Pseudomonas protegens H78中抗真菌剂藤黄绿菌素的合成及其基因簇分析  

The Pyoluteorin (Plt) Biosynthesis and Gene Cluster Analysis in Pseudomonas protegens H78

作　　者：杨国环 王正[1] 吴玲玉[1] 张雪洪[1] 黄显清[1] 

机构地区：[1]上海交通大学生命科学技术学院微生物代谢国家重点实验室,上海200240

出　　处：《基因组学与应用生物学》2015年第4期680-684,共5页Genomics and Applied Biology

基　　金：国家自然科学基金(No.31270083;31470196)资助

摘　　要：假单胞菌株H78是本实验室从油菜根际分离得到的一株生防细菌。通过16S r RNA及藤黄绿菌素(pyoluteorin,Plt)合成基因plt C基因(编码聚酮合成酶)保守区段进行PCR测序,初步确定H78菌株可能属于P.protegens。本研究旨在进一步克隆H78菌株的Plt生物合成基因簇,分析其Plt合成特性以及H78菌株的分类地位。对假单胞菌株H78进行全基因组de novo测序;使用blast工具,分析假单胞菌株H78与P.protegens Pf-5、CHA0、P.aeruginosa M18、LESB58中Plt合成基因簇的同源性;使用KMB培养基对菌体进行发酵,测定菌体生长,对Plt合成进行HPLC测定。假单胞菌H78的Plt合成基因簇与其它4个菌株都存在保守的基因组成与布局,在Plt合成基因编码氨基酸序列上,H78与CHA0、Pf-5菌株的一致性高达99%~100%,其中与CHA0同源性更高;而与铜绿假单胞菌M18及LESB58菌株的一致性相对较低,为52%~88%。在KMB培养基中,假单胞菌株H78中累积合成Plt超过30滋g/m L。H78菌株属于P.proteg...The biocontrol Pseudomonas strain H78, which was isolated from rape rhizosphere has tentatively been identified as P. protegens by PCR amplification and sequencing of the 16 S r RNA gene and the pyoluteorin(Plt)biosynthetic gene plt C. This study aims to further characterize the Plt biosynthetic gene cluster and investigate the characteristics of Plt production and the taxonomic position of H78 strain. First, the whole genome de novo sequencing of H78 was conducted. Then BLAST was used to assess the homology level of the Plt gene cluster between H78 and other four strains including P. protegens CHA0, Pf-5, P. aeruginosa M18 and LESB58. At last,we carried out the dynamics analysis of cell growth and Plt production of H78 in the KMB media. There is conservative gene composition and layout in the Plt biosynthesis gene cluster among P. proteagens H78 and other four strains. The H78 strain shares the 99% to 100% identity of amino acid sequence of the Plt genes with P. protegens CHA0 and Pf-5, while a relatively low level of identity(52%~88%) with P. aeruginosa M18 and LESB58. The fermentation results show that Plt in P. protegens H78 can accumulate beyond 30μg/m L. Based genomic analysis and antibiotic production, we decide to classify H78 into the kind of P. protegens. There are two distinct evolutionary origins and pathways in the Plt gene cluster. They are respectively the P. protegens type and the P.aeruginosa type. The wild-type strain of H78 can synthesis and secrete the higher yield of Plt and thus has the potential to become a productive and safe strain through further research and engineering.

关 键 词：假单胞菌(P.protegens)H78 藤黄绿菌素 生物合成基因簇 

分 类 号：Q78[生物学—分子生物学] S476[农业科学—农业昆虫与害虫防治]