菲律宾稻瘟病菌生理小种中AVR-Pita及其同源基因的序列与功能分析

doi:10.16409/j.cnki.2095-039x.2018.03.006

摘要/Abstract

摘要： 根据已克隆的AVR-Pita及其同源基因序列设计4对引物，对来自菲律宾的74个稻瘟病单孢菌株DNA进行PCR扩增和序列分析，研究了与水稻抗病基因Pi-ta互作的稻瘟病菌无毒基因AVR-Pita及其同源基因序列多态性与进化关系。结果显示，在42个菌株中扩增出AVR-Pita1基因目的条带，31个菌株中扩增出AVR-Pita3基因目的条带，55个菌株中扩增出AVR-Pita4基因目的条带，但所有供试菌株均未扩增出AVR-Pita2基因目的条带。对PCR产物进行测序分析，发现AVR-Pita1基因在菲律宾的生理小种中存在序列多态性，可划分为5个单倍型，共有10个多态性位点；AVR-Pita3和AVR-Pita4基因序列较保守，序列比对后没有发现多态性。对不同单基因系接种鉴定，发现除第3种AVR-Pita1单倍型外，含有AVR-Pita1单倍型1、2、4及5的生理小种均不能与Pi-ta基因互作，从而使Pi-ta单基因系IRBLta-CT2在接种后表现为感病。以上试验结果说明AVR-Pita1基因变异性较强，且序列改变后会导致基因功能的变化。

关键词: 稻瘟病菌, AVR-Pita及其同源基因, 序列分析, 功能分析

Abstract: For controlling rice blast and providing theory for breeding and selecting resistant rice variety, we studied the sequence polymorphism and function of avirulence gene AVR-Pita homologs. According to the cloned AVR-Pita homologs, we designed 4 primer pairs, selected 74 isolates from Philippine and distracted their DNA as template for PCR. The results showed that 42 isolates had the band of AVR-Pita1 gene, 31 isolates had the band of AVR-Pita3 gene, and 55 isolates had the band of AVR-Pita4 gene. However, no isolates showed the band of AVR-Pita2 gene. Sequences analysis of PCR products showed there were 5 haplotypes among AVR-Pita1 gene with variation in 10 base sites, while there was no polymorphic base site in AVR-Pita3 and AVR-Pita4 genes, indicating both genes were conservative. Inoculation tests demonstrated that IRBLta-CT2 was susceptible after inoculated with the haplotype 1, 2, 4 and 5, except haplotype 3. In conclusion, AVR-Pita1 gene is of strong variation, and its function will change after gene sequence has changed.

Key words: rice blast fungus, AVR-Pita gene family, sequence analysis, function analysis

中图分类号:

S435.11

甘玉姿, 肖贵, 邓启云, 吴俊, 柏斌, 卢向阳, 周波. 菲律宾稻瘟病菌生理小种中AVR-Pita及其同源基因的序列与功能分析[J]. 中国生物防治学报, 2018, 34(3): 488-498.

GAN Yuzi, XIAO Gui, DENG Qiyun, WU Jun, BAI Bin, LU Xiangyang, ZHOU Bo. The Sequence and Function Analysis of AVR-Pita Gene Family of Rice Blast Fungus, Magnaporthe oryzae in Philippines[J]. journal1, 2018, 34(3): 488-498.

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