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Expression of Heterorhabditis bacteriophora C-type lectins, Hb-clec-1 and Hb-clec-78, in context of symbiosis with Photorhabdus bacteria

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Abstract

Insect-parasitic nematodes of the genus Heterorhabditis live in a symbiotic relationship with a gram negative Gamma-proteobacteria of the genus Photorhabdus. This nematode-bacteria pair is a simple and genetically tractable model to study animal-microbe symbiosis. Here we investigated the role of Heterorhabditis nematode C-type lectin (clec) genes in context of nematode-bacteria symbiosis. The in silico analysis identified seven clec genes in H. bacteriophora and three clec genes in H. indica. Two of the clec genes, H. bacteriophora clec-1 (Hb-clec-1) and H. bacteriophora clec-78 (Hb-clec-78) were further characterized. Both of these genes were present in a single copy in the H. bacteriophora genome. The phylogenetic analysis revealed that H. bacteriophora CLEC proteins were close to CLEC-1 and CLEC-78 proteins of free living Caenorhabditis but not to the CLEC proteins of insect-parasitic Steinernema nematodes which share a similar symbiotic relationship with Xenorhabdus bacteria. In situ hybridization showed that expression of Hb-clec-1 and Hb-clec-78 was localized to the alimentary canal of infective juveniles (IJs) in the region of terminal bulb, oesophago-intestinal valve and anterior part of intestine. Hb-clec-78 gene expression displayed significant positive correlation to the presence of bacteria during various stages of symbiosis: it was up-regulated during all the nematode developmental stages when Photorhabdus was symbiotically associated, but down-regulated at the post-IJ recovery stage when the developing nematodes were free of bacteria. Hb-clec-1 gene expression did not show any correlation with presence or absence of symbiont bacteria. Subject to genetic validation, our study suggests that Hb-clec-78 might be actively involved in modulation of symbiosis with Photorhabdus symbionts.

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Fig. 1: Protein domain and copy number analysis of Heterorhabditis clec sequences.
Fig. 2: Molecular Phylogenetic analysis of HB-CLEC-1 and HB-CLEC-78 proteins by Maximum Likelihood (ML) method.
Fig. 3: The symbiosis-relevant developmental stages ofH. bacteriophoranematodes and the expression ofHb-clec-1andHb-clec-78.
Fig. 4: In situ hybridization of gene specific DIG-labelled probes to determine the site of expression of Hb-clec-1 and Hb-clec-78 genes in the infective juveniles of Heterorhabditis bacteriophora .

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Acknowledgements

M.Sc. student CGB acknowledges the Junior Research Fellowship from the Indian Council of Agricultural Research, and PG School, ICAR-Indian Agricultural Research Institute, New Delhi. This work was supported by funding from Science and Engineering Research Board, Department of Science and Technology, Government of India [Grant no. SB/SO/AS/010/2014 to VSS], and in-house funding from the Division of Nematology, ICAR-Indian Agricultural Research Institute, New Delhi.

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Correspondence to Uma Rao or Vishal Singh Somvanshi.

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Bhat, C.G., Chauhan, K., Phani, V. et al. Expression of Heterorhabditis bacteriophora C-type lectins, Hb-clec-1 and Hb-clec-78, in context of symbiosis with Photorhabdus bacteria. Symbiosis 77, 49–58 (2019). https://doi.org/10.1007/s13199-018-0569-9

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  • DOI: https://doi.org/10.1007/s13199-018-0569-9

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