GFP标记内生枯草芽孢杆菌Y10及其在白菜体内的定殖

生态学杂志

GFP标记内生枯草芽孢杆菌Y10及其在白菜体内的定殖

杜芳¹,何鹏飞¹,吴毅歆^2,3,陈卓君¹,杨静¹,何月秋^2,3**

(¹云南农业大学植物保护学院，昆明 650201； ²微生物菌种筛选与应用国家地方联合工程研究中心，昆明 650207； ³云南农业大学农学与生物技术学院，昆明 650201)

出版日期:2015-07-10 发布日期:2015-07-10

Colonization of GFPtagged endophytic Bacillus subtilis Y10 in Chinese cabbage.

DU Fang¹, HE Peng-fei¹, WU Yi-xi^2,3, CHEN Zhuo-jun¹, YANG Jing¹, HE Yue-qiu^2,3**

(¹Faculty of Plant Protection, Yunnan Agricultural University, Kunming 650201, China; ^2National and Local Joint Engineering Research Center for Screening and Application of Microbial Strains, Kunming 650207, China; ³Faculty of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming 650201, China)

Online:2015-07-10 Published:2015-07-10

摘要/Abstract

摘要：

枯草芽孢杆菌Y10是具有防治根肿病的白菜内生菌株。为研究其在白菜体内的定殖规律，将不同来源的含绿色荧光蛋白（GFP）质粒导入Y10进行荧光标记，测定了荧光标记菌的质粒稳定性、生长曲线及室内平板拮抗活性。结果表明，仅有pHT01P43GFPmut3a能在Y10中稳定地遗传，且对Y10的生长和病原真菌拮抗活性无不利影响。盆栽防效试验结果显示，在不同的栽培基质中，Y10的标记菌株与野生型对白菜根肿病防效无显著差异。稀释涂板回收及共聚焦激光扫描显微观察都证实，接种9 d后在白菜的根、茎及叶部组织均有标记菌株的定殖，且在44 d内仍保持10³～10⁴ cfu·g^-1组织。这些定殖结果表明，Y10在白菜体内具有良好的定殖能力，且这种特性可能与其防治根肿病有关。

关键词: 敏感性分析, 变异度, Hurst系数, 澜沧江, Bartels统计量

Abstract:

Endophytic Bacillus subtilis Y10 isolated from Chinese cabbage (Brassica rapa) can control effectively cruciferous vegetable clubroot disease. To understand its clubrootcontrol mechanisms, GFP plasmids were transformed into Y10 cells for fluorescencetagging by natural transformation. The plasmid stability, growth curve and plate antagonism bioassay of GFPtagged strains were determined, and the results showed only plasmid pHT01P43GFPmut3a could be maintained stably and its introduction had no significant negative influence on Y10 growth and antagonistic performance. Pot bioassay experiment with different cultivation matrixes indicated that there was no significant difference in clubrootcontrol efficiency between Y10P43GFPmut3a and its wild type. Recovery and observation by confocal laser scanning microscope confirmed that, 9 days after inoculation, tagged strain could colonize the root, stem and leaf of Chinese cabbage, and the colonization density still kept at 10³-10⁴ cfu·g^-1 tissues in 44 days after inoculation. All the results suggest that the clubrootcontrol of Y10 be intimately associated with efficient colonization in Chinese cabbage host.

Key words: sensitivity analysis, Hurst coefficient, Bartels statistics, variability, Lancang River.

杜芳1,何鹏飞1,吴毅歆2,3,陈卓君1,杨静1,何月秋2,3**. GFP标记内生枯草芽孢杆菌Y10及其在白菜体内的定殖[J]. 生态学杂志.

DU Fang1, HE Peng-fei1, WU Yi-xi2,3, CHEN Zhuo-jun1, YANG Jing1, HE Yue-qiu2,3**. Colonization of GFPtagged endophytic Bacillus subtilis Y10 in Chinese cabbage.[J]. cje.

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