Singh Manoj Kumar, Kushwaha Chanda, Singh Ramesh Kumar
Microbial Genetics Laboratory, Genetics and Plant Breeding, Institute of Agricultural Sciences, Banaras Hindu University, Varanasi, 221005, India.
Curr Microbiol. 2009 Sep;59(3):240-3. doi: 10.1007/s00284-009-9419-6. Epub 2009 May 30.
Colonization ability of the two endophytic bacteria, isolated from surface sterilized roots of upland cultivated rice viz., Rhizobium sp. and Burkholderia sp., was compared after genetically tagging them with a constitutively expressing green fluorescent protein gene (gfp/gusA). Confocal laser scanning microscopy (CLSM) of gnotobiotically grown seedlings of Narendradhan 97, inoculated with gfp/gusA-tagged endophytes, revealed that both Rhizobium sp. and Burkholderia sp. colonized the intercellular spaces in the root cortex when inoculated separately. Colonization by gfp/gusA-tagged Rhizobium sp. was severely inhibited when co-inoculated with an equal number (10(6) cfu ml(-1)) of wild type Burkholderia sp. Burkholderia sp. was a more aggressive endophytic colonizer of rice than Rhizobium sp. The potential of using gfp/gusA reporter and CLSM as tools in evaluating competitive ability of colonization among endophytes is demonstrated in this study.
从旱稻表面消毒的根部分离出的两种内生细菌,即根瘤菌属(Rhizobium sp.)和伯克霍尔德菌属(Burkholderia sp.),在用组成型表达绿色荧光蛋白基因(gfp/gusA)进行基因标记后,比较了它们的定殖能力。对接种了gfp/gusA标记内生菌的无菌培养的Narendradhan 97幼苗进行共聚焦激光扫描显微镜(CLSM)观察,结果显示,单独接种时,根瘤菌属和伯克霍尔德菌属均定殖于根皮层的细胞间隙。当与等量(10(6) cfu ml(-1))的野生型伯克霍尔德菌属共同接种时,gfp/gusA标记的根瘤菌属的定殖受到严重抑制。伯克霍尔德菌属比根瘤菌属更具侵略性,是水稻内生定殖菌。本研究证明了使用gfp/gusA报告基因和CLSM作为评估内生菌定殖竞争能力工具的潜力。