Betula Platyphylla Of The Male Inflorescence Developmental Differences In Gene Expression Analysis And Construction Of Plant Expression Vector Of Rab

Betula platyphylla male development cycle is longer, it’s nearly a year from the emergence of male inflorescence to pollen maturation, its early and mid development are the important periods to determine the male gametophyte development. In this paper, cDNA-AFLP molecular technique was used for sequence difference expression spectrum analyses of early and mid-term development male, received62TDFs totally,30TDFs of them have high homology with the genes in Genbank, the others with low homology are unknown sequences. The results of GO analysis illustrated that these known genes are mainly involved in metabolism process, cellular process, stimulate response, reproduction, signal transduction and biological control biological processes, their molecular functions are mainly related to catalytic activity, function of combination with molecular, enzyme regulator activity and the transfer activity, etc. We also found two important TDFs involved in reproduction and male flowers development, Bplbs658and Bplbsl99. In addition, we selected11TDFs which were used for qRT-PCR analysis of transcript expression of different tissues and male developmental processes. The results showed that these genes are not only involved in flower development, but also participated in the development of nutritive tissue, but they each has different tissue expression characteristics.Furthermore, we separated the cDNA of birch Rab gene by RT-PCR and expanded the full-length sequence by RACE technique, the total length of which is1017bp, with a612bp-length open reading frame, encoding203amino acids, named Bp1Rab. We translated the full-length sequence into amino acid sequence, predicted the protein structure of Rab gene and carried on the multiple sequence alignment, found that the amino acids sequence of the gene shared a similarity above90%with it in other plants. Through structure domain analysis we found that Rab protein has the conservative G domain structure of Rab family, then we forecasted the three-dimensional structure of Rab protein. Finally we builted a plant expression vector pROKII-Rab to lay a good foundation for later study in birch Rab gene function, agrobacterium mediated genetic transformation and infect arabidopsis and birch plants.