Screening of Primer Sets for Amplification Cyclic Lipopeptide Gene of Bacillus Against Banana Fusarium Wilt
1 School of Life and Pharmaceutical Sciences, Hainan University, Haikou, 570228, China
2 Key Laboratory of Pests Comprehensive Governance for Tropical Crops, Ministry of Agriculture and Rural Affairs, Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, 571101, China
Molecular Microbiology Research, 2021, Vol. 11, No. 2 doi: 10.5376/mmr.2021.11.0002
Received: 16 Nov., 2021 Accepted: 17 Nov., 2021 Published: 28 Nov., 2021
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This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Deng T., Guo L.J., Yang L.Y., Zhou Y., Wang J., Liu Z.Q., and Huang J.S., 2021, Screening of primer sets for amplification cyclic lipopeptide gene of Bacillus against banana fusarium wilt, Molecular Microbiology Research, 11(2): 1-10 (doi: 10.5376/mmr.2021.11.0002)
Cyclic lipopeptides (CLPs) are a class of natural agricultural antibiotics produced by Bacillus, it plays an important role in the biological control using of Bacillus. In this study, 20 strains of Bacillus, including Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus siamese, and Bacillus cereus were determined the antagonistic ability against to pathogenic fungi such as Foc 4. PCR amplification was performed on 20 strains of Bacillus and 7 soil samples, by using the designed 14 pairs of primers for CLPs-related synthetase genes, including Bacillomycin, Fengycins, Iturins, Surfactin etc, screen CLPs primers that can be used for Bacillus and soil monitoring. The results showed that: a total of 16 Bacillus strains have the antagonistic activity against pathogens, and microscopic observation of the crude lipopeptide extract of fermentation broth of these Bacillus strains can destroy the biofilm system and cell wall of Foc4, causing the mycelium and meristems to expand and deform; 12 pairs primers of this study (BamD-F/R, BamA-F/R, FenA-F/R, FenB-F/R, 61FenA-F/R, 61FenB-F/R, ItuA-F/R, ItuB-F/R, ItuC-F/R, ItuD-F/R, SrfAA-F/R, SrfAB-F/R) could effective amplify CLPs gene, and find that all of test Bacillus strains has the ItuC gene, there is a positive relationship between the antagonistic ability of Bacillus and the diversity and richness of CLPs genes, through PCR amplification of total soil DNA, it was found that the designed CLPs primers were feasible to monitor Bacillus in soil environment. It would be used as an important supplement to conventional plate counting and high-throughput sequencing methods, monitor and guide the application of Bacillus control soil-borne diseases of plants to provide theoretical support for further field.
Banana Fusarium Wilt; Antagonistic Bacillus; Cyclic lipopeptide antibiotic gene