Cloning And Analyzing On LFY、NLY And TFL1 Genes Of Pinus Massoniana

Pinus massoniana is one of common timber tree species in southern China. Because of strong adaptability and fast growth, it has high economic and ecological value. However, few genes associated with flowering are reported. This dissertation cloned and analyzing FLO/LFY gene and TFL1 gene from P. massoniana, the results are as follows:FLO/LFY and its homologous genes are to control floral meristem.TFL1 and its homologous genes play important roles in maintaining vegetative growth and inflorescence meristem identity. Three full-length cDNA fragments of FLO/LFY and TFL1 were isolated from P. massoniana through RT-PCR and RACE, named them PmLFY, PmNLY and PmTFL1.Their sequences length are 1555 bp,1236bp and 854 bp,ORF of them are 1236 bp,933bp and 522 bp,encoding 411,310 and 522 amino acids respectively. Based on the three proteins of the physical and chemical analysis,the molecular formula of PmLFY is C2056H3258N602O618S20,the molecular weight is 46927.3,the PmNLY isC1028H2448N430O478S14,34939.6 and the PmTFL1 is C869H1362N242O250S8,194 56.3.The PmLFY and PmNLY proteins belong to FLO/FLY superfamily and the PmTFL1 protein belongs to PEBP superfamily. In gymnosperms, FLO/LFY gene is divided into LFY-like and NLY-like, PmLFY and PmNLY respectively belong to the two genes LFY-like and NLY-like group. The tissue specificity analysis show that PmLFY and PmNLY mainly expressed in bud, and and the expression in lateral bud is 1.8 and 1.3 times as high as that in terminal bud, compared to bud, the highest in its leaf PmTFL1 gene expression abundance, about 12.6,and the expression in shoots, twigs, roots were low.During construction of plant expression vector, by the enzyme digestion method to construct the expression vector, PmLFY and PmNLY are building the sense expression vector and PmTFL1 is antisense expression vector. Through the extraction DNA and RNA of transgenic tobacco, the PmLFY gene has been introduced into tobacco proved by PCR detection. Through the observing the phenotype of transgenic plants, the transgenic plants of plant height, leaf size is smaller than those no-transgenic, but not obvious. The transgenic plants flowered earlier than the no-transgenic.