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草业学报 ›› 2011, Vol. 20 ›› Issue (4): 244-251.

• 研究论文 • 上一篇    下一篇

四株胡萝卜软腐欧文氏杆菌胡萝卜亚种新菌株的分离鉴定

赵玲1,2,3,4,柴兆祥1,李金花1,2,3,4*,王蒂2*   

  1. 1.甘肃农业大学草业学院,甘肃 兰州 730070;
    2.甘肃省作物遗传改良与种质创新重点实验室,甘肃 兰州 730070;
    3.草业生态系统教育部重点实验室,甘肃 兰州 730070;
    4.中-美草地畜牧业可持续发展研究中心,甘肃 兰州 730070
  • 收稿日期:2010-03-18 出版日期:2011-04-25 发布日期:2011-08-20
  • 通讯作者: E-mail:li_jinhua@yeah.net,wangd@gsau.edu.cn
  • 作者简介:赵玲(1985-),女,黑龙江齐齐哈尔人,助教,硕士。E-mail:751470508@qq.com
  • 基金资助:
    甘肃省农业生物技术研究与应用开发项目(GNSW-2006-01),现代农业产业技术体系建设专项资金(CARS-10-P18)和国家科技支撑计划项目(2006BAD21B05)资助。

Isolation and identification of four new Erwinia carotovora subsp. carotovora strains

ZHAO Ling1,2,3,4,CHAI Zhao-xiang1,LI Jin-hua1,2,3,4,WANG Di2   

  1. 1.College of Grassland Sciences, Gansu Agricultural University, Lanzhou 730070, China;
    2.Gansu Key Lab of Crop Improvement and Germplasm Enhancement, Lanzhou 730070, China;
    3.Key Laboratory of Grassland Ecosystem, Ministry of Education, Lanzhou 730070, China;
    4.Sino-U.S. Centers for Grazingland Ecosystem Sustainability, Lanzhou 730070, China
  • Received:2010-03-18 Online:2011-04-25 Published:2011-08-20

摘要: 采用棋盘式取样法,从正茬、重茬、连作2年和连作3年的马铃薯播前、现蕾期、成熟期和收获期4个不同阶段采集马铃薯根围土样。以稀释平板法,用选择性培养基CVP分离软腐欧文氏杆菌,共得到42个菌株。在形态学观察和生理生化特性测定的过程中发现编号为GAUP-b410、GAUP-b492、GAUP-b416和GAUP-d34等4个菌株的主要性状一致,且与其他菌株的性状不同。致病性测定结果显示,这4个菌株均能引起马铃薯薯片腐烂。对其中的2个菌株进行了16S rDNA 序列分析,其PCR产物的序列与GenBank 上登记的18个分离自中国大白菜上的Erwinia carotovora subsp. carotovora菌株的亲缘关系最近,同源性均达99%。利用DNAStar软件绘制系统发育树状图,菌株GAUP-b410亦与这18个E. carotovora subsp. carotovora菌株位于系统发育树的同一分支,聚为一类;菌株GAUP-b492与其中的16个E. carotovora subsp. carotovora菌株位于系统发育树的同一分支,聚为一类;结合生理生化特性和16S rDNA序列分析,确定这4个菌株为胡萝卜软腐欧文氏杆菌胡萝卜亚种(E. carotovora subsp. carotovora)新菌株。

Abstract: Rhizosphere soils of potato (Solanum tuberosum) were sampled via a chess-board pattern from fields of potato grown for one year, continuously for 2 years, 3 years and 4 years at 4 growing stages (before planting, squaring stage, maturity stage and harvest stage). The soils were isolated on CVP medium via the dilution method for soft rot Erwinia and 42 strains were obtained. During observations of morphology and tests of their physiological and biochemical characteristics, it was found that 4 of the 42 strains shared the same characteristics but differed from the other strains. The 4 strains were GAUP-b410, GAUP-b492, GAUP-b416 and GAUP-d34. The pathogenicity tests indicated that these 4 strains were artificially pathogenetic to cut potato pieces. GAUP-b410 and GAUP-b492 were taken at random for analysis of 16S rDNA sequence, which showed 99% sequence identity with 18 E. carotovora subsp.carotovora strains from Chinese cabbage in China after aligning data in GenBank. Microsoft DNAStar was used to draw the phylogenetic tree of strain GAUP-b410 and GAUP-b492. The tree showed that strain GAUP-b410 was in the same cluster as the 18 Chinese strains, while strain GAUP-b492 was in the same cluster as the 16 of 18 Chinese strains. Therefore, based on the physiological and biochemical characteristics of the 4 strains and the result of 16S rDNA sequence analysis of the 2 strains, the 4 strains were identified as the E. carotovora subsp.carotovora, which were 4 new E. carotovora subsp.carotovora strains.

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