In silico analysis cloning and overexpression of miR172 in potato (Solanum tuberosum L. SSP andigena)

Abstract

MicroRNA172 (miR172), one of the most well studied microRNAs (miRNAs) in plants, mediates cleavage of APETALLA2 (AP2)/APETALLA2 like (AP2 like) mRNAs. AP2/AP2 like genes are involved in fruit ripening, meristem identity, flowering and many other functions in plants. In addition, AP2 like genes are also reported to be involved in stress response. In potato (Solanum tuberosum), only one AP2 like gene, called RAP1 is reported so far. RAP1 inhibits tuberization in potato by repressing StBEL5, the mobile signal for tuberization. miR172 known to enhance tuberization mechanism by down-regulating its target RAP1. In this study, a detail In analysis of eight putative miR172 target genes has been performed using various softwares and databases. Our results suggest that four of them are AP2 like genes, one has a b-Zip transcription factor domain, one codes for BTF3 like transcription factor whereas the rest two are coding for proteins having unknown functions. Unigene sequences for seven of them were also retrieved. All the predicted target mRNAs of miR172 show transcript cleavage as their mode of inhibition and have one miR172 binding site. In order to understand the miR172 and target gene regulation, 35S::miR172 overexpression transgenic potato plants have been generated and putative clones have been validated by RT-PCR analysis. Further experiments are in progress.