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1.4×104cfu/g和7.6×104cfu/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9.1×104cfu/g和3.17×105cfu/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。
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× 10^4cfu/g and 7.6×10^4cfu/g, respectively. The colonization was very stably. The number of cells on celery root surface and soil around the roots were still 9.1×10^4cfu/g and 3.17×10^5cfu/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.
出处
《中国农学通报》
CSCD
2014年第9期237-241,共5页
Chinese Agricultural Science Bulletin
基金
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)
