解磷细菌 K3 的 GFP 标记及其解磷能力检测
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国家自然科学基金项目(30771258)和国家863项目(2006AA10Z416)资助


Labeling of phosphate-solubilizing bacteria K3 with GFP and its phosphate solulilization ability
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    摘要:

    本文运用构建成功的含有假单胞菌属自身启动子及绿色荧光蛋白(GFP)的质粒 pTRGFP 电转至假单胞菌解磷细菌 K3 中,通过激光共聚焦显微镜及质粒检测,获得了发光稳定的标记菌株 K3GFP。7 天液体摇瓶试验中,标记菌株 K3GFP的可溶性 P 含量在第 4 天达到最高 774 μg/ml,而出发菌株 K3 在第 3 天时已经达到了最大含量 780 μg/ml,说明 pTRGFP 质粒的转入对 K3 菌株解 P 能力有一定的影响。标记菌株 K3GFP 施入自然土壤 10 天后数量维持在 5.47×106 CFU/g ~ 2.40×106 CFU/g ,35 天后降到 5.0×103 CFU/g ,说明解磷细菌 K3GFP 可以在自然土壤中定殖。本试验为研究解磷细菌在根际及土壤中的生长动态等行为特征奠定了基础。

    Abstract:

    In this study, the plasmid pTRGFP with Pseudomonas promoter and green fluorescent protein (GFP) segment was transformed into strain K3 by electroporation, and the conjugant K3GFP with steady fluorescence was acquired. Both fluorescent microscope and plasmid analysis were used to verify the success of the electroporation. The available P concentration reached the peak (770 μg/ml) at the 4th day for the labeling strain K3GFP while the wild strain achieved the most (780 μg/ml) at the 3rd day. The result demonstrated that phosphate-solubilizing ability of the wild strain was affected after the electroporation. The total population of K3GFP strain was 5.47×106 - 2.40×106 CFU/g soil from 10 to 28 days, and decreased to about 5.00×103 CFU/g soil after 35 days in a natural soil. The result indicated the K3GFP had strong colonization ability in soil. The K3GFP will provide a foundation to the further research on phosphate-solubilizing bacteria.

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李晓婷,董彩霞,杨兴明,钟增涛,沈其荣,徐阳春.解磷细菌 K3 的 GFP 标记及其解磷能力检测[J].土壤,2010,42(4):548-553. LI Xiao-ting, DONG Cai-xia, YANG Xing-ming, ZHONG Zeng-tao, SHEN Qi-rong, XU Yang-chun. Labeling of phosphate-solubilizing bacteria K3 with GFP and its phosphate solulilization ability[J]. Soils,2010,42(4):548-553

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