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阿尔山国家森林公园自然保护区杨树根际土壤固氮菌多样性及特性研究
牛艳芳1,2, 闫 伟1, 陈立红,等1
1.内蒙古农业大学 林学院;2.内蒙古师范大学 生命科学与技术学院
摘要:
【目的】从杨树根际土壤筛选固氮菌,并研究其生长特性。【方法】在内蒙古的阿尔山国家森林公园自然保护区,采集杨树根际土壤,采用阿须贝无氮培养基分离纯化杨树根际土壤固氮菌。对分离纯化的固氮菌进行形态和生理生化鉴定;用细菌通用引物27f/1492r 对固氮菌16S rDNA序列进行扩增并测序,应用DNAMAN6.0软件进行序列校正及多序列比对,使用MAGE 4中的邻接法(NJ)构建系统发育树。对分离所获固氮菌的生长特性和解磷能力进行测定。【结果】从阿尔山杨树根际土壤中共分离出XY1、XY2、XY5、XY7、XY8、XY14、XY15、XY9等8株固氮菌,菌落形态、生理生化特性鉴定及系统发育树结果显示,以上8株固氮菌分别属于柄杆菌属(Caulobacter)、叶杆菌属(Phyllobacterium)、红球菌属(Rhodococcus)、假单胞菌属(Pseudomonas)、根瘤菌属(Rhizobium)、类芽孢杆菌属(Paenibacillus)、沙雷氏菌属(Serratia)、芽孢杆菌属(Bacillus)。不同固氮菌生长的最适pH和温度有差异。分离得到的固氮菌中,只有XY2和XY7具有解磷活性。【结论】阿尔山自然保护区杨树根际土壤固氮菌资源丰富,其生物学特性有明显差异。
关键词:  阿尔山  杨树  根际  固氮菌  16S rDNA
DOI:
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基金项目:国家自然科学基金项目(31260173)
Diversity and characterization of nitrogen-fixing bacteria in Populus L.rhizosphere in Arxan National Forest Park
NIU Yanfang,YAN Wei,CHEN Lihong,et al
Abstract:
【Objective】This paper screened nitrogen fixing bacteria from Populus L.rhizosphere and studied their growth characteristics.【Method】In Arxan National Forest Park of Inner Mongolia,nitrogen-fixing bacteria were isolated from soil of Populus L.rhizosphere and purified by Ashby’s medium.Then morphological and physiology biochemical identification was conducted and the 16S rDNA was amplified with primers 27f/1492r and sequenced.Neighbor-joining tree was also built with DNAMAN6.0 and MEGA 4 and the growth and capacity of phosphate solubilization were measured.【Result】Eight nitrogen-fixing bacterium including XY1,XY2,XY5,XY7,XY8,XY14,XY15 and XY9 were isolated in soil of Populus L.rhizosphere.They belonged to Caulobacter,Phyllobacterium,Rhodococcus,Pseudomonas,Rhizobium,Paenibacillus,Serratia and Bacillus,and had different optimum temperatures and pH values.Only XY2 and XY7 had phosphate solubilization capacity.【Conclusion】Nitrogen-fixing bacteria in Populus L. rhizosphere in Arxan National Forest Park had rich diversity,and there were significant differences in their biological characteristics.
Key words:  Arxan  Populus L  rhizosphere  nitrogen-fixing bacteria  16S rDNA