烟草根际土壤中解钾细菌的分离与多样性分析
Diversity and isolation of potassium solubilizing bacteria in tobacco rhizosphere soils
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摘要: 土壤中含有丰富的钾元素, 但主要以缓效态形式存在于钾长石或云母等硅酸盐矿物中, 不能被作物直接吸收利用。解钾微生物能溶解硅酸盐矿物中的钾, 提高土壤中作物可利用钾的含量, 有望缓解我国钾肥短缺的现状。本研究利用选择性培养基, 从烟草根际筛选钾细菌, 基于16S rDNA序列分析烟草根际土壤解钾细菌的多样性, 通过测定解钾细菌的解钾效能及对烟草的促生作用, 筛选有应用潜力的优良解钾细菌菌株。结果表明, 从四川、湖北和山东烟区烟草根际土壤分离获得的27株解钾细菌, 在解钾固体培养上溶钾圈直径为0.11~0.30 cm。16S rDNA序列分析表明, 烟草根际土壤解钾细菌主要包括变形菌门γ亚群(Gammaproteobacteria, 85.18%)、变形菌门α亚群(Alphaproteobacteria, 3.70%)、变形菌门β亚群(Betaproteobacteria, 3.70%)、放线菌门(Actinobacteria, 3.70%)和拟杆菌门(Bacteroidetes, 3.70%), 其中克雷伯菌属(Klebsiella)为优势菌属(66.67%)。27个菌株均有一定的解钾能力, 解钾活性为0.59~4.40 mg·L-1。参试菌株均对烟草有一定的促生作用, 利用解钾细菌菌液处理烟株20 d后, 与对照相比, 株高增加0.97%~38.64%, 最大叶长增加4.40%~31.02%。本研究筛选出的菌株XF11、GM2、JM19和GL7具有较高的解钾活性和促进植物生长的能力, 展现了良好的应用潜力。Abstract: The concentration of soluble K in soils is usually very low which implies that a higher proportion of soil K occurs as insoluble rock, minerals and other deposits. K solubilizing bacteria have been noted to dissolve K from insoluble minerals, increase available K content in soils and stimulate plant growth. Application of K solubilizing bacteria may relieve K fertilizer shortage in China. In this research, K solubilizing bacteria were isolated in selective medium from tobacco rhizosphere soils sampled from tobacco areas in Sichuan, Hubei and Shangdong Provinces, and the diversity of the bacteria isolates analyzed using the 16S rDNA sequence. 27 K solubilizing bacteria strains were isolated and all the isolated bacteria strains solubilized powdered feldspar in solid and liquid media. The range of the zone of solubilization diameter formed by the strains was 0.11~0.30 cm on the agar medium. All the strains were able to release K (0.59~4.40 mg·L-1 K of the fermentation broth) and plant-promoting effects (plant height and longest leaf length respectively increased by 0.97%~38.64% and 4.40%~31.02%). Strains XF11, GM2, JM19 and GL7 were high-efficient strains for K release and plant growth promotion and had high application potential. The strains were identified through comparisons of 16S ribosomal DNA sequences. There were 85.18% Gammaproteo-bacteria, 3.70% Alphaproteo-bacteria, 3.70% Betaproteo-bacteria, 3.70% Actins-bacteria and 3.70% Bacteroidetes. Among the isolated strains, the dominant bacteria were the strains for the genera Klebsiella bacteria in tobacco rhizosphere soil. Further studies were necessary into the effects of the bacterial strains on mobilizing K-bearing minerals under field conditions.