Cloning And Expression Analysis OfGhSOC1,GhFBO In Gossypium Hirsutum L.

Cotton is a major economic crop and an important strategic materials, so it will play an importance role in the development of national production. Since grain and corn production compete for the same land resource, there is a conflict in China with increasing population and decreasing arable land. With the wheat and cotton double cropping system, short-season cotton plays an important role in China. The MADS-box transcript factors can regulate the development of the flowering bud. In this paper, we isolate a MADS-box transcript factor named GhSOC1 and other gene GhFBO from a normalized cDNA library and analysis its expression patterns, at the same time, we also translated this gene to Arabidopsis to get more proof to demonstrate the funcation of this gene. The results are as follows:1.Total RNA and DNA were isolated from the leaves of CCRI 36. With RT-PCR and RACE, GhSOC1 was cloned. The full length of the gene is 1053 bp, including a 666 bp ORF, 101 bp 5'-UTR, 286 bp 3'-UTR, no introns.The startcode sequence was in conformity with the principle of Kozak and Phylogenetic analysis showed that the GhSOC1 was highly similarities with other SOC1 from different species, GhSOC1 protein has 221 amino acids. As a transcript factor, GhSOC1 protein locates in nuclear.2.By RT-PCR,I found that Gossypium arboretum L and Gossypium raimondiiL contain GhSOC1 gene. Therefore both A1 genome and D genome have this gene. The consequence show that this gene in different chromosome is nearly completely the same, the divergence between them is several nucleotide. If most of the genes between A1 and D chromosome is nearly the same,it may be difficult for us to assemble the fragment to one.3.The RT-PCR results showed that GhSOC1 expressed in every tissue. The expression was higher in the stems, but lower in the embryo. also detected the expression of GhSOC1 in different stage of SAM, the conclusion demonstrated that the highest expression stage is the third leaf flowed stage, which coincide with the result our lab got but the difference between the stages is conspicuous. 4. screening a cDNA library of CCRI 36, a new gene named GhFBO (Genebank number:JF498592)was isolated from CCRI36 which belonged to the TUB subfamily. The full-length of this gene is 1475bp and encodes a protein of 424 AA. The putative protein contains two conserved domain, F-box domain and Tubby C-terminal domain. Phylogenetic analysis displays GhFBO has the highest similarity with Malus x domestica TLP protein. Bioinformatics analysis indicats the protein is 47.6kd with PI 9.48 and has an signal peptide. The results of qRT-PCR demonstrated that the gene was expressed in all tested tissues with higher expression levels in flower, stem and leaf. The results indicate that this gene may be a homologous gene of MdTLP2 and plays an important role in the development of flower and morphogenesis.5.In order to understand the function more clear, we constract two overexpression vectors pBI121-GhSOC1 and pBI121-GhFBO as well as localized vector pBGFP-GhSOC1. We also got the the seeds of transgenitic Arabidopsis. GhSOC1 was localizated in nuclear.