落叶松-杨栅锈菌MlpMCM4蛋白和MlpHOG1蛋白互作关系初探  

A Preliminary Study on the Interaction Between MlpMCM4 and MlpHOG1 in Melampsora larici-populina

作　　者：杨冰 陈凯玥 李子晔 周显臻 于丹[1] YANG Bing;CHEN Kai-yue;LI Zi-ye;ZHOU Xian-zhen;YU Dan(College of Forestry,Northwest A&F University,Yangling 712100,Shaanxi,China)

机构地区：[1]西北农林科技大学林学院,陕西杨陵712100

出　　处：《西北林学院学报》2024年第1期100-107,共8页Journal of Northwest Forestry University

基　　金：国家自然科学基金(31870630)。

摘　　要：由落叶松-杨栅锈菌侵染引起的杨树叶锈病严重威胁杨树的健康生长。通过家族鉴定和系统发育分析确定落叶松-杨栅锈菌标准菌株ID 48743为MCM4直系同源基因,命名为MlpMCM4。以夏孢子cDNA为模板,运用RT-PCR技术,同源克隆获得中国菌株MlpMCM4基因CDS片段,称之为MlpMCM4(wh03),长度为2460 bp,编码819个氨基酸。结果表明,比对分析显示目的蛋白具有保守结构域,包括Walker A、Walker B和R-finger基序,以及MCM4类型锌指结构。亚细胞定位预测显示其定位在细胞核区域。利用基于分离泛素系统的酵母双杂交技术和萤火虫荧光素酶互补试验,没有检测到落叶松-杨栅锈菌中国菌株MlpMCM4蛋白和MlpHOG1蛋白能够相互作用,推断二者相互作用可能需要外源渗透压刺激。The poplar leaf rust caused by Melampsora larici-populina is a serious threat to the poplar growth.By family identification and the phylogenetic tree,it was found that ID 48743 from the Mlp referrence strain 98AG31 was the ortholog gene of MCM4 from Saccharomyces cerevisiae,so naming as MlpMCM4.The gene MlpMCM4(wh03)was cloned and sequenced from Mlp Chinese strain,using urediospore cDNA as template and the homology-based RT-PCR method.The coding sequence was 2460 bp,encoding 819 amino acids.The deduced MlpMCM4(wh03)protein contained the conserved domains,including Walker A,Walker B and R-finger motifs,and Zn finger domain based on alignment analysis.By means of the online bioinformatics tools,it was predicted that this protein was located at the nucleus region.No interaction between MlpMCM4(wh03)and MlpHOG1(wh03)was detected in both the split-ubiquitin yeast two hybrid system and the firefly luciferase complementary assay.It is inferred that the interaction may need the exogenous osmotic pressure stimulation.

关 键 词：落叶松-杨栅锈菌 MCM4基因 同源克隆 序列分析 蛋白互作 

分 类 号：S763.11[农业科学—森林保护学]