Study Of Betula Platyphylla Floral-timing Genes

This research is concerned to clone the flower-timing genes of Betula platyphalla under the intensive condition by orthologus gene cloning technique. The purpose is to reveal the control mechanism of early flowering genes of this species, and also to analyze some special gene's expression in that condition.The best time to collect the materials for cloning is discussed in this paper, which is from the end of May to the beginning of June each year. It means there is only a week for the materials to transit from apex dividing tissue to flower inflorescence, but still a little change with temperature every year.Since there are more things in the bud tissue of the male flower, such as phenolic compounds, polysacchrides, proteins and some other secondary metabolites, three methods used to isolate RNA are tested in this assay, which are the enzyme digesting methods of CTAB, CTAB-DNA and SDS-DNA. At last, CTAB-DNA and SDS-DNA methods are used in this experiment.On the basis of optimizing experiment procedure and PCR system of the materials, the initial reversion transcription system and orthologus gene cloning technique are established.Seven pairs of degeneracy primers for cloning are schemed out by using Bioinformatics software. After selection, one pair of primer (No.l: PS 5'-ARAARGCNCAYGARAT , PA 5' -RCATCCANGGNGGCAT) is used to do PCR amplification in the experiment.Three special bands as PCR products, obtained by using primer No.l, are reclaimed, and then used to cloning and sequencing. Through homologous comparison with data from International Nucleotide Databank, the band's functions have been simply analyzed. It shows that the longest band (675bp) has lower homology with flower-timing genes, but there is 80 percentage of homology with pectinesterase during Citrus development, in 184 score of HSP. It is clear to see a functional domain by modeling three-dimension structure of the protein. Hereby, it may probably concern this band as a partial segment of pectinesterase of Betula platyphalla. The other study will be done in the future.