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Characterisation of antagonistic Bacillus paralicheniformis (strain EAL) by LC–MS, antimicrobial peptide genes, and ISR determinants

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Abstract

Plants have their own defense mechanisms such as induced systemic resistance (ISR) and systemic-acquired resistance. Bacillus spp. are familiar biocontrol agents that trigger ISR against various phytopathogens by eliciting various metabolites and producing defense enzyme in the host plant. In this study, B. paralicheniformis (strain EAL) was isolated from the medicinal plant Enicostema axillare. Butanol extract of B. paralicheniformis showed potential antagonism against Fusarium oxysporum compared to control well (sterile distilled water) A liquid chromatography mass spectrometry analysis showed 80 different compounds. Among the 80 compounds, we selected citrulline, carnitine, and indole-3-ethanol based on mass-to-charge ratio, database difference, and resolution of mass spectrum. The synthetic form of the above compounds showed biocontrol activity against F. oxysporum under in vitro condition in combination, not as individual compounds. However, the PCR amplification of 11 antimicrobial peptide genes showed that none of the genes amplified in the strain. B. paralicheniformis inoculation challenged with F. oxysporum on tomato plants enhanced production of defense enzymes such as peroxidase (POD), superoxide dismutase (SOD), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and proline compared to control plants (without inoculation of B. paralicheniformis) at significant level (p < 0.005). Stem of tomato plants expressed higher POD (2.2-fold), SOD (2.2-fold), PPO (1.9-fold), and PAL (1.3-fold) contents followed by the leaf and root. Elevated proline accumulation was observed in the leaf (1.8-fold) of tomato plants. Thus, results clearly showed potentiality of B. paralicheniformis (EAL) in activation of antioxidant defense enzyme against F. oxysporum-infected tomato plants and prevention of oxidative damage though hydroxyl radicals scavenging activities that suppress the occurrence of wilt diseases.

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Acknowledgements

The authors thank UTU management for providing B. U. Patel research fellowship to carry out this work. Authors also acknowledge IIT Bombay SAIF for providing LC–MS analysis service.

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Authors and Affiliations

  1. C. G. Bhakta Institute of Biotechnology, Uka Tarsadia University, Maliba Campus, Bardoli, Surat, Gujarat, 394 350, India

    Hardik Naik Jinal & Natarajan Amaresan

  2. Division of Field Crop Improvement and Protection, Central Island Agricultural Research Institute, Port Blair, Andaman and Nicobar Islands, 744 101, India

    Krishnan Sakthivel

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  1. Hardik Naik Jinal
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  2. Krishnan Sakthivel
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  3. Natarajan Amaresan
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Contributions

NA: Designed the work. HNJ: Performed the experiments. KS: Performed AMP gene amplification work. HNJ and NA: Data analysis and drafting the article.

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Correspondence to Natarajan Amaresan.

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Jinal, H.N., Sakthivel, K. & Amaresan, N. Characterisation of antagonistic Bacillus paralicheniformis (strain EAL) by LC–MS, antimicrobial peptide genes, and ISR determinants. Antonie van Leeuwenhoek 113, 1167–1177 (2020). https://doi.org/10.1007/s10482-020-01423-4

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