小白菜根际高效解磷菌的分离鉴定与培养条件优化

    Isolation and Identification of the High Efficient Phosphate-solubilizing Bacteria from the Rhizosphere of Brassica chinensis L. and the Optimization of Culture Conditions

    • 摘要: 为评价小白菜Brassica chinensis L.根际微生物分解有机磷的能力,明确其优势解磷菌的分类学地位,进一步优化培养条件,为开发微生物肥料提供基础。采用蒙金娜有机磷平板法筛选小白菜根际有机磷分解菌,利用钼锑抗比色法分析菌株解磷能力;进一步观察优势解磷菌的菌落形态,并通过16S DNA序列同源性比对和邻近法构建系统发育树,确定优势菌株FG5的分类学地位;最后,采用单因素和正交试验进一步探讨菌株FG5的最适培养条件。结果表明:小白菜根际共分离筛选出解有机磷菌3株,其中菌株FG5解磷效果最佳,接种菌株FG5培养3 d后,培养基中游离磷质量浓度高达165.20 ug·mL-1。FG5菌落圆形,表面干燥,微微隆起,边缘波状的米白色,16S DNA序列同源性比对结果表明其为特基拉芽孢杆菌。单因素和正交试验优化结果显示菌株FG5最佳培养条件为:甘露醇为碳源,尿素为氮源,培养温度27℃,转速180 r·min-1,接种量1.5 %(V/V)。在该条件下FG5的解磷量达到307.25 ug·mL-1,相较于培养条件优化前,解磷能力提高85.99%。菌株FG5有机磷分解能力强,具有良好的微生物肥料应用潜力。

       

      Abstract: In order to evaluate the ability of microorganism in the rhizosphere of Brassica chinensis L. to decompose the organic phosphorus, the taxonomic status of its dominant phosphate-solubilizing bacteria was clarified, and the culture conditions were further optimized, thus to provide the basis for the development of microbial fertilizers. The organic phosphorus-decomposing bacteria in the rhizosphere of Brassica chinensis L. were screened by using the Mengjinna organic phosphorus plate method, and the phosphate-solubilizing ability of the strains was analyzed through the molybdenum antimony resistance colorimetric method. Then, the colonial morphology of the dominant phosphate-solubilizing bacteria was further observed. And the phylogenetic tree was constructed through the homology comparison of 16S DNA sequence and nearest neighbor method to determine the taxonomic status of the dominant strain FG5. Last, the optimum culture conditions of strain FG5 were further explored by using the single factor and orthogonal experiments. The results showed that: three strains of organic phosphate-solubilizing bacteria were isolated and screened from the rhizosphere of Brassica chinensis L., among which the strain FG5 had the best phosphate-solubilizing effect. After the inoculation with strain FG5 and culture for 3 days, the mass concentration of free phosphorus in the medium was as high as 165.20 ug·mL-1. The colony of FG5 was circular with a dry surface and slight protrusions, and the edges were wavy off white. The homology comparison results of the 16S DNA sequence indicated that the strain FG5 was Bacillus tequilensis. The results of single factor and orthogonal experiments showed that the optimum culture conditions for the strain FG5 were as follows: mannitol as carbon source, urea as nitrogen source, culture temperature of 27℃, rotational speed of 180 r·min-1, inoculation amount of 1.5% (V/V). Under this condition, the phosphate-solubilizing ability of FG5 reached 307.25 ug·mL-1, which was 85.99% higher than that before the optimization of culture conditions. The strain FG5 had a strong ability to decompose the organic phosphorus and had good potential for the application of microbial fertilizers.

       

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