Scientia Agricultura Sinica ›› 2007, Vol. 40 ›› Issue (1): 92-98 .doi: 10.3864/j.issn.0578-1752.at-2006-7557

• PLANT PROTECTION • Previous Articles     Next Articles

Application of ARDRA Technology to Isolation and Identification of Pseudomonas fluorescens

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  1. 中国农业科学院植物保护研究所
  • Received:2006-05-10 Revised:1900-01-01 Online:2007-01-10 Published:2007-01-10

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Abstract: A total of one hundred and sixty-three isolates of fluorescent Pseudomonas spp were isolated from 488 plant rhizospheric soil samples collected from Shandong, Hainan, Yunnan and Xinjiang Province. One hundred and two were detected to be rod and had monopolar one or more flagellum observed under light microscope. About 1.5 kb 16S rDNA fragment of these strains were amplified with the conservative 16S rDNA primer pairs of the eubacterium. The PCR products were purified and digested with the restriction enzyme HeaIII. Amplified ribosomal DNA restriction analysis (ARDRA) was performed. All strains produce three digestion patterns, which were similar with the digestion patterns of Pseudomonas fluorescens, Pseudomonas putida and unknown type. ARDRA analysis showed that all isolated strain belonged to at least three species. All the One hundred and two isolated strains were inhibitory ability to plant pathogenic fungi. Twenty-five of them showed inhibitory to Rhizoctonia solani in dual-culture test on PDA plates, one of them shown the highest antagonistic activity to Rhizoctonia solani was named TC222. Moreover, TC222 displayed inhibitory to various plant pathogens, e.g. Gaeumannomyces graminis, Magnaporthe grisea, Drechslera sorakiniana, Alternaria brassicae, Botrytis cinerea, Phomopsis asparagi, Fulvia fulva, Colletotrichum lindemuthianum Sacc. et Magn and Phytophthora parasitica var nicotianae. 16S rDNA sequence analysis of TC222 showed 99% homology to Pseudomonas fluorescens. TC222 was further confirmed to belong to Pseudomonas fluorescens according to physiological and biochemical characteristics.

Key words: Pseudomonas fluorescens, Isolation, ARDRA analysis, Antagonism

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