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马铃薯晚疫病菌黑龙江菌株中12个RXLR类分泌蛋白基因的克隆与初步功能分析

发表时间:2014-10-15  浏览量:1655  下载量:577
全部作者: 王娇,董冉,吴思思,丁新华,储昭辉
作者单位: 山东农业大学植物保护学院,山东省农业微生物重点实验室
摘 要: 以我国黑龙江地区分离菌株为研究对象,参考基因组测试菌株T30-4的序列信息,随机选取12个RXLR基因设计引物克隆基因。12个候选基因在黑龙江菌株中均成功克隆到DNA片段,与参考菌株T30-4的序列相比,其中5个基因DNA序列完全相同,另外7个同源基因的序列存在DNA变异,基因突变频率为58.33%. 其中,6个同源基因的序列变异导致推导氨基酸序列产生1~5个位点的突变。利用本氏烟(Nicotiana benthamiana)瞬时表达系统对12个基因进行初步功能分析,全部基因均未产生自激活反应,排除激发子功能;其中,PITG-21645.2的基因与INF1在本氏烟上共表达能抑制INF1引起过敏性坏死反应(hypersensitive response, HR)的产生。PITG-21645.2与T30-4菌株中等位基因相比只有1个氨基酸的差别,点突变分析发现该氨基酸的变化并不影响抑制功能。这为探究我国菌株RXLR分泌蛋白参与致病机理的功能研究提供了初步实验依据。
关 键 词: 植物病理学;马铃薯;致病疫霉菌;分泌蛋白;细胞程序性死亡;抑制子
Title: Cloning and preliminary functional analysis of 12 genes encoding RXLR secreted proteins from Phytophthora infestans Heilongjiang isolates
Author: WANG Jiao, DONG Ran, WU Sisi, DING Xinhua, CHU Zhaohui
Organization: Shandong Provincial Key Laboratory of Agricultural Microbiology, Plant Protection College, Shandong Agricultural University
Abstract: In this study, 12 RXLR genes were randomly selected as candidates to clone their homologues from Heilongjiang isolates (HLJ isolates) according to sequenced genome of T30-4 strain. Total 12 candidates were successfully amplified from HLJ isolates. Five out of 12 candidates shared the same sequence with their homologues from T30-4 strain. Seven candidates were identified to harbor nucleotide changes which represented the gene mutation ratio as 58.33%. Six out seven mutated genes were found to result in one to five amino acids substitution. Based on transient expression system in Nicotiana benthamiana, the function of total 12 RXLR effectors were also have been preliminary analysed. None of them could be activated hypersensitive response (HR) when they were expressed in N.benthamiana. However, when they were co-expressed with INF1 in N.benthamiana, only PITG-21645.2 was identified to suppress the HR triggered by INF1. There has one amino acid substitution between HLJ and T30-4 isolates in PITG-21645.2. And the results indicated that this point mutation will not influence the suppressor ability. These results have provided the initial experimental basis for exploring the pathogenicity of RXLR effectors with our national isolates of Phytophthora infestans.
Key words: plant pathology; potato; Phytophthora infestans; secretion protein; programmed cell death; suppressor
发表期数: 2014年10月第19期
引用格式: 王娇,董冉,吴思思,等. 马铃薯晚疫病菌黑龙江菌株中12个RXLR类分泌蛋白基因的克隆与初步功能分析[J]. 中国科技论文在线精品论文,2014,7(19):1930-1936.
 
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