Isolation And Functional Analysis Of Flower-related Genes FVE And EFS1 Of Arabidopsis Thaliana

A late-flowering mutant 221-1 and an early-flowering mutant 426-2 were obtained from T-DNA mutational seeds in Columbia(Col-0)genetic background,which is single copy of exogenetic DNA in the genome of the Arabidopsis thaliana.The late-flowering mutant caused a pleiotropic phenotype including big and thick leaves,increased numbers of rosette leaves,prolonged vegetative growth,delayed flowering time(60 days);the early-flowering mutant caused a pleiotropic phenotype in short photoperiod including small leaves,decreased numbers of rosette leaves,increased numbers of branches and cauline leaves,advanced flowering time(10 days),but the early-flowering mutant had the same phenotype with wild type in long photoperiod.We had optimized the system and the factors that affected the efficiency of TAIL-PCR. The flanking sequence of T-DNA in the mutant 221-1 was obtained by optimized TAIL-PCR.It was known that the locus of the insert was in the sixth extron of AT2G19520.1(FVE gene)by BLAST on TAIR(The Arabidopsis Information Resource) and we can be sure that FVE is an important gene in autonomous flowering pathway.The mutant 221-1 is the similar with fve-3 from Columbia but is allelic mutant of fve-1 and fve-2 from Landsberg erecta.So the mutant 221-1 was named fve-4.Compared with the other mutants of FVE,fve-1,fve-2 and fve-3,fve-4 obtained in this study displays different locus which is broken and later flowering time.The loci which are broken of fve-1,fve-2, fve-3 and fve-4 are the seventh extron,the eighth extron,the first extron,the sixth extron, respectively.Compared with the wild type,the flowering time of fve-1,fve-2 and fve-4 are delayed 12 days,20 days,60 days,respectively.The flanking sequence of T-DNA in the mutant 426-2 was obtained by Genomic Walking.Through BLAST analysis,we are sure that the locus of the insert is in the 5'-NCR(noncoding region)of AT4G36680.1,named EFS1.It encodes pentatricopeptide (PPR)repeat-containing protein,is also considered to be a kind of prenyltransferase.It isn't the report that EFS1 participate in flowering pathways at present,the mutant 426-2 is named early flowering in short days 1(efs1).To determine the roles of EFS1 in flowering pathways,some important genes related to flowering time were investigated in the efs1 mutant by RT-PCR.We can be sure that a few characteristic traits related to flowering of the efs1 mutant is the similar to early-flowering mutant in photoperiod-dependent flowering pathway.The function of EFS1 is the similar to EARLY BLOTING IN SHORT DAYS(EBS)which restrains the express of FT.Therefore,EFS1 may be a new member of Photoperiod-dependent flowering pathway,which plays the function through promote the express of EMF and TFL1 which restrains the flowering.To further determine the function of EFS1,T-DNA Plant Expression Vector of the EFS1 gene,named pCAMBIA1300-EFS1(and promoter)-NOS.