Identification of two embryo genesis-related genes in sweet pepper (Capsicum annuum L.) genome

Plant tissue culture techniques are essential part of modern plant biotechnology. Among them especially valuable are methods for production of haploid plants via anther cultures because they possess single copy of each gene and its functions can be easily manipulated and observed in the phenotype. Capsicum annuum (L.) produces sporadically successful anther cultures. Some cultivars produce embryos readily, while others do not respond at all. Many authors attributed variations in ability of plants to produce anther culture to differences in their genotype. It was also reported that the switch of microspores developmental programme from garnetophyte- to sporophyte-type is regulated by transcription factors like BABY BOOM (BBM) and LEAFY COTYLEDON (LEC). Genes encoding such transcriptional factors have not been reported in sweet pepper yet. We used annotated in NCBI sequences of BBM and LEC to design degenerative primers for highly conservative regions in these genes and used them to search by PCR reactions for BBM- and LEC-like sequences in the genome of Capsicum annuum (L.), cultivar Stryama. The isolated PCR fragments were cloned and sequenced. The nblast in NCBI database revealed high similarity between our sequences and annotated BBM and LEC sequences. Next the obtained sequences were subjected to bioinformatics analyses to determine the exon-intron structure. Exons were translated in-silico in protein products. The protein blast of our BBM in NCBI revealed the presence of AP2-type functional domain, typical of BBM gene family. The LEC sequence carries CBFDNFYB-HMF motive, typical of LEAFY COTYLEDON gene family. This is the known report of existence of BBM-like and LEC-like genes in Capsicum annuum (L.) genorne.