Abstract
Bacillus sp. strain M10 was observed to produce an antifungal protein that inhibits the growth of Colletotrichum capsici, which is the causal agent of anthracnose disease of chili pepper and tomato. Ammonium sulfate precipitation, anion exchange chromatography, and sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the protein was approximately 55.4 kDa. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis and a subsequent sequence database search indicated the antifungal protein was most similar to the Bacillus amyloliquefaciens vegetative catalase (KatA) protein. Light microscopy observation revealed that the antifungal protein induced abnormal hyphal elongation and conidial swelling and rupture. The protein considerably inhibited anthracnose development and protected the fruits from C. capsici infection. Thus, Bacillus sp. strain M10 and/or its putative catalase may be useful as a post-harvest biocontrol agent that protects chili pepper and tomato fruits from anthracnose disease caused by C. capsici.
Similar content being viewed by others
References
Alina SO, Constantinscu F, Petruta CC (2015) Biodiversity of Bacillus subtilis group and beneficial traits of Bacillus species useful in plant protection. Rom Biotech Lett 20:10737–10750
Alvindia DG, Acda MA (2015) The antagonistic effect and mechanisms of Bacillus amyloliquefaciens DGA14 against anthracnose in mango cv. ‘Carabao’. Biocontrol Sci Technol 25:560–572
Avis TJ (2007) Antifungal compounds that target fungal membranes: applications in plant disease control. Can J Plant Pathol 29:323–329
Barefoot SF, Klaenhammer TR (1983) Detection and activity of lactacin B, a bacteriocin produced by Lactobacillus acidophilus. Appl Environ Microbiol 45:1808–1815
Byrne JM, Hausbeck MK, Hammerschmidt R (1997) Conidial germination and appressorium formation of Colletotrichum coccodes on tomato foliage. Plant Dis 81:715–718
Chowdhury SP, Hartmann A, Gao X, Borriss R (2015) Biocontrol mechanism by root-associated Bacillus amyloliquefaciens FZB42—a review. Front Microbiol 6:780. https://doi.org/10.3389/fmicb.2015.00780
Diao YZ, Zhang C, Xu JP, Lin D, Liu L, Mtung’e OG, Liu XL (2015) Genetic differentiation and recombination among geographic populations of the fungal pathogen Colletotrichum truncatum from chili peppers in China. Evol Appl 8:108–118
Dieguez-Uribeondo J, Forster H, Soto-Estrada A, Adaskaveg JE (2005) Subcuticular-intracellular hemibiotrophic and intercellular necrotrophic development of Colletotrichum acutatum on almond. Phytopathology 95:751–758
During K, Porsch P, Mahn A, Brinkmann O, Gieffers W (1999) The non-enzymatic microbicidal activity of lysozymes. FEBS Lett 449:93–100
El-Awady M, Moghaieb EEA, Haggag W, Sawsan Youssef S, El-Sharkawy AM (2008) Transgenic canola plants over-expressing bacterial catalase exhibit enhanced resistance to Peronospora parasitica and Erysiphe polygoni. Arab J Biotechnol 11:71–84
Hou X, Boyetchko SM, Brkic M, Olson D, Ross A, Hegedus D (2006) Characterization of the anti-fungal activity of a Bacillus spp. associated with sclerotia from Sclerotinia sclerotiorum. Appl Microbiol Biotechnol 72:644–653
Kim BS, Hwang BK (2007) Microbial fungicides in the control of plant diseases. J Phytopathol 155:641–653
Kim PI, Ryu J, Kim YH, Chi YT (2010) Production of biosurfactant lipopeptides iturin A, fengycin and surfactin A from Bacillus subtilis CMB32 for control of Colletotrichum gloeosporioides. J Microbiol Biotechnol 20:138–145
Kim JD, Jeon BJ, Han JW, Park MY, Kang SA, Kim BS (2016) Evaluation of the endophytic nature of Bacillus amyloliquefaciens strain GYL4 and its efficacy in the control of anthracnose. Pest Manag Sci 72:1529–1536
