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.

[1]	方金龙, 王仕禄, 王雨春, 李秋华, 向刚, 李社红, 包宇飞, 刘坤, 郭明伟, 檀迪, 韩祥峰. 怒江和澜沧江上游段河流浮游植物群落特征及其水生态驱动作用 [J]. 生态学杂志, 2025, 44(12): 4052-4061.
[2]	万佳静, 景元书, 吉梦宇, 孙琪娜. DNDC模型在稻田水肥管理中的应用研究进展 [J]. 生态学杂志, 2024, 43(7): 2198-2207.
[3]	杜红春, 朱挺兵, 龚进玲, 胡飞飞, 朱永久, 杨德国. 澜沧江西藏段浮游植物群落结构特征及其与环境因子的关系 [J]. 生态学杂志, 2022, 41(9): 1769-1777.
[4]	左艳洁, 戍国标, 王崇云, 彭明春, 谢映旭, 张秋媛, 李富利. 澜沧江中上游河谷植被的群落数量分类和谱系结构 [J]. 生态学杂志, 2021, 40(9): 2665-2677.
[5]	李睿,张黎,景元书,李攀,任小丽,何洪林,吕妍. 基于FLUXNET的CLM模型总初级生产力模拟评价与误差分析 [J]. 生态学杂志, 2019, 38(9): 2883-2895.
[6]	李亦然,马睿,张永涛,程甜甜,李泽东,牛勇. 基于Gash修正模型模拟侧柏及其混交林的林冠截留过程 [J]. 生态学杂志, 2019, 38(5): 1331-1338.
[7]	张雪蕾,王义成,肖伟华,杨瑞祥,王燕,朱丽姗. 石羊河流域NPP对气候变化的响应 [J]. 生态学杂志, 2018, 37(10): 3110-3118.
[8]	陶正华1,2,赵志琦1**,张东1,3,李晓东1,王宝利1,吴起鑫1,4,张伟1,5,刘丛强1. 西南三江（金沙江、澜沧江和怒江）流域化学风化过程 [J]. 生态学杂志, 2015, 34(8): 2297-2308.
[9]	高婵婵1,彭焕华2,赵传燕1**,刘兴明3. 基于Gash模型的青海云杉林降水截留模拟 [J]. 生态学杂志, 2015, 34(1): 288-294.
[10]	丁城志1,2,陈毅峰1**,姚景龙1,何德奎1. 澜沧江云南吻孔鲃的耳石年轮特征及其年龄鉴定 [J]. 生态学杂志, 2013, 32(11): 3069-3073.
[11]	张昌顺1,谢高地1**,包维楷2,陈龙1，3,裴厦1,范娜1. 地形对澜沧江源区高寒草甸植物丰富度及其分布格局的影响 [J]. 生态学杂志, 2012, 31(11): 2767-2774.
[12]	王海燕;高增祥;邹涛;刘光亮;刘哲. 渤海淡水存留时间分析 [J]. 生态学杂志, 2010, 29(03): 498-503.
[13]	刘世梁, 杨志峰, 崔保山, 甘淑. 道路对景观的影响及其生态风险评价——以澜沧江流域为例 [J]. 生态学杂志, 2005, (8): 897-901.