黑龙江省和海南省PWL基因家族在稻瘟病菌中的分布及变异  

Distribution and Variation of PWL Gene Family in Rice Magnaporthe oryzae from Heilongjiang Province and Hainan Province

作　　者：刘瑞[1] 赵羽涵 付忠举 顾欣怡 王艳霞 靳学慧[1] 杨莹 吴伟怀[2] 张亚玲[1] LIU RUI;ZHAO YuHan;FU ZhongJu;GU XinYi;WANG YanXia;JIN XueHui;YANG Ying;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2期264-274,共11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】了解不同稻瘟病菌(Magnaporthe oryzae)菌株群体中PWL基因家族的分布和变异特征,为研究不同稻瘟病菌群体的遗传多样性及特异性提供依据。【方法】通过参考NCBI中公布的无毒基因序列分别对PWL基因家族的启动子区和CDS区设计特异性引物共8对,对2020年采自黑龙江省和海南省不同地区的397个稻瘟病菌单孢分离菌株提取DNA,进行无毒基因的PCR扩增并通过琼脂糖凝胶电泳检测,从检测结果中分别选取囊括不同地区的代表菌株对扩增片段进行测序分析。测序结果与NCBI中相应无毒基因启动子区和CDS区的碱基与氨基酸序列进行比较分析。【结果】在PCR电泳检测结果中,PWL1在所有菌株中均未检测出;PWL2、PWL3和PWL4在黑龙江省和海南省中均扩增出特异性片段,说明这3种基因在两省中均有分布并分别存在不同的分布频率与变异类型。其中PWL2在黑龙江省和海南省中分布频率最高,分别为98.14%和100%;PWL3和PWL4在两省中的分布频率具有显著差异,这2种基因在黑龙江菌株中频率分别为89.30%和82.79%,而在海南菌株中均为5.49%。通过对无毒基因组合进行分析,结果显示,组合类型可分为6种,分别为PWL(f)、PWL2、PWL3、PWL2+PWL3、PWL2+PWL4、PWL2+PWL3+PWL4。其中黑龙江菌株含有所有组合类型,海南菌株仅含有2种,说明黑龙江菌株与海南菌株相比无毒基因型种类较为丰富。通过对PWL基因家族的PCR产物测序发现,PWL基因家族在启动子区和CDS区变异位点丰富,以点突变和缺失为主要变异类型划分为9种,且不同群体来源菌株的变异类型具有特异性和一致性。其中PWL2检测出5种变异类型PWL2-(1,2,3,4,5),碱基序列变化导致氨基酸序列发生错义突变;PWL3和PWL4分别检测出2种变异类型,分别为PWL3-(1,2)和PWL4-(1,2),均发生移码突变使后面氨基酸发生变化。【结论】不同群体来源的稻瘟病菌菌株中PWL基因家族的分布和【Objective】The objective of this study is to understand the distribution and variation characteristics of PWL gene family in different Magnaporthe oryzae strains, and to provide a basis for studying the genetic diversity and specificity of different M.oryzae strains.【Method】A total of 8 pairs of specific primers were designed for the promoter region and CDS region of the PWL gene family by referring to the avirulence gene sequence published in NCBI. DNA was extracted from 397 single spore isolates of M.oryzae collected from different regions of Heilongjiang Province and Hainan Province in 2020. PCR amplification of avirulence genes was conducted and the genes were detected by agarose gel electrophoresis. Representative strains from different regions were selected from the test results to sequence and analyze the amplified fragments. The sequencing results were compared with the base and amino acid sequences of the corresponding avirulence gene promoter region and CDS region in NCBI.【Result】In the results of PCR electrophoresis, PWL1 was not detected in all strains. The specific fragments of PWL2, PWL3 and PWL4 were amplified in Heilongjiang Province and Hainan Province, indicating that these three genes were distributed in both provinces and had different distribution frequencies and variation types. Among them, PWL2 had the highest distribution frequency in Heilongjiang Province and Hainan Province, which were 98.14% and 100%, respectively. The distribution frequencies of PWL3 and PWL4 in the two provinces were significantly different. The frequencies of these two genes in Heilongjiang strains were 89.30% and 82.79%, respectively, while in Hainan strains were 5.49%. Through the analysis of avirulence gene combinations, the results showed that the combination types could be divided into six types, namely PWL(f), PWL2, PWL3, PWL2+PWL3, PWL2+PWL4, PWL2+PWL3+PWL4. Among them,Heilongjiang strains contained all combination types, and Hainan strains contained only two, indicating that Heilongjiang strains w

关 键 词：稻瘟病菌 PWL基因家族 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]