PhoR/PhoP双因子调控系统在枯草芽孢杆菌NCD-2菌株解磷能力中的功能分析

›› 2009, Vol. 11 ›› Issue (1): 55-61.

PhoR/PhoP双因子调控系统在枯草芽孢杆菌NCD-2菌株解磷能力中的功能分析

李晶1,2,郭庆港2,李社增2,马平2

. 河北大学生命科学学院, 河北保定 |071000； 2. 河北省农林科学院植物保护研究所,
河北省农业有害生物综合防治工程技术研究中心, 河北保定 |071000

收稿日期:2008-11-07 修回日期:2008-12-10 出版日期:2009-02-15 发布日期:2009-07-29
通讯作者: 马平,研究员,主要从事植物病理学和植物病害生物防治。E-mail:pingma88@126.com。郭庆港,博士,助理研究员,主要从事植物病害生物防治。E-mail: gqg77@163.com
作者简介:李晶,硕士研究生,主要从事植物病害生物防治研究工作。E-mail:lishaojing2002@yahoo.com.cn。
基金资助:
国家863计划项目（2006AA10A211）；国家自然科学基金项目（30671401）；河北省农林科学院财政专项（2007055001）资助。

|Function Analysis of PhoR/PhoP Twocomponent Regulatory System for Lecithinsolubilizing Ability in Bacillus subtilis Strain NCD-2

|LI Jing1,2, GUO Qing-gang2, LI She-zeng2, MA Ping2

1. College of Life Science, Hebei University, Hebei Baoding 071000|2. IPM Centre of Hebei Provence,
Institute of Plant Protection, Hebei Academy of Agriculture and Forestry Sciences, Hebei
Baoding 071000, China

Received:2008-11-07 Revised:2008-12-10 Online:2009-02-15 Published:2009-07-29

摘要/Abstract

摘要：

枯草芽孢杆菌（Bacillus subtilis）NCD-2能有效的防治棉花黄萎病,同时还具有降解蛋黄卵磷脂的能力。将含有miniTn10转座子的pHV1249质粒电击转入NCD-2菌株中,51℃高温诱导转座子插入突变,构建了NCD-2菌株的解磷突变子库,筛选到3株丧失解磷能力的突变子。运用染色体步移技术对突变株M216中转座子插入位点基因的侧翼序列进行克隆和序列分析,结果表明丧失解磷能力突变株中转座子插入基因与B.subtilis 168菌株中PhoR基因的相似率达到98%,Southern杂交验证突变株中转座子为单拷贝插入。将PhoR基因克隆到pHY300PLK质粒上构建重组质粒电击转化M216进行功能互补验证,互补菌株部分恢复了解磷能力,表明NCD2菌株中PhoR基因与其降解卵磷脂能力具有相关性。

关键词: 草芽孢杆菌；转座子miniTn10；降解卵磷脂;PhoR基因

Abstract:

Bacillus subtilis strain NCD-2 can effectively control cotton verticillium wilt, it also has lecithin (or phosphatidylchoine)solubilizing ability. Plasmid pHV1249 carrying transposon miniTn10 was transferred into strain NCD-2 by electrotransformation, and the transposon insertional mutagenesis was induced at 51℃. miniTn10 insertion mutants were assayed for the lecithinsolubilizing ability, 3 mutants losing lecithinsolubilizing ability were obtained. The flanking sequence of insertion site was cloned from mutant M216 by chromosome walking technique. Sequence analysis showed that the inserted gene was of 98% identity with PhoR of B.subtilis strain 168. Southern blot hybridization proved that strain NCD-2 was inserted by a singlecopy transposon. Genetic complementation was conducted by cloning PhoR into plasmid pHY300PLK and the recombinant plasmid was transferred into mutant M216. The lecithinsolubilizing ability of mutant M216 was restored. The results suggested that the lecithinsolubilizing ability of NCD-2 was related with PhoR gene.

Key words: Bacillus subtilis, transposon miniTn10, lecithin solubilization, PhoR gene

中图分类号:

李晶1,2,郭庆港2,李社增2,马平2.
PhoR/PhoP双因子调控系统在枯草芽孢杆菌NCD-2菌株解磷能力中的功能分析[J]. , 2009, 11(1): 55-61.

LI Jing1,2, GUO Qing-gang2, LI She-zeng2, MA Ping2. |Function Analysis of PhoR/PhoP Twocomponent Regulatory System for Lecithinsolubilizing Ability in Bacillus subtilis Strain NCD-2[J]. , 2009, 11(1): 55-61.

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