中文摘要：

【目的】Pbs2是MAPK信号途径HOG-MAPK通路的重要成员之一,在植物病原菌渗透压调节方面发挥着重要作用。橡胶树白粉病菌（Oidium heveae）是专性寄生菌,论文借助橡胶树炭疽病菌（Colletotrichum gloeosporioides）,研究橡胶树白粉病菌OhPbs2的功能。【方法】采用同源克隆方法,分别以橡胶树白粉病菌基因组DNA和c DNA为模板,PCR扩增OhPbs2;利用生物信息学分析该基因的结构域,对该基因和其他真菌的7个同源蛋白序列进行系统进化分析,并利用MEGA6中最大简约法构建系统进化树来进一步分析鉴定该基因;利用同源重组和原生质体转化技术,将OhPbs2转化到缺失Pbs2的橡胶树炭疽菌突变体ΔCgPbs2中;在PDA＋1.5 mol·L-1山梨醇的平板上筛选转化子,同时提取转化子的基因组作为模板,用橡胶树白粉病菌OhPbs2的引物对进行PCR鉴定,选择正确的转化子ΔCgPbs2＋OhPbs2进行后续表型测定;在不同培养条件下,比较ΔCgPbs2、ΔCgPbs2＋OhPbs2和野生型菌株生长状态,并在橡胶树叶片接菌检测3个菌株致病力。【结果】OhPbs2基因序列全长1 927 bp,c DNA序列全长1 860 bp,含有一个内含子,编码一个619个氨基酸的蛋白;生物信息学分析表明该蛋白具有与Cg Pbs2相同的S_TKc功能域,构建系统进化树发现橡胶树白粉病菌Pbs2蛋白与烟曲霉（Aspergillus fumigatus）Pbs2蛋白亲缘关系最近,相似度为55%,与橡胶树炭疽病菌的相似度为49%,与粗糙脉孢菌（Neurospora crassa）、稻瘟菌（Magnaporthe oryzae）、酿酒酵母（Saccharomyces cerevisiae）、禾谷镰孢（Fusarium graminearum）的Pbs2蛋白具有较近的亲缘关系,相似度分别为54%、53%、53%、50%;ΔCgPbs2＋OhPbs2菌株能在PDA＋1.5 mol·L-1山梨醇培养基上长出白色菌落,ΔCgPbs2菌株不能生长,并且测序结果显示OhPbs2已成功转入ΔCgPbs2;ΔCgPbs2＋OhPbs2在MM培养基上菌落呈白色,气生菌丝较短,不同于wild type菌株。在含有不同浓度Na Cl

英文摘要：

[ Objective] Pbs2 is one of the important members of HOG-MAPK pathway in MAPK signaling pathway, and plays an important role in osmotic regulation of plant pathogens. Oidium heveae is an obligate parasite. In this paper, the function and effect of Pbs2 of O. heveae were studied by using Colletotrichum gloeosporioides. [Method] The homologous cloning method was used to amplify the OhPbs2 by using the genomic DNA and cDNA as template. The domain of this gene was analyzed by bioinformatics. Phylogenetic analysis of seven homologous protein sequences of other fungi and OhPbs2 was conducted, and the phylogenetic tree was constructed by the maximum parsimony method in MEGA6 to further analyze and identify this gene. Using homologous recombination and protoplast transformation, OhPbs2 was transformed into the ACgPbs2 of C. gloeosporioides. The transformant was screened on PDA＋1.5 mol.L-1 sorbitol. At the same time, the genome of the transformant was extracted as a template and identified with the primer pairs of OhPbs2, and the correct transformant △CgPbs2＋OhPbs2 was selected for subsequent phenotypic determination. The growth state of ACgPbs2, ACgPbs2＋OhPbs2 and wild type strains was compared under different culture conditions. And the pathogenicity of the three strains was detected by inoculating the leaves of the rubber tree. [ Result ] The full-length of the OhPbs2 is 1 927 bp, cDNA of the OhPbs2 is 1 860 bp, and contains an intron that encodes a 619 amino acids protein. Bioinformatics analysis showed that the protein had the same S_TKc domain as CgPbs2. The phylogenetic tree showed that the Pbs2 protein was closely related to Aspergillus fumigatus Pbs2 protein, and the similarity was 55%. And the similarity to Colletotrichum gloeosporioides was 49%, also it was close to Pbs2 protein of Neurospora crassa, Magnaporthe oryzae, Saccharomyces cerevisiae and Fusarium graminearum, the similarities were 54%, 53%, 53% and 50%, respectively. ACgPbs2＋OhPbs2 strain could grow white colonies on PDA＋1.5 mol.L-1 sorbi