The Study Of Screening And Expression Of The Genes Related To Flowering In The New-type Spring Brassica Napus

Brassica is one of major oil crops in the world,It is also a major source of edible oil in China with the first ranking of planting area and production in the world.Flowering time is a critical factor affecting the maturity period?yield and quality of Brassica.Therefore,it has important theoretical value and application prospect to study the genes related to flowering in Brassica.In this paper,the early-maturing rape?Brassica napus L.?86 were used as the material.Differentially expressed gene fragments in seeding stage and buding stage were screened by cDNA-AFLP technique,of which full-length cDNA sequences were cloned.Meanwhile,the expression pattern of the genes were analyzed in different tissues and growth periods by RT-qPCR.The main results were as following:?1?64 differentially expressed gene fragments in the seeding stage and buding stage of 86 rape were screened by cDNA-AFLP,in which 54 gene sequences were found in the rape genome,and 42 fragments of them were highly similar with known functional gene sequence of Arabidopsis.In addition,the 12 rest of gene sequences were considered as unknow because of no finding in any other plants although they were discovered in the rape genome.?2?The 3' end sequences of unknown gene 84-23?85-25?128-1?107-23?171-1?174-1 and the 5' end sequences of 128-1?107-23?167-12 were successfully cloned by RACE technology.The full-length cDNA sequences of 128-1 and 107-23 were amplified using the full-length primers designed according to 5' and 3' end sequences.The 128-1 gene was 1071 base pairs,with only one nucleotide synonymous mutation compared with the predicted gene XM₀13850601.1 in Brassica napus.The 128-1 gene encoded a protein containing 349 amino acid residues,and the protein might be related to the synthesis of TPR super-family proteins by amino acid homology comparison.The 107-23 gene was 1345 base pairs,of which the sequence was highly similar with the predicted gene XM₀13752386.1 in Brassica napus except for one mutant base in 5' end and two mutant base in 3' end.The 107-23 gene encoded a protein with 358 amino acid residues,which might be a transcription factor TCP2 related to flowering by amino acid homology comparison.?3?Through analysis the expression of gene 128-1 and 107-23 in different periods and tissues of 86 rape by RT-qPCR,it was obvious that the gene 128-1 and 107-23 were expressed in all growth period,however,the expression of flower tissue in florescence was the highest in 128-1 gene.It concluds that the differentially expressed gene 128-1 and 107-23 may be related to the flowering in Brassica napus,and the specific function needs to be genetically verified further.