Cloning And Expression Analysis Of Hmologous FT And LEAFY In Allium Cepa L

Onion is liliaceous of allium herbs and propagates by seed,which was green vernalization plant in developmental process. Low temperature and photoperiod are the necessary conditions for bolting and flowering.It is reported that FT (FLOWERING LOCUS T) belongs to a"cross talk"during flowering development, which is the key gene determing flowering time located in photoperiod. LEAFY controlled the floral meristem shifting from inflorescence meristem and regulating flower time and inflorescence morphogenesis. Until now, onion homologous FT and LEAFY genes related have not been reported.In this study, Onion 1007 strain as the material,whose leaves were used to investigate the method of extracting RNA . Two pairs of degenerate primers were designed according to the conservative regions of the plant floral meristem identity gene-LEAFY and FT homologousgenes.RNA from Leaves during the vegetative growth and early bolting stem apical meristem as template, respectively.Cloned FT and LEAFY homologous genes by RT-PCR and analyzed spatial and temporal patterns of gene expression preliminarily.Lay the foundation for the study on the molecular mechanism of bulb crops bolting and flowering.Experimental results were as follows:1The performances of five extraction methods of total RNA: Trizol,modified Trizol,SDS,modified SDS and TIANDZ Column Plant RNAout ,were compared with in this report.The TIANDZ Column Plant RNAout was the best method of all. It's simple to operate and contains high-quality and highly-complete RNA .It was indicated that the TIANDZ Column Plant RNAout extraction could be worth further biological research..2.Designed the primers from the conserved sequence of FT hmologous and used RT - PCR technology, we had first cloned FT homologous gene,which was a CDS sequences contained 185bp and coded 62 amino acids.3.Designed the primers from the conserved sequence of LEAFY hmologous and used RT - PCR technology, we had first cloned LEAFY homologous gene,which was a CDS sequences contained 324bp and coded 108 amino acids.4.FT had no expression on leaves in its seedling stage and gradually more expressions in its vegetative stage, with the occurrence of reproductive growth, it has lower expression in pseudostems till flowing period which has no expressions.5.LFY had no expression on leaves in its seedling stage and vegetative stage, with the occurrence of reproductive growth, it has large expressions in the apical meristems.During flowing period,LFY had no expressions.