Studies on the establishment of transformed apple-rootstock plants resistant to virus diseases by means of recombinant DNA technology

点击翻译按钮获取中文摘要

The present study was undertaken to establish the transformed apple-rootstock plants resistant to virus diseases by means of recombinant DNA technology. Using apple chlorotic leaf spot uirus (ACLSV) and a rootstock plant (Malus prunifolia) sensiive to ACLSV,following results are obtained.1. ACLSV was propagated in Chenopodium quinoa and succesfully purified. cDNAs were prepared from ACLSV-RNA extracted from purified virus and used to produce ACLSV-cDNA clones.2. The complete nucleotide sequence of ACLSV genome was determined by the dideoxynucleotide chain termination method. The genome is 7552 nucleotides excluding the poly (A) tail and contains three open reading frames (ORFs1,2, and3) .3. Coat protein gene (