Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (7): 1303-1317.doi: 10.3864/j.issn.0578-1752.2012.07.008

• SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT • Previous Articles     Next Articles

Isolation and Phylogeny of Nitrogen-Fixing Endophytic Bacteria in Wheat, Rice, Maize, Chinese Cabbage and Celery

 SUN  Jian-Guang, LUO  Qiong, GAO  Miao, HU  Hai-Yan, XU  Jing, ZHOU  Yi-Qing   

  1. 中国农业科学院农业资源与农业区划研究所/农业部作物营养与施肥重点实验室，北京 100081
  • Received:2011-04-12 Online:2012-04-01 Published:2011-08-26

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Abstract: 【Objective】 The objective of this study is to determine the main groups and phylogenetic position of nitrogen-fixing endophytic bacteria in wheat, rice, maize, Chinese cabbage and celery. 【Method】 Surface sterilization and nitrogen-free medium were used to isolate nitrogen-fixing endophytic bacteria. Acetylene reduction assay was used to measure the nitrogenase activity. 16S rDNA was amplified by PCR and 16S rDNA sequence was analysed with MEGA software. 【Result】 Thirty four nitrogen-fixing endophytic bacteria with nitrogenase activity ranging from 0.30 to 30.24 nmol C2H4/h•mg protein were isolated from field wheat. Based on 16S rDNA sequence similarity, these strains belonged to 21 species of 13 genera, including Pseudomonas, Rhizobium, Bacillus, Flavobacterium, etc. Twenty five nitrogen-fixing endophytic bacteria with nitrogenase activity ranging from 3.12 to 254.12 nmol C2H4/h•mg protein were isolated from field rice. They belonged to 16 species of 9 genera, including Bacillus, Burkholderia, Enterobacter, Klebsiella, etc. Moreover, Burkholderia, Enterobacter and Klebsiella were dominent. Nine maize endophytic diazotrophs with nitrogenase activity ranging from 7.27 to 59.58 nmol C2H4/h•mg protein were identified as 6 species of genera Rhizobium, Sphingomonas, Arthrobacter, Brevibacterium and Microbacterium. Fourteen endophytic diazotrophs with nitrogenase activity ranging from 2.33 to 205.21 nmol C2H4/h•mg protein were isolated from pot trial pakchoi and identified as 8 species of 6 genera of Rhizobium, Arthrobacter, Pseudomonas, etc. Ten endophytic diazotrophs with nitrogenase activity ranging from 1.23 to 46.70 nmol C2H4/h•mg protein were isolated from marcket celery and identified as 8 species of 5 genera of Sphingomonas, Pseudomonas, Brevundimonas, etc. 【Conclusion】 Nitrogen-fixing endophytic bacteria colonized widely in wheat, rice, maize and vegetables. Their nitrogenase activity under pure culture ranged from 0.30 to 254.12 nmol C2H4/h•mg protein. Ninety two endophytic diazotrophs belonged to 56 species of 25 genera, including Pseudomonas, Rhizobium, Bacillus, Burkholderia, Sphingomonas, Enterobacter and Klebsiella, etc. These endophytic diazotrophs have a great potential to agriculture.

Key words: wheat, rice, maize, Chinese cabbage, celery, endophytic diazotrophs, phylogeny

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