiang Province and Hainan Province,the pathogenic phenotypes of the variation types were understood,which provided a reference for the screening and breeding of disease-resistant germplasm in the region.【Method】Six pairs of specific primers were designed for the promoter region and CDS region by referring to the AVR-Pita family sequence published in NCBI.DNA was extracted from 397 single spore strains of M.oryzae collected from different regions of Heilongjiang Province and Hainan Province in 2020,and PCR amplification was performed.By electrophoresis test results,representative strains from different regions were selected to sequence the amplified fragments.The sequencing results were compared with the corresponding base and amino acid sequences in NCBI,and the avirulence function of different variation types of M.oryzae strains was verified by using rice resistant single gene lines.【Result】In the PCR electrophoresis results,the M.oryzae strains in Heilongjiang Province carried all the genes to be tested,which were widely distributed and had a high detection frequency.However,only AVR-Pita1 and AVR-Pita2 were carried by M.oryzae strains in Hainan Province,and concentrated in low frequency.The results of avirulence gene composition analysis showed that the strains of M.oryzae in Heilongjiang were complex and diverse,and the genotypes were more abundant than those in Hainan.The sequencing results of PCR products showed that the AVR-Pita family was divided into 19 types with point mutation,insertion and deletion as the main variation types,and the variation types of strains from different M.oryzae populations were specific.Ten variation types were detected in AVR-Pita1,among which AVR-Pita1-(1-5)was a unique variation type of M.oryzae strains in Heilongjiang Province,and AVR-Pita1-(6-10)was a unique variation type of M.oryzae strains in Hainan Province.After f

关 键 词：稻瘟病菌 AVR-Pita家族 变异 无毒基因 黑龙江省 海南省 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]