Isolation and characterization of a Sinorhizobium fredii mutant that cannot utilize proline as the sole carbon and nitrogen source

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• 作者：Huang Sheng (1) (2) (3)
  Bai Xueliang (1) (2) (3)
  Ma Qingsheng (3)
  Tang Xianlai (3)
  Wu Bo (1) (2) (3)
  
• 关键词：Sinorhizobium fredii ; proline catabolism ; nodulation competitiveness
• 刊名：Chinese Science Bulletin
• 出版年：2004
• 出版时间：November 2004
• 年：2004
• 卷：49
• 期：21
• 页码：2262-2265
• 全文大小：533KB
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• 作者单位：Huang Sheng (1) (2) (3)
  Bai Xueliang (1) (2) (3)
  Ma Qingsheng (3)
  Tang Xianlai (3)
  Wu Bo (1) (2) (3)
  
  1. Key Laboratory of Microbial and Plant Genetic Engineering of Ministry of Education, College of Life Science and Biotechnology, Guangxi University, 530004, Nanning, China
  2. Guangxi Key Laboratory of Subtropical Bioresources Conservation and Utilization; College of Life Science and Biotechnology, Guangxi University, 530004, Nanning, China
  3. College of Life Science and Technology, Guangxi University, 530005, Nanning, China
  
• ISSN：1861-9541

文摘

Sinorhizobium fredii strain HN01 can use proline as the sole carbon and nitrogen source. A mutant strain GXHN100 unable to catabolize proline was screened from 6000 Tn5gusA5 random insertional mutants of S.fredii strain HN01. Sequencing analysis showed that an open reading frame, named pmrA (proline metabolic relative), was inserted by the Tn5gusA5. A positive clone, named pGXHN100 which containing 3.3kb foreign DNA fragment of S. fredii strain HN01, was isolated from a partial gene library of S. fredii HN01 by colony in situ hybridization. Sequence analysis showed that pGXHN100 contained the entire pmrA gene. The 3.3kb DNA fragment of pGXHN100 was cloned into a broad-host-range cosmid vector pLAFR3 to form plasmid pGXHN200 which was subsequently introduced into GXHN100 to form a complemented strain GXHN200. Plant test showed that GXHN100 was effective and no obvious changes in nitrogenase activity comparing with parental strain. But GXHN100 nodulated 2 days later on soybean and its nodulation efficiency and competitiveness were decreased. The complemented strain GXHN200 restored the nodulation efficiency and competitiveness of GXHN100 to the wild type.