EVALUATION OF CHITINASE AS SYNERGIST IN THE MANAGEMENT OF TICK
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Date
2021
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Abstract
Chitinases play an important role in tick physiology and also have a proven role as bioacaricide. In the current study, an effort was made to amplify chitinase 1 genes of R. sanguineus and R. (B.) microplus from the pre-hatch stage of their eggs. The degenerative primers designed to amplify Haemaphysalis longicornis chitinase 1 gene failed to amplify. Amplification of putative chitinases of R. sanguineus and R. (B.) microplus ticks was attempted by designing a new set of primers for the reference sequences and amplicons of R. sanguineus and R. (B.) microplus chitinases were observed at 1320 and 1305 bp respectively.
The amplicons were subjected for sequencing and phylogenetic analysis which confirmed their identity (NCBI accession no. MT635600 and MT635601). MT635600 sequence revealed 28 nucleotide substitutions when compared to the reference sequence (GQ215231) with 97.87 per cent homology. MT635601 showed 97.93 per cent homology with reference shot gun assembly sequence of R. (B.) microplus putative chitinase (GBBR01000100). A total of 21 nucleotide substitutions and 6 nucleotide deletion from 143 to 148 was observed in the newly obtained sequence of putative chitinase of R. (B.) microplus when aligned with the reference sequence. The amino acid sequences were also compared pairwise and R. sanguineus putative chitinase showed 99.32 per cent identity to the reference sequence. The R. (B.) microplus putative chitinase amino acid sequence showed 97.93 per cent identity to the reference sequence. In R. sanguineus putative chitinase 19 O- and 3 N-glycosylation sites were observed in comparison to 17 O- and 3 N-glycosylation sites observed in R. (B.) microplus putative chitinase.