Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (20): 4064-4076.doi: 10.3864/j.issn.0578-1752.2015.20.008

• PLANT PROTECTION • Previous Articles     Next Articles

The Antifungal Activities of Bacillus spp. and Its Relationship with Lipopeptide Antibiotics Produced by Bacillus spp.

 XIANG Ya-ping1,2, CHEN Zhi-yi1,2, LUO Chu-ping1,2, ZHOU Hua-fei1,2, LIU Yong-feng2
  

  1. 1College of Plant Protection, Nanjing Agricultural University, Nanjing 210095
    2Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014
  • Received:2015-04-13 Online:2015-10-20 Published:2015-10-20

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Abstract:

【Objective】 The objective of this study is to clear the inhibition effect of 55 Bacillus spp. strains to pathogenic fungi, Rhizoctonia cerealis, Fusarium graminearum and Fusarium oxysporum f. sp. niveum, and the qualitative and quantitative relationship withlipopeptide antibiotics produced by the Bacillus strains.【Method】Inhibition rates of the 55 Bacillus spp. strains and their crude lipopeptides extracts against three pathogenic fungi were tested, respectively, by dual culture on plate. Three pairs of primers designed based on the known lipopeptide genes were used to amplify the corresponding genes, srfA, ituA and fenA, from the genome of the 55 strains. The lipopeptides antibiotics were identified by high performance liquid chromatography (HPLC) and mass spectrum (MS) techniques.【Result】 The 55 strains of Bacillus spp. and their crude lipopeptides extracts had certain biological activities to the tested pathogenic fungi. The crude lipopeptides extracts of 16.5% strains had higher antifungal activities than the Bacillus strains in vivo, the antifungal activities against the pathogenic fungi of crude lipopeptides extracts of 55.8% strains were weaker than living cells of the strains in vivo, while the antifungal activities of 27.7% strains did not change basically. The results of PCR amplification showed that among the 55 Bacillu strains, the genomes of 89.1% Bacillus strains contained srfA, 87.3% strains contained ituA, only 56.4% strains contained fenA. Among the 55 Bacillus strains, 53 Bacillus strains were identified with surfactins in the crude lipopeptides extracts by the MS analysis, 48 strains were identified with iturins, only Bs916 (B. subtilis 916) was identified having fenfycins in the crude lipopeptides extracts. Those strains that were not identified having the three kinds of lipopeptides had weaker or no antifungal activity. There were 48 strains which were found to have iturins all identified with bacillomycin D, only 2 strains were identified with bacillomycin L, 12 strains were identified with bacillomycin F, iturin A or mycosubtilin. Among the 55 Bacillus strains, 53 strains were detected with suefactins by HPLC, 51 strains were detected with iturins, only Bs916 strain was detected with fengycins and 2 strains were not detected with the 3 kinds of lipopeptide antibiotics. According to the time and type of chromatographic peak, iturins were divided into 4 types, named as types 1, 2, 3, and 4. The antifungal activities intensity of 4 types of iturins were type 1＞type 2＞type 3＞type 4. There were two unknown lipopeptides (named as Unknowns: Unknown1, 2 ) which were speculated as new lipopeptides were detected with MS analysis in 3 strains JCC-1, and JCC-9 and JCC-18. 【Conclusion】 Lipopeptides antibiotics produced by Bacillus spp. were the main composition against plant pathogenic fungi and had diversity characteristics. The structure changes and secretion of iturins had certain relevance to antifungal activities. There were some unknown new resisted resources in lipopeptides compounds from Bacillus spp..

Key words: Bacillus spp., antifungal activity, lipopeptides, correlation

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