Cloning And Analysis Of CO And FT Gene In Anthurium Andraeanum

Anthurium andraeanum is popular potted plant and cut flower in the world, for its high ornamental value. At present, China has become a major global productior and market. But its growthperiod is much longer (potted flower is two years), and its blossom is vulnerable to poor nutrition,temperature and illumination conditions. It has thepositive significanceshortening the period and increasing resistance and the production of cut flower by parsing flowering molecular mechanism and regulating the expression of important genes.Plant flowering is regulated by the complex molecular network, mainly including vernalization pathway, gibberellin pathway, autonomous regulation pathway, and photoperiodic pathway. FT and CO gene as the integration of a variety of signal factors, play an important role intansmitting flowering signals of upstream and regulatinng the expression of flowering gene of downstream. This study is on cloning, identification and bioinformatics analyses of A. andraeanum FT and CO genes. The results are as follows:(1) Based on the transcriptome mining and blast analysis, anthurium PEBP family at least contains five members and one (AaMFT1) belongs to the MFT sub-family, three (AaFTl, AaFT2, AaFT3) to the FT sub-family, and the other one (AaFTL1) to the TFL sub-family.(2) Via the transcriptome characterion and RACE amplification, two CO like genes were cloned and named AaCO1, AaCO2. AaCO1included a1263bp of ORF, coding420amino acids. AaCO2contains a1257bp of ORF, coding418amino acids. Both produced proteins contained B-box structure on N termianl and CCT motif on C terminal, with strictly conservative sequence. Phylogenetic analysis revealed that AaCO1, AaCO2and Arabidopsis thaliana CO family III belong to a group, and AaCO1and AaCO2are close to A. thaliana COL9and COL10.(3) qRT-PCR analysis showed that FT-like genes had different expressing characteristics in various organizations. AaMFT expressed highest in leaf, root second, stem the lowest. AaTFL expressed highest in root, lowest in others and leaf didn’t express. AaFTl expressed highest in spadix quantity, root second, stem the lowest. AaFT2expressed highest in spadix, root second, petioles the lowest and AaFT3expressed highest in root, petioles second, spathe the lowest, and leaf didn’t express. Under the condition of long day (16h/8h), AaMFT from leaves was highest in16h, and AaTFL, AaFTl, AaFT2and AaFT3from leaves were highest in12h. Under the conditions of short day (8h/16h), AaMFT and AaFTl were highest in12h, AaFT2was highest in16h, AaFT3was highest in4h and AaTFL was highest in20h. (4) Expression law of AaCO1, AaCO2gene was similar, root highest, stem lowest. Under the condition of long day (16h/8h), both were highest in16h, and under the condition of short day, both were highest in12h.(5) AaFT1genome sequence contains four exons and three introns. Subcellular localization showed that AaFTl had no obvious distribution in the cell. Genotype expression vector p35S::FT of AaFTl, bud meristem expression vector pFD::FT and sieve tube specific expression vector pSUC2::FT had been builtand turned to root carcinoma agrobacterium to prepare for plant expression.Results had positive significance in further validation of FT and CO gene function, analytical molecular mechanism of A. andraeanum flowering signal, and subsequent flowering regulation and so on.