Cloning, characterization and expression analysis of <em>APETALA2</em> genes of <em>Brassica juncea</em> (L.) Czern.

604-610The APETALA2/Ethylene-Responsive Factor (AP2/ERF) is one of the largest gene families encoding several plant specific transcription factors. It plays significant roles in growth and development process, biotic and abiotic stresses, and responses to hormones. AP2 is a homeotic gene governing floral meristem specification, floral organ determination and floral homeotic gene expression in Arabidopsis. The basic structure of AP2 gene was unchanged during evolution in diploid species. The present study was undertaken to find whether AP2 has undergone any change in structure or expression pattern during evolution of allopolyploid Brassica juncea. We cloned AP2 orthologs and c-DNAs from B. juncea and B. nigra. B. juncea was found to carry three AtAP2 orthologs. Comparison of BjAP2 genes with AP2 orthologs from progenitor species, B. rapa and B. nigra showed that two of the BjAP2 genes were derived from B. rapa and one from B. nigra. BjAP2 genes have retained its characteristic AP2 domain and miR172 complementary sequences. mRNAs originated from three AP2 orthologs were detected in all the tissues examined, namely, leaf, flower buds and seedling, indicating absence of sub-functionalization of AP2 during polyploid evolution. However, one of the B. rapa copies gave alternatively spliced AP2 transcript which lacked the second exon. Consequently, the splice variant could not be translated into functional AP2 protein. Considering that miR172 suppresses translation of AP2 transcripts, the alternatively spliced transcript could still play important regulatory role by limiting the availability of miR172 molecules to bind to functional AP2 transcripts. qRT-PCR analysis of BjAP2 expression in different accessions of B. juncea with contrasting seed size indicated that BjAP2 is not a major determinant of seed size in mustard