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The differential strain virulence of the candidate toxins of Photorhabdus akhurstii can be correlated with their inter-strain gene sequence diversity

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Abstract

Photorhabdus akhurstii is an insect–parasitic bacterium that symbiotically associates with the nematode, Heterorhabditis indica. The bacterium possesses several pathogenicity islands that aids in conferring toxicity to different insects. Herein, we constructed the plasmid clones of coding sequences of four toxin genes (pirA, tcaA, tccA and tccC; each was isolated from four P. akhurstii strains IARI-SGMG3, IARI-SGGJ2, IARI-SGHR2 and IARI-SGMS1) in Escherichia coli and subsequently, their biological activity were investigated against the fourth-instar larvae of the model insect, Galleria mellonella via intra-hemocoel injection. Bioinformatics analyses indicated inter-strain amino acid sequence difference at several positions of the candidate toxins. In corroboration, differential insecticidal activity of the identical toxin protein (PirA, TcaA, TccA and TccC conferred 15–59, 27–100, 25–100 and 33–98% insect mortality, respectively, across the strains) derived from the different bacterial strains was observed, suggesting that the diverse gene pool in Indian strains of P. akhurstii leads to strain-specific virulence in this bacterium. These toxin candidates appear to be an attractive option to deploy them in biopesticide development for managing the insect pests globally.

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Funding

This study was supported by Science and Engineering Research Board (Grant Number YSS/2014/000452).

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Authors and Affiliations

  1. Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India

    Tushar K. Dutta, Chetna Mathur & Vishal S. Somvanshi

  2. Division of Agricultural Chemicals, ICAR-Indian Agricultural Research Institute, New Delhi, 110012, India

    Abhishek Mandal

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  1. Tushar K. Dutta
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Correspondence to Tushar K. Dutta.

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13205_2020_2288_MOESM1_ESM.pdf

Supplementary file1 Table S1. List of oligonucleotides employed for cloning and sequencing of Photorhabdus toxin genes. Annealing temperature for each PCR reaction was 60°C.Figure S1. Agarose gel photographs showing the PCR amplified fragments of toxin genes, such as pirA (417 bp), tcaA (3288 bp), tccA (2898 bp) and tccC (3132 bp) from the genomic DNA of P. akhurstii strains IARI-SGMG3, IARI-SGGJ2, IARI-SGHR2 and IARI-SGMS1.Figure S2. SDS-PAGE analysis of P. akhurstii (IARI-SGMG3) toxins induced by 1 mM IPTG for 4 h in E. coli BL21 cells. Lanes: M—protein molecular weight marker, 1—uninduced BL21 cells harboring TcaA, 2—induced BL21 cells harboring TcaA, 3—uninduced BL21 cells harboring TccC, 4—induced BL21 cells harboring TccC, 5—uninduced BL21 cells harboring TccA, 6—induced BL21 cells harboring TccA, 7—uninduced BL21 cells harboring PirA, 8—induced BL21 cells harboring PirA. Arrows indicate the overexpression of target protein. The calculated molecular mass (using Expasy ProtParam tool; https://web.expasy.org/protparam/) of TcaA, TccC, TccA and PirA was 120, 111, 105 and 15 kDa, respectively. (PDF 366 kb)

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Dutta, T.K., Mathur, C., Mandal, A. et al. The differential strain virulence of the candidate toxins of Photorhabdus akhurstii can be correlated with their inter-strain gene sequence diversity. 3 Biotech 10, 299 (2020). https://doi.org/10.1007/s13205-020-02288-0

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