Isolation And Expression Analysis Of A CaM And A FT Gene Involved In Flower Development From In Betula Platyphylla

Calmodulin is a key regulating protein in cells of organism, participating in and regulating enzyme activities and kinds of vital movements. Applying RT-PCR and RACE method, BplCaM gene was isolated from Betula platyphylla. Flowering Locus T (FT) gene and its homologues plays an important roles involved in promotion flowering plants and changes in developmental stage.The BplCaM gene with631bp in length contains450bp open reading frame and encodes a149-amino-acid polypeptide with a predicted molecular weight of16.85kDa and pI of4.11.The functional domains of its deduced amino acid sequence were confirmed as typical EFh motifs of CaM.The isolated gene was named as BplCaM, and registered in GenBank with accession number JN252700. Multiple sequences alignment and the phylogenetic tree analysis showed that it shared more than97%identity with other plant species. Expression of BplCaM was analyzed by the method of quantitative PCR normolized by act in gene of Betula. Results showed that BplCaM was more highly expressed in vegetative tissues than in reproductive tissues, and more higly in young organizations than in aging organizations. BplCaM expressed in multiple tissues with different levels. Furthermore, transcrip expression in a natural male inflorescense mutant was detected. BplCaM up-expressed in female inflorscences, young shoot and young leaves, indicating that it is associated with development of multiple tissues.The full cDNA of FT gene was isolated from Betula platyphylla Suk. by methods of reverse transcription polymerase chain reaction (RT-PCR) and5’and3’rapid amplification of cDNA ends (RACE). Results showed that FT gene contains an open reading frame (ORF) of525bp encoding174amino acids. Molecular weight of the deduced protein of BplFT is19.6kDa and the theoretical isoelectric point is7.73. The FT conserved protein domain of PEBP were confirmed in the sequence, so the isolated gene was named as BplFT, and registered in GenBank with accession number JQ409561. The deduced amino acid sequence shared74%-93%of identity with other sixteen plant species, the maximum identity with Ficus carica (93%) and minimum identity with Arabidopsis thaliana (74%). A phylogenetic tree was constructed according to multiple sequences alignment of all the seventeen plant species. Transcription expression of BplFT was analyzed by qRT-PCR in different tissues and periods in B. platyphylla. Results showed that BplFT was more highly expressed in vegetative organs than in floral organs, expression quantity more highly in mature tissues than in young tissues, the highest expression is mature stem.lt inferred that BplFT gene play an important role in mature tissues development and transfactor involved in the regulation of development of shoot meristematic tissues in Betula. In addition, a natural male inflorescence-abnormal mutant of B. platyphylla was used for transcription analysis of BplFT. Results showed that BplFT gene is expressed up-regulatedly in female inflorescences, male inflorescences, young leaves and young shoots, which predicted that BplFT should be not only be involved in the development of vegetative tissues, but also play some roles in the development of floral organs.