中国中药杂志

2021, v.46(04) 915-922

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不同生境滇重楼根际解磷菌的筛选与鉴定
Isolation and identification of phosphatolytic bacteria in Paris polyphylla var. yunnanensis

杜慧慧;朱芙蓉;杨敏;郭冬琴;赵顺鑫;李庆天;周浓;
DU Hui-hui;ZHU Fu-rong;YANG Min;GUO Dong-qin;ZHAO Shun-xin;LI Qing-tian;ZHOU Nong;Chongqing Engineering Laboratory for Green Cultivation and Deep Processing of the Three Gorges Reservoir Area′s Medicinal Herbs,College of Biological and Food Engineering,Chongqing Three Gorges University;College of Pharmacy and Chemistry,Dali University;

摘要(Abstract):

二级濒危药用植物滇重楼Paris polyphylla var.yunnanensis野生资源严重匮乏,引种栽培可缓解市场需求,筛选滇重楼根际土壤解磷菌,为开发高效微生物菌肥提供数据支撑。该研究从云南、四川和贵州3个地区10个不同地点的野生和移栽品种的滇重楼根际土壤中分离筛选具有矿化有机磷能力的根际解磷菌,经分离纯化后定性定量分析解磷能力,结合生理生化实验,运用16S rDNA测序分析进行菌株鉴定。从10个不同生境滇重楼根际土壤中筛选41株解有机磷菌株,其中野生品种滇重楼根际土壤中获得21株,移栽品种根际土壤中获得20株。显著性分析发现,41株解有机磷菌株的解磷能力存在明显差异,溶磷量为0.08~67.61 mg·L~(-1),pH 4.27~6.82。其中菌株Y3-5的溶磷量为67.61 mg·L~(-1),增磷量为57.57 mg·L~(-1)。经鉴定41株均为革兰阳性芽孢杆菌。结合生理生化和系统进化树,最终筛选运动芽孢杆菌Y3-5为候选滇重楼根际解磷菌。不同生境滇重楼根际土中解磷菌的分布不同,解磷能力存在差异,解有机磷菌株Y3-5可望作为滇重楼微生物菌肥的候选菌种。
The wild resources of Paris polyphylla var. yunnanensis, a secondary endangered medicinal plant, are severely scarce. Introduction and cultivation can alleviate market demand. To screen phosphatolytic bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis and provide data support for the development of high-efficiency microbial fertilizer, in this study, the dilution plate coating method was used to isolate and screen the phosphorus solubilizing bacteria with the ability of mineralizing organic phosphorus from the rhizosphere soil of wild and transplanted varieties of P. polyphylla var. yunnanensis in 10 different locations in Yunnan, Sichuan and Guizhou. After separation and purification, the phosphatolytic capacity was analyzed by qualitative and quantitative analysis. Combined with physiological and biochemical experiments, the strains were identified using 16 S rDNA sequencing analysis. Forty one strains were selected from the rhizosphere soil of P. polyphylla var. yunnanensis from 10 different habitats. Among them, 21 strains were obtained from the rhizosphere soil of the wild variety P. polyphylla var. yunnanensis and 20 strains were obtained from the rhizosphere soil of the transplanted variety. And significance analysis found that 41 organophosphate solubilizing strains had significant differences in their ability to solubilize phosphorus. The amount of phosphate solubilizing was 0.08-67.61 mg·L~(-1), the pH value was between 4.27 and 6.82. The phosphatolytic amount of strain Y3-5 was 67.61 mg·L~(-1), and the phosphorus increase amount was 57.57 mg·L~(-1). All 41 strains were identified as Gram-positive Bacillus. Combining physiological characteristic and phylogenetic trees, Bacillus mobilis Y3-5 was finally selected as the candidate rhizosphere phosphatolytic bacteria of P. polyphylla var. yunnanensis. The distribution of phosphorus solubilizing bacteria in the rhizosphere soil of P. polyphylla var. yunnanensis was different, and there were significant diffe-rences in phosphorus solubility. Organophosphate-dissolving strain Y3-5 is expected to be a candidate strain of P. polyphylla var. yunnanensis microbial fertilizer.

关键词(KeyWords): 滇重楼;分离鉴定;解磷细菌;系统发育树;微生物菌肥
Paris polyphylla var.yunnanensis;isolation and identification;phosphatolytic bacteria;phylogenetic tree;microbial fertilizer

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81260622);; 重庆市自然科学基金项目(cstc2018jcyjAX0267);; 云南省地方本科高校(部分)基础研究联合专项项目(2019FH001-041)

作者(Author): 杜慧慧;朱芙蓉;杨敏;郭冬琴;赵顺鑫;李庆天;周浓;
DU Hui-hui;ZHU Fu-rong;YANG Min;GUO Dong-qin;ZHAO Shun-xin;LI Qing-tian;ZHOU Nong;Chongqing Engineering Laboratory for Green Cultivation and Deep Processing of the Three Gorges Reservoir Area′s Medicinal Herbs,College of Biological and Food Engineering,Chongqing Three Gorges University;College of Pharmacy and Chemistry,Dali University;

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DOI: 10.19540/j.cnki.cjcmm.20201120.102

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