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为研究青香茅(Cymbopogon caesius)和五节芒(Miscanthus floridulus)内生固氮菌的多样性,基于两种无氮培养基结合乙炔还原法从这两种牧草中分别分离到30株和36株菌.采用SDS-PAGE(Dodecyl sulfate sodium saltpolyacrylamide gel electrophoresis)全细胞蛋白电泳和IS-PCR(Insertion sequence-based PCR)指纹图谱将这66株内生固氮菌聚类为10个类群(I-X,每群3株以上,共61个聚群菌株)和3个未归类小组(共5个未聚群菌株).从10个聚类群和3个未归类小组中各选1株菌作为代表进行16S rDNA序列分析,发现它们与洋葱伯克霍尔德菌(Burkholderia cepacia)、阴沟肠杆菌(Enterobacter cloacae)、固氮植物菌(Phytobacter diazotrophicus)、阿氏肠杆菌(Enterobacter asburiae)、织片草螺菌(Herbaspirillum seropedicae)、阪崎克洛诺菌(Cronobacter sakazakii)、水稻肠杆菌(Enterobacter oryzae)、肺炎克雷伯氏菌(Klebsiella pneumoniae)、溶解肠杆菌(Enterobacter dissolvens)、无乳固氮螺螺(Azospirillum amazonense)、都柏林克罗诺杆菌(Cronobacter dublinensis)等的相似性分别达97%以上,表明所分离内生固氮菌多样性高.进一步对它们进行生理生化测定,结果表明有些菌株具泌氨能力、产吲哚、解磷、解钾效果,在农业生产上具有一定的应用潜力.
In order to study the diversity of endophytic diazotrophs in Cymbopogon caesius and Miscanthus floridulus, 30 and 36 isolates were isolated from these two forages based on two nitrogen-free medium combined with acetylene reduction The 66 strains of endophytic diazotrophs were clustered into 10 groups (IX, each group) using SDS-PAGE (Dodecyl sulfate sodium salt polyacrylamide gel electrophoresis) whole cell protein electrophoresis and IS-PCR (Insertion sequence-based PCR) Three or more, a total of 61 cluster strains) and three unclassified groups (a total of five non-clustered strains) from 10 clusters and 3 unclassified groups each selected one strain as a representative 16S rDNA sequence analysis, they were found to bind to Burkholderia cepacia, Enterobacter cloacae, Phytobacter diazotrophicus, Enterobacter asburiae, Herbaspirillum seropedicae, Cronobacter sakazakii, Enterobacter oryzae, Klebsiella pneumoniae, Enterobacter dissolvens, milk-free solid Azospirillum amazonense and Cronobacter dublinensis were above 97%, respectively, indicating high diversity of endotoxin-fixing bacteria isolated.Physiological and biochemical determination of them showed that some strains With ammonia secretion capacity, production of indole, phosphorus, potassium solution, has certain potential in agricultural production.