Journal of Biological Control

Open Access Open Access  Restricted Access Subscription Access

Biocontrol Potential and Molecular Characterization of Lipopeptides Producing Bacillus Subtilis Against Sclerotinia Sclerotiorum


Affiliations
1 Department of Agricultural Microbiology, UAS, GKVK, Bengaluru – 560 065, Karnataka, India
2 ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
3 Department of Plant Pathology, UAS, GKVK, Bengaluru – 560065, Karnataka, India
4 Rice Pathology Laboratory ARS, Gangavathi, UAS Raichur – 584104, Karnataka, India
 

Bacillus subtilis  is  a Gram-positive and endospore producing bacterium. Limited studies have shown that lipopeptides produced by  B. subtilis  can be inhibitory to phytopathogens.   Sclerotinia sclerotiorum  is a plant pathogenic fungus  which causes  various diseases like cotton rot, watery soft rot, stem rot, crown rot and blossom blight in vegetable crops. The objective of the study was to isolate lipopeptides from  B. subtilis  and study their inhibitory potential against  S. sclerotiorum. So, the B. subtilis  isolates  were extracted from the collected soils of Western Ghats of India. They were initially characterized through morphological parameters followed by PCR amplification of the 16S rDNA gene and confirmation through BLAST algorithm in NCBI database. The lipopeptides produced by these isolates were tested against  S. sclerotiorum. B. subtilis  strains were effective against  S. sclerotiorum and exhibited 18.33 to 29.5 % inhibition under dual culture bio-assay. The antagonistic activity of lipopeptides extracted from  B. subtilis  strains showed 21.56 to 88.89 % inhibition of  S. sclerotiorum  in the lowest to highest concentration of lipopeptide tested and was found to be significantly higher than the control. The present study has shown that  B. subtilis  strains vary in the production of lipopeptides and some of them could produce lipopeptides that are highly inhibitory to S. sclerotiorum. B. subtilis  strain NBAIR BSWG1 showed the highest inhibition for S. sclerotiorum. Lipopeptide based poison food technique and the dual culture bioassay results showed that B. subtilis  strain NBAIR BSWG1 has immense potential for use in the biological control of  S. sclerotiorum. Further studies are being carried out in formulating the lipopeptides for field application.

Keywords

Antimicrobial property, biopesticide, PCR, soft rot of vegetable, Western Ghats.
User
Notifications

  • Abriouel, H., Franz, C. M., Omar, N. B. and Galvez, A. 2011. Diversity and applications of Bacillus bacteriocins. FEMS Microbiol Rev, 35: 201-232. https://doi.org/10.1111/j.1574-6976.2010.00244.x

  • Biniarz, P., Lukaszewicz, M. and Janek, T. 2017. Screening concepts, characterization and structural analysis of microbial-derived bioactive lipopeptides a review. Crit Rev Biotechnol, 37(3): 393-410. https://doi.org/10.3109/07388551.2016.1163324

  • Gao, H., Xu, X., Dai, Y. and He, H. 2016. Isolation, identification and characterization of Bacillus subtilis CF-3, a bacterium from fermented bean curd for controlling postharvest diseases of peach fruit. Food Sci Technol Res, 22(3): 377-385. https://doi.org/10.3136/fstr.22.377

  • Gautham, S. A., Shobha, K. S., Onkarappa, R. and Kekuda, T. R. 2012. Isolation, character isation and antimicrobial potential of Streptomyces species from Western Ghats of Karnataka, India Res J Pharm Technol, 5(2): 233-238.

  • Hashem, A., Tabassum, B. and Allah, E. F. A. 2019. Bacillus subtilis: a plant growth promoting rhizobacterium that also impacts biotic stress. Saudi J Biol Sci, 26: 1291-1297. https://doi.org/10.1016/j.sjbs.2019.05.004

  • RUQIYA et al.

  • Jeyaseelan, E. C., Tharmila, S. and Niranjan, K. 2012. Antagonistic activity of Trichoderma spp. and Bacillus spp. against Pythium aphanidermatum isolated from tomato damping off. Arch Appl Sci Res, 4(4): 1623-1627.

