用实时定量PCR分析转roZABC基因积橙外源基因表达模式(摘要)

rolABC genes were cloned from T-DNA of Ri plasmid in Agrobactecium rhizogenes, apical dominance weakened, lateral branehes increased. internode shortened and some other special charaeters induced by transformation these genes into plants. It's very important to understand the molecular process of plant growth and development and charaeter formation, real一time PCR is high efficient and convenient method of analysis genes quantitative expression compare with Northern blotting. SYB Green I fluorescent dye used in this researeh, firstly, pDIN3514 vetor including rolABC genes transformed into Agrobacterium, and standard plasmids were isolated from Agrobacterium. Secondly, taking actin gene as reference gene, establishing two standard curves of rol genes and actin gene. At last, we studied on gene expression partern in different growth and development period, including dormancy, germination, leaf expansion and flowering stage by two Standard curves method and Z八ACT method. The resut reveals a Significant regularity of developmental trend of rol ABC genes expression in transgenic citrange, the expression intensity of rolA, rolBand rolC genes suceessively decreased, the relative expression value of rolC is 7一531 times and 6一198 times comPare to rol B and rolA respectively, forthermore, rol ABC genes expression in different transgenic 1ines also show obvious regularity, and rol ABC genes of E lines have highest expression in 3 transgenic lines. Combined with Southern blotting and transgenic citrange morphology observation，reasons of E lines dwarfing, deep green color and crimP of leaf and a Iarge number of lateral branehes comparing with B and D transgenic eitrange lines could connect with different insertion site of rol ABC genes and over-expression of rol ABC genes. However, The mechanisms of interaction between rol ABC genes and some closely related key dwarfing genes is hard to know at this moment, we make plan further this researeh in the next step.