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lavermicocca P, Lonigro SL, Evidente A, Andolfi A (1999) Bacteriocin production by Pseudomonas syringae pv. ciccaronei NCPPB2355. Isolation and partial characterization of the antimicrobial compound. J Appl Microbiol 86:257–265
Liao CY, Chen MY, Chen YK, Kuo KC, Chung KR, Lee MH (2012) Formation of highly branched hyphae by Colletotrichum acutatum within the fruit cuticles of Capsicum spp. Plant Pathol 61:262–270
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Mataragas M, Metaxopoulos J, Galiotou M, Drosinos EH (2003) Influence of pH and temperature on growth and bacteriocin production by Leuconostoc mesenteroides L124 and Lactobacillus curvatus L442. Meat Sci 64:265–271
McLean KS, Roy KW (1991) Weeds as a source of Colletotrichum capsici causing anthracnose on tomato fruit and cotton seedlings. Can J Plant Pathol 13:131–134
Meerak J, Lida H, Watanabe Y, Miyashita M, Sato H, Nakagawa Y, Tahara Y (2007) Phylogeny of gamma-polyglutamic acid-producing Bacillus strains isolated from fermented soybean foods manufactured in Asian countries. J Gen Appl Microbiol 53:315–323
Nagorska K, Bikowski M, Obuchowski M (2007) Multicellular behaviour and production of a wide variety of toxic substances support usage of Bacillus subtilis as a powerful biocontrol agent. Acta Biochim Pol 54:495–508
Ni J, Tokuyama S, Sogabe A, Kawamura Y, Tahara Y (2001) Cloning and high expression of catalase gene from Bacillus sp. TE124. J Biosci Bioeng 91:422–424
Rais A, Jabeen Z, Shair F, Hafeez FY, Hassan MN (2017) Bacillus spp., a bio-control agent enhances the activity of antioxidant defense enzymes in rice against Pyricularia oryzae. PLoS ONE 12(11):e0187412
Selitrennikoff CP (2001) Antifungal proteins. Appl Environ Microbiol 67:2883–2894
Shafi J, Tian H, Ji MS (2017) Bacillus species as versatile weapons for plant pathogens: a review. Biotechnol Biotec Eq 31:446–459
Sharma RR, Singh D, Singh R (2009) Biological control of postharvest diseases of fruits and vegetables by microbial antagonists: a review. Biol Control 50:205–221
Shuqing L, Nan Z, Zhenhua Z, Jia L, Biao S, Ruifu Z, Qirong S (2013) Antagonist Bacillus subtilis HJ5 controls Verticillium wilt of cotton by root colonization and biofilm formation. Biol Fert Soils 49:295–303
Wang LT, Lee FL, Tai CJ, Kasai H (2007) Comparison of gyrB gene sequences, 16S rRNA gene sequences and DNA-DNA hybridization in the Bacillus subtilis group. Int J Syst Evol Microbiol 57:1846–1850
Wang N, Liu M, Guo L, Yang X, Qiu D (2016) A novel protein elicitor (PeBA1) from Bacillus amyloliquefaciens NC6 induces systemic resistance in tobacco. Int J Biol Sci 12:757–767
Xiao ZJ, Lu JR, Ma CQ, Xu P (2009) Formation and identification of trimethylimidazole during tetramethylpyrazine production from glucose by Bacillus strains. Biotechnol Lett 31:1421–1425
Yamamoto S, Shiraishi S, Suzuki S (2015) Are cyclic lipopeptides produced by Bacillus amyloliquefaciens S13-3 responsible for the plant defence response in strawberry against Colletotrichum gloeosporioides? Lett Appl Microbiol 60:379–386
Acknowledgement
This work was supported by The 90th Anniversary of Chulalongkorn University Fund (Ratchadaphiseksomphot Endowment Fund).
Author information
Authors and Affiliations
Corresponding author
Additional information
Handling Editor: Jesus Mercado Blanco.
Rights and permissions
About this article
Cite this article
Srikhong, P., Lertmongkonthum, K., Sowanpreecha, R. et al. Bacillus sp. strain M10 as a potential biocontrol agent protecting chili pepper and tomato fruits from anthracnose disease caused by Colletotrichum capsici. BioControl 63, 833–842 (2018). https://doi.org/10.1007/s10526-018-9902-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10526-018-9902-8