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中国农学通报 ›› 2014, Vol. 30 ›› Issue (9): 237-241.doi: 10.11924/j.issn.1000-6850.2013-1713

所属专题: 生物技术

• 生物技术科学 • 上一篇    下一篇

绿色荧光蛋白标记的枯草芽胞杆菌R31在西芹根际定殖研究

陈燕红 黎永坚 喻国辉 陈远凤   

  • 收稿日期:2013-06-21 修回日期:2013-08-26 出版日期:2014-03-25 发布日期:2014-03-25
  • 基金资助:
    珠海市科技计划项目“尖孢镰刀菌引起的西芹香蕉病害的生物防治技术研究”(PC20081078);广东省农业厅农业科技推广专项“枯草芽胞杆菌R31防治香蕉枯萎病的示范推广”(粤财农[2010]540号)。

Colonization of Green Fluorescent Protein Gene Tagged Bacillus subtilis R31 in Celery Rhizosphere

  • Received:2013-06-21 Revised:2013-08-26 Online:2014-03-25 Published:2014-03-25

摘要: 为了更直观地了解植物内生枯草芽胞杆菌R31在西芹根际定殖情况,评估其定殖能力,为西芹黄萎病生物防治提供依据。通过原生质体转化法将携带绿色荧光蛋白基因的质粒pHT315-gfp导入枯草芽胞杆菌R31细胞中,对菌株进行gfp标记,然后利用gfp标记菌株灌根处理西芹苗,研究其在西芹根际的定殖形式和能力。结果获得了携带pHT315-gfp质粒的R31-gfp菌株,该质粒可以在R31细胞中稳定表达,标记菌R31-gfp在蓝光激发下可以稳定发出强的绿色荧光。利用标记菌接种西芹幼苗后,可以在根系表皮检测到绿色荧光膜。对其定殖量的检测发现,接种3天后标记菌在根表和根际土中稳定定殖,定殖量分别达到1.4×104 cfu/g和7.6×104 cfu/g,并长期以该水平稳定定殖,到第65天仍可在根表和根际土检测到9.1×104 cfu/g和3.17×105 cfu/g的R31-gfp。由此得出,R31可以在西芹根表形成菌膜,并在根际长期稳定定殖。

关键词: 香吃味, 香吃味

Abstract: In order to study the colonization of Bacillus subtilis R31 in celery rhizosphere, to assess its colonization ability and provide basis to celery wilt biocontrol. Plasmid pHT315-gfp was introduced into cells of Bacillus subtilis R31 by protoplasts conversion and gfp- tagged R31 was acquired, gfp- tagged R31 was irrigated around the celery root to study its colonization style and ability. Results showed the recombined plasmid was stable in gfp-tagged R31 cells. The R31-gfp cells could emit bright green fluorescence under blue light stably. Green fluorescent film on the surface of the celery root inoculated R31-gfp could be observed under fluorescent microscopy. It was found that R31-gfp cells could colonize in celery rhizosphere quickly. Three days after inoculation, cells on the surface of celery roots and in the soil around the roots were about 1.4×104 cfu/g and 7.6×104 cfu/g, respectively. The colonization was very stably. The number of cells on celery root surface and soil around the roots were still 9.1×104 cfu/g and 3.17×105 cfu/g at the 65 days after inoculation separately. Conclusion: Bacillus subtilis R31 can form biofilm on the surface of the celery roots, and can colonize stably for a long-term in celery rhizosphere.