  • Jiang, J., Gao, L., Bie, X., Lu, Z., Liu, H., Zhang, C., Lu, F. and Zhao, H. 2016. Identification of novel surfactin derivatives from NRPS modification of Bacillus subtilis and its antifungal activity against Fusarium moniliforme. BMC Microbiol, 16: 31. https://doi.org/10.1186/s12866-016-0645-3

  • Johnson, J. S., Spakowicz, D. J., Hong, B. Y., Petersen, L. M., Demkowicz, P., Chen, L., Leopold, S. R., Hanson, B. M., Agresta, H. O., Gerstein, M. and Sodergren, E. 2019. Evaluation of 16S rRNA gene sequencing for species and strain-level microbiome analysis. Nature Communications, 10(1): 5029. https://doi.org/10.1038/s41467-019-13036-1

  • Kaur, P. K., Joshi, N., Singh, I. P. and Saini, H. S. 2016. Identification of cyclic lipopeptides produced by Bacillus vallismortis R2 and their antifungal activity against Alternaria alternate. J Appl Microbiol, 122: 139-152.

  • Kumbar, B., Mahmood, R. and Narasimhappa, N. S. 2017. Identification and molecular diversity analysis of Bacillus subtilis from soils of Western Ghats of Karnataka using 16S rRNA bacterial universal primers. Int J Pure App Biosci, 5(2): 541-548. https://doi.org/10.18782/2320-7051.2721

  • Leelasuphakul, W., Hemmanee, P. and Chuenchitt, S. 2008. Growth Inhibitory properties of Bacillus subtilis strains and their metabolites against the green mold pathogen (Penicillium digitatum Sacc.) of citrus fruit. Postharvest Biol Technol, 48(1): 113-121. https://doi.org/10.1016/j.postharvbio.2007.09.024

  • Li, X. Y., Mao, Z. C., Wang, Y. H., Wu, Y. X., He, Y. Q. and Long, C. L. 2012. ESI LCMS and MS/MS characterization of antifungal cyclic lipopeptides produced by Bacillus subtilis XF-1. Adv Microbial Physiol, 22(2): 83-93. https://doi.org/10.1159/000338530

  • Ma, Y., Kong, Q., Qin, C., Chen, Y., Chen, Y., Lv, R. and Zhou, G. 2016. Identification of lipopeptides in Bacillus megaterium by two-step ultrafiltration and LC–ESI–MS/MS. Amb Express 6(1): 1-15. https://doi.org/10.1186/s13568-016-0252-6

  • Mardanova, A. M., Hadieva, G. F., Lutfullin, M. T., Khilyas, I. V., Minnullina, L. F., Gilyazeva, A. G., Bogomolnaya, L. M. and Sharipova, M. R. 2016. Bacillus subtilis strains with antifungal activity against the phytopathogenic fungi. Agric Sci, 8(1): 1-20. https://doi.org/10.4236/as.2017.81001

  • Miljkovic, M., Jovanovic, S., O’Connor, P. M., Mirkovic, N., Jovcic, B. and Filipic, B. 2019. Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials. PLoS One 14(5): e0216773. https://doi.org/10.1371/journal.pone.0216773

  • Penha, R. O., Vandenberghe, L. P., Faulds, C., Soccol, V. T. and Soccol, C. R. 2020. Bacillus lipopeptides as powerful pest control agents for a more sustainable and healthy agriculture: Recent studies and innovations. Planta, 251: 1-5. https://doi.org/10.1007/s00425-020-03357-7

  • Perez, R. H., Zendo, T. and Sonomoto, K. 2018. Circular and leaderless bacteriocins: Biosynthesis, mode of action, applications and prospects. Front Microbiol, 9: 2085. https://doi.org/10.3389/fmicb.2018.02085

  • Schloss, P. D. and Handelsman, J. 2005. Introducing DOTUR, a computer program for defining operational taxonomic units and estimating species richness. Appl Environ Microbiol 71: 1501. https://doi.org/10.1128/AEM.71.3.1501-1506.2005

  • Sharma, D., Singh, S. S., Baindara, P., Sharma, S., Khatri, N., Grover, V., Patil, P. B. and Korpole, S. 2020. Surfactin like broad Spectrum antimicrobial lipopeptide co-produced with sublancin from Bacillus subtilis Strain A52: Dual reservoir of bioactives. Front Microbiol, 11: 1167. https://doi.org/10.3389/fmicb.2020.01167

  • Sicuia, O. A., Olteanu, V., Ciuca, M., Cîrstea, D. M. and Cornea, C. P. 2011. Characterization of new Bacillus spp. isolates for antifungal properties and biosynthesis of lipopeptides. Sci Papers Ser A Agron, 54: 482-491.

  • Yu, X., Ai, C., Xin, L. and Zhou, G. 2011. The siderophore-producing bacterium, Bacillus subtilis CAS15, has a biocontrol effect on Fusarium wilt and promotes the growth of pepper. Eur J Soil Sci, 47(2): 138-145. https://doi.org/10.1016/j.ejsobi.2010.11.001

  • Zaccardelli, M., Sorrentino, R., Caputo, M., Scotti, R., De Falco, E. and Pane, C. 2020. Stepwise-selected Bacillus amyloliquefaciens and Bacillus subtilis strains from composted aromatic plant waste able to control soil-borne diseases. Agri, 10(2): 30. https://doi.org/10.3390/agriculture10020030

  • Zhang, L. and Sun, C. 2018. Fengycins, cyclic lipopeptides from marine Bacillus subtilis strains, kill the plant-pathogenic fungus Magnaporthe grisea by inducing reactive oxygen species production and chromatin condensation. Appl Environ Microbiol, 84(18). https://doi.org/10.1128/AEM.00445-18


Abstract Views: 149

PDF Views: 96




  • Biocontrol Potential and Molecular Characterization of Lipopeptides Producing Bacillus Subtilis Against Sclerotinia Sclerotiorum

Abstract Views: 149  |  PDF Views: 96

Authors

S. RUQIYA
Department of Agricultural Microbiology, UAS, GKVK, Bengaluru – 560 065, Karnataka, India
H. C. GIRISHA
Department of Agricultural Microbiology, UAS, GKVK, Bengaluru – 560 065, Karnataka, India
C. MANJUNATHA
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
R. RANGESHWARAN
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
A. KANDAN
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
G. SIVAKUMAR
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
M. K. PRASANNA KUMAR
Department of Plant Pathology, UAS, GKVK, Bengaluru – 560065, Karnataka, India
D. PRAMESH
Rice Pathology Laboratory ARS, Gangavathi, UAS Raichur – 584104, Karnataka, India
K. T. SHIVAKUMARA
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
H. S. VENU
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
S. NANDITHA
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
K. S. ANKITHA
Department of Agricultural Microbiology, UAS, GKVK, Bengaluru – 560 065, Karnataka, India
K. ADITYA
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
N. AARTHI
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India
S. N. SUSHIL
ICAR-National Bureau of Agricultural Insect Resources, Bengaluru – 560024, Karnataka, India

Abstract


Bacillus subtilis  is  a Gram-positive and endospore producing bacterium. Limited studies have shown that lipopeptides produced by  B. subtilis  can be inhibitory to phytopathogens.   Sclerotinia sclerotiorum  is a plant pathogenic fungus  which causes  various diseases like cotton rot, watery soft rot, stem rot, crown rot and blossom blight in vegetable crops. The objective of the study was to isolate lipopeptides from  B. subtilis  and study their inhibitory potential against  S. sclerotiorum. So, the B. subtilis  isolates  were extracted from the collected soils of Western Ghats of India. They were initially characterized through morphological parameters followed by PCR amplification of the 16S rDNA gene and confirmation through BLAST algorithm in NCBI database. The lipopeptides produced by these isolates were tested against  S. sclerotiorum. B. subtilis  strains were effective against  S. sclerotiorum and exhibited 18.33 to 29.5 % inhibition under dual culture bio-assay. The antagonistic activity of lipopeptides extracted from  B. subtilis  strains showed 21.56 to 88.89 % inhibition of  S. sclerotiorum  in the lowest to highest concentration of lipopeptide tested and was found to be significantly higher than the control. The present study has shown that  B. subtilis  strains vary in the production of lipopeptides and some of them could produce lipopeptides that are highly inhibitory to S. sclerotiorum. B. subtilis  strain NBAIR BSWG1 showed the highest inhibition for S. sclerotiorum. Lipopeptide based poison food technique and the dual culture bioassay results showed that B. subtilis  strain NBAIR BSWG1 has immense potential for use in the biological control of  S. sclerotiorum. Further studies are being carried out in formulating the lipopeptides for field application.

Keywords


Antimicrobial property, biopesticide, PCR, soft rot of vegetable, Western Ghats.

References





DOI: https://doi.org/10.18311/%2Fjbc%2F2023%2F33785