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Frontiers of Agricultural Science and Engineering

ISSN 2095-7505

ISSN 2095-977X(Online)

CN 10-1204/S

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Front. Agr. Sci. Eng.    2018, Vol. 5 Issue (2) : 262-270    https://doi.org/10.15302/J-FASE-2017166
RESEARCH ARTICLE
Fusaricidins in Paenibacillus polymyxa A21 and their antagonistic activity against Botrytis cinerea on tomato
Weicheng LIU1, Xiaoli WU2, Xuelian BAI1, Hong ZHANG1, Dan DONG1, Taotao ZHANG1, Huiling WU1()
1. Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
2. Editorial Department of Chinese Science Abstracts, Science and Technology Review Publishing House, Beijing 100081, China
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Abstract

Four kinds of antifungal compounds from an extract of Paenibacillus polymyxa A21 with molecular masses of 883.56, 897.59, 947.55, and 961.58 Da were characterized as the members of fusaricidin-type of antibiotics according to LC-MS analysis. Fusaricidins isolated from culture filtrate displayed high antagonistic activity against several plant fungal pathogens, especially Botrytis cinerea, the causal agent of gray mold. The fusaricidins biosynthetic gene cluster (BGC) from A21 was cloned by PCR and comparative cluster analysis revealed that gene fusTE, the 3′ boundary of the fusaricidin BGC in strain PKB1, was not present in fusaricidin BGC of A21, indicating that fusTE is not necessary for fusaricidin synthesis. Fusaricidin extract from A21 significantly reduced gray mold disease incidence and severity on tomato. The mRNA levels for three patho-genesis-related proteins (PRs) revealed that treatment of tomato leaves with fusaricidin extract induced the expression of PR genes to different levels, suggesting that one reason for the reduction of gray mold infection by fusaricidin is induction of PR proteins, which lead to increased resistance to pathogens. This is the first report of the application of fusaricidins to control tomato gray mold and the comparative cluster analysis provides important molecular basis for research on fusaricidin biosynthesis.
Keywords antifungal activity      biosynthetic gene cluster      Botrytis cinerea      fusaricidin      Paenibacillus polymyxa     
Corresponding Author(s): Huiling WU   
Just Accepted Date: 27 September 2017   Online First Date: 31 October 2017    Issue Date: 28 May 2018
 Cite this article:   
Weicheng LIU,Xiaoli WU,Xuelian BAI, et al. Fusaricidins in Paenibacillus polymyxa A21 and their antagonistic activity against Botrytis cinerea on tomato[J]. Front. Agr. Sci. Eng. , 2018, 5(2): 262-270.
 URL:  
https://academic.hep.com.cn/fase/EN/10.15302/J-FASE-2017166
https://academic.hep.com.cn/fase/EN/Y2018/V5/I2/262
Fungal pathogens Average diameter of inhibition zone/cm
Botrytis cinerea 3.43±0.08
Glomerella cingulata 3.16±0.14
Botrytis cinerea 2.66±0.15
Fusarium oxysporum f. sp. niveum 2.62±0.10
F. graminearum 2.34±0.07
Monilia cinerea 1.93±0.09
Vermicularia capsici 1.93±0.13
F. oxysporum f. sp. vasinfectum 1.65±0.06
Rhizoctonia cerealis 1.62±0.03
F.oxysporum f. sp. cucumerinum 1.62±0.12
F. solani 1.16±0.11
F. oxysporum f. sp. conglutinan 1.12±0.04
Tab.1  Antifungal activities of the crude extract of Paenibacillus polymyxa strain A21
Fig.1  RP-HPLC peak profile of antifungal compounds from Paenibacillus polymyxa A21. (a) Antifungal activity was found in the peak eluted at 24.5 min, as indicated by the arrow; (b) mass spectroscopic analysis of antifungal compounds produced by P. polymyxa A21. Compounds from peaks with molecular weights 947.56, 961.58, 883.56 and 897.59 Da (M+ H)+ showed antifungal activity. Mass spectroscopy was done in both positive and negative ion modes.
Fig.2  Analysis of fusaricidin biosynthetic gene clusters (BCGs). (a) BGCs of Paenibacillus polymyxa A21; (b) comparison of the fusaricidin BGCs of afur, sfur and pfur clusters using the MAUVE program. The horizontal panels depict the afur (upper), sfur (middle) and pfur (lower) clusters. Mean sequence similarities are proportional to the heights of the red bars, and the respective scales show the sequence coordinates in base pairs. Regions with low similarities and strain-specific regions are marked with black triangles. Arrows indicate deduced transcriptional units.
Treatment Disease incidence/% Average diameter of lesion/mm2
Fusaricidin extract from A21 35±2.4 b 7.7±0.4 b
Botrytis cinerea inoculated control 100±0.0 a 30.2±2.3 a
Boscalid 32±1.9 b 6.9±0.2 b
Blank control 0±0.0 c 0±0.0 c
Tab.2  Antagonistic activity of the fusaricidin extract from Paenibacillus polymyxa strain A21 against Botrytis cinerea infection of tomato leaves
Fig.3  Relative gene expression of defense-related genes PR-1, PR-2 and PR-3 in tomato leaves treated with fusaricidin analyzed by qRT-PCR (Fus, fusaricidin treatment before inoculation with B. cinerea; CK, inoculated control). Y-axes scale is relative expression level. Error bars indicate±SD.
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