The Study Of The RT-PCR Detection Systems On Apple Mosaic Virus (ApMV) And Prunue Dwarf Virus (PDV)

The fruit trees viruses are threatening growth and development of fruit tree seriously.After the several factors that affected RT-PCR were examined ,the RT-PCR detection systems for ApMV and PDV were established.The RT-PCR detection systems for ApMV were optimized.The cDNA sequences of ApMV and PDV were analysis. The main research results were follows:The RT-PCR detection system for ApMV and PDV was established. The highly efficient RT-PCR detection systems of ApMV were optimized.The proper concentration of dNTPs and TaqE was 100μmol/L and 0.8 U,respectivly.The concentration of complementary primer and homologious primer should be 0.18μmol/L－1.8μmol/L and 0.412μmol/L－4.12μmol/L. The concentration of MgCL2 was up to 0.2 mmol/L.Reannealing time was 60 senconds and Reannealing temperature was between 48℃ and 56℃.Denaturation time was up to 45 senconds and Denaturation temperature was 92℃.Extention time was 60 senconds. The fragments of the partial CPgene of ApMV and PDV were obtained by RT-PCR detection system and were cloned to the pUCmT vectors successfully.Then,the inserted fragments were sequenced.The homologies of DNA sequence were up to 92.17% and 99% compared with the published ones.