薛松 汪军 王国芬 杨腊英 丁兆建 周游 黄俊生

摘 要 采用改良的海藻糖电转化法获得GFP标记菌株X5-GFP、BQA2-GFP，显微镜观察标记菌株在土壤和水培中的番茄根际的定殖情况。结果表明：标记菌株单独或与青枯菌先后加入进行处理，番茄根际标记菌株的种群数量变化趋势都是一致的。在土壤番茄根际中，X5-GFP单独处理后当天的菌量为1.05×106 CFU/g，随之菌量逐渐下降；处理30 d后，为1.50×103 CFU/g；而BQA2-GFP单独处理后当天菌量为1.68×106 CFU/g，随之菌量也逐渐下降；处理30 d后，为2.50×103 CFU/g；在水培番茄根际中，X5-GFP单独处理后当天的菌量为2.70×105 CFU/g，随之菌量逐渐下降；处理30 d后，为1.00×103 CFU/g；而BQA2-GFP单独处理后当天菌量为3.60×105 CFU/g，随之菌量同样逐渐下降；处理30 d后，为1.50×103 CFU/g。这表明解淀粉芽胞杆菌X5、BQA2均在番茄根际有较强的定殖能力。

关键词 解淀粉芽胞杆菌；绿色荧光蛋白；定殖

中图分类号 S432 文献标识码 A

Abstract In this study， green fluorescent protein tagged strains X5 and BQA2 were obtained by improvement in electroporation transformation efficiency of gene for Bacillus amyloliquefaciens with trehalose. X5-GFP and BQA2-GFP colonization in the rhizosphere of tomato were observed by microscope. The experimental results indicated that changing tendency of population quantity in marking strains of tomato rhizosphere was consistent after dealing with individual marking strains or adding Ralstonia solanacearum successively. In the soil， the population of X5-GFP on the roots of tomato reached 1.05×106 CFU/g after inoculating X5-GFP alone， and then decreased 30 days later， eventually remained at 1.50×103 CFU/g. The population of BQA2-GFP on the roots of tomato reached 1.68×106 CFU/g after inoculating BQA2-GFP alone， and then decreased 30 days later， eventually remained at 2.50×103 CFU/g. In Hydroponics， the population of X5-GFP on the roots of tomato reached 2.70×105 CFU/g after inoculating X5-GFP alone， and then decreased 30 days later， eventually remained at 1.00×103 CFU/g. The population of BQA2-GFP on the roots of tomato reached 3.60×105 CFU/g after inoculating BQA2-GFP alone， and then decreased 30 days later， eventually remained at 1.50×103 CFU/g. The results showed that B. amyloliquefaciens X5 and B. amyloliquefaciens BQA2 displayed good colonization ability in the rhizosphere of tomato.

Key words Bacillus amyloliquefaciens； green fluorescent protein； colonization

doi 10.3969/j.issn.1000-2561.2017.03.026

由于人们对粮食质量安全要求的提高，绿色环保粮食已经受到人们的青睐。在生产绿色环保粮食过程中，生物防治由于其環保、低毒等优点受到了广泛的关注[1]。据报道，生防菌在植物上的定殖能力和病害防治效果紧密相关[2-3]，有关研究也成为目前生防菌研究领域的热点之一[4-5]。很多标记方法被开发用于标记目标微生物，使其与其它微生物区别开来，以阐明生防菌的定殖能力。抗生素标记作为最早使用的标记方法，主要有硫酸链霉素、卡那霉素、四环素、氨苄青霉素等标记法[6-7]，但抗生素标记法不能区别出对标记抗生素有同样抗性的其它微生物。随着标记基因……