PCR based site-directed mutagenesis: An extensively used tool for the functional genomics studies in begomoviruses

Site-directed mutagenesis (SDM) is an important tool that allows the quick and efficient manipulations of specific residues in DNA sequences for investigating their importance in structure and function of proteins. Out of several techniques available for this purpose, SDM coupled with PCR are widely used because of its simplicity and effectiveness, and are less time consuming. The technique uses pairs of oligonucleotide primers carrying the desired mutation for introducing specific changes in the target DNA sequence. Begomoviruses alone, or in combination with satellite molecules pose significant threat to global agriculture for their worldwide occurrence on a broad range of crops. Understanding the functions of genes available on the begomoviral genome and satellite molecules is the key for devising effective disease control strategies. PCR-based SDM has proven to be instrumental in unravelling the functions of genes and molecular mechanism of disease development by understanding the dynamic interplay between begomoviruses and the host plants. This has paved a way for the development of innovative antiviral interventions and sustainable agricultural practices to mitigate the impact of these economically important plant pathogens on global food security and crop production. This review briefly summarized about the overall concept and methods of SDM with emphasis on PCR based SDM and its applications so far in the functional genomics studies of begomoviruses.