绿色荧光蛋白标记枯草芽孢杆菌Y13UV在油茶体内的定殖

doi:10.11707/j.1001-7488.20170712

林业科学 ›› 2017, Vol. 53 ›› Issue (7): 111-117.doi: 10.11707/j.1001-7488.20170712

绿色荧光蛋白标记枯草芽孢杆菌Y13^UV在油茶体内的定殖

金勤¹, 朱丹雪¹, 周国英¹, 李河¹, 何苑皞¹, 张茜^1,2

1. 森林有害生物防控湖南省重点实验室经济林培育与保护教育部重点实验室中南林业科技大学长沙 410004;
2. 湖南汽车工程职业学院株洲 412001

收稿日期:2016-04-08 修回日期:2016-12-26 出版日期:2017-07-25 发布日期:2017-08-23
通讯作者: 周国英
基金资助:
"十二五"农村领域国家科技计划课题（2012BAD19B0803）；湖南省研究生科技创新基金项目（CX2015B294）；中南林业科技大学研究生科技创新基金项目（CX2015B15）。

Colonization of GFP-Tagged Bacillus subtilis Y13^UV in Camellia oleifera

Jin Qin¹, Zhu Danxue¹, Zhou Guoying¹, Li He¹, He Yuanhao¹, Zhang Qian^1,2

1. Hunan Provincal Key Laboratoty for Control of Forest Disease and Pests Key Laboratory for Non-Wood Forest Cultivation and Conservation of Ministry of Education Central South University of Forest and Technology Changsha 410004;
2. Hunan Automotive Engineering Vocational College Zhuzhou 412001

Received:2016-04-08 Revised:2016-12-26 Online:2017-07-25 Published:2017-08-23

摘要/Abstract

摘要： [目的]枯草芽孢杆菌Y13^UV对油茶炭疽病具有良好的防治效果，研究其在油茶体内的定殖动态，为油茶炭疽病的防治提供依据。[方法]通过原生质体转化法引入绿色荧光蛋白质粒，构建荧光标记菌株。通过喷叶、灌根、喷叶灌根结合处理多种接种方式及重复接种，测定菌株在油茶体内不同组织部位的定殖数量，分析其定殖规律及能力。[结果]标记菌株可以在油茶根、茎和叶内定殖，单次接种当天，根内回收的标记菌株数量为1.07×10⁵ cfu·g^-1；喷叶灌根结合处理7天后，油茶根、茎和叶内标记菌株的数量分别为8.70×10²、5.00×10²和7.30×10² cfu·g^-1，均高于喷叶、灌根单独处理。重复接种时，油茶根茎叶内标记菌株的定殖量在接种3~5天内达到高峰，然后呈现稳定趋势，20天时定殖量开始大幅下降，第30天喷叶灌根处理的油茶根内的定殖量仅为5.30×10² cfu·g^-1。荧光标记菌株生长较好，稳定表达，对炭疽病菌具有良好的抑制效果。[结论]枯草芽孢杆菌Y13^UV能通过喷叶灌根方式接种在油茶体内定殖并传导，有较好的定殖能力。

关键词: 绿色荧光蛋白, 枯草芽孢杆菌, 油茶炭疽病, 定殖

Abstract: [Objective] Bacillus subtilis Y13^UV can control Colletotrichum gloeosporioides effectively. Studying its colonization dynamics could provide a scientific basis for controlling C. gloeosporioides.[Method] The GFP plasmid was introduced into cells by protoplasts conversion and to acquire GFP-tagged Y13^UV. Camellia oleifera plants were inoculated with the GFP-tagged Y13^UV with different methods including foliar spray, root irrigation, foliar spray plus root irrigation, and single or multiple inoculations. The population numbers of GFP-tagged Y13^UV in different tissues of Camellia oleifera were quantified after inoculations to test the colonization ability of the GFP-tagged strain.[Result] GFP-tagged Y13^UV could colonize in root, stem and leaf tissues of Camellia oleifera. In the same day after single inoculation, the population numbers of the marked strain in root were 1.07×10⁵ cfu·g^-1. Seven days after inoculation by foliar spray plus root irrigation, the population of marked strain in root, stem and leaf tissues of Camellia oleifera were 8.70×102, 5.00×102 and 7.30×10² cfu·g^-1, respectively. And the population numbers were higher than inoculation by foliar spray or root irrigation alone. With multiple inoculations, the population numbers of the tagged strain reached their peaks about 3-5 days after inoculation, the populations kept stable until they dropped dramatically 20 days after inoculation,. The population number of 30th day in root tissue of Camellia oleifera was 5.30×10² cfu·g^-1 when inoculated by foliar spray plus root irrigation. The results showed that GFP-tagged Y13^UV grew better, expressed stably and still had good inhibition to C. gloeosporioides.[Conclusion] After inoculated by foliar spray or root irrigation, GFP-tagged Y13^UV could colonize and transfer in Camellia oleifera, and displayed good colonization ability.

Key words: green fluorescent protein(GFP), Bacillus subtilis, Colletotrichum gloeosporioides, colonization

中图分类号:

S763.13

金勤, 朱丹雪, 周国英, 李河, 何苑皞, 张茜. 绿色荧光蛋白标记枯草芽孢杆菌Y13^UV在油茶体内的定殖[J]. 林业科学, 2017, 53(7): 111-117.

Jin Qin, Zhu Danxue, Zhou Guoying, Li He, He Yuanhao, Zhang Qian. Colonization of GFP-Tagged Bacillus subtilis Y13^UV in Camellia oleifera[J]. Scientia Silvae Sinicae, 2017, 53(7): 111-117.

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绿色荧光蛋白标记枯草芽孢杆菌Y13^UV在油茶体内的定殖

Colonization of GFP-Tagged Bacillus subtilis Y13^UV in Camellia oleifera

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绿色荧光蛋白标记枯草芽孢杆菌Y13UV在油茶体内的定殖

Colonization of GFP-Tagged Bacillus subtilis Y13UV in Camellia oleifera

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绿色荧光蛋白标记枯草芽孢杆菌Y13^UV在油茶体内的定殖

Colonization of GFP-Tagged Bacillus subtilis Y13^UV in Camellia oleifera