Mutational Analysis of Cadherin and v-ATPase Genes in Pink Bollworm and Resistance Towards Bt Cotton Through In-Vitro Insect Feeding Assay
Mutational Analysis of Cadherin and v-ATPase Genes in Pink Bollworm and Resistance Towards Bt Cotton Through In-Vitro Insect Feeding Assay
Ejaz Ali1*, Shahbaz Ahmed2, Bushra Tabassum1,2, Uzma Qaisar2 and Idrees Ahmad Nasir1,3
ABSTRACT
Insect pest attack is one of the major problems in the agriculture sector. Bt crops were found effective in providing resistance against pink bollworm in cotton but recent reports on insect-evolved resistance have highlighted mutations in pink bollworm (PBW) receptor genes. We investigated cadherin and v-ATPase genes from local isolates of PBW for possible mutations. We amplified ~1224bp and ~1149bp cadherin and v-ATPase genes respectively and cloned them in the pJET2.1 vector. The deduced sequence of cadherin didn’t exhibit any nucleotide change while v-ATPase gene showed mutations and was submitted in GenBank under accession No. OR231877. The CDS of the v-ATPase showed four mutations; three silent and one missense mutation. At position 593, thymine was replaced with cytosine due to which valine was replaced by alanine at position 198 (593 T>C, p. V198A). Similarly, at position 648, the thymine was replaced by cytosine (648 T>C. Y216Y), at position 663, cytosine was replaced by thymine (663 C>T, Y221Y), and at position 666 the guanine was replaced by adenine (666 G>A, G222G). These changes have no effects on amino acid sequences and protein structures and were confirmed as silent mutations. In-vitro plant feeding assay revealed a significant difference in PBW mortality fed on Bt transgenic and non-transgenic cotton plants. In conclusion, our findings suggested good efficiency of Bt cotton while lacking any alarming mutation in cadherin and v-ATPase genes of local PBW isolate.
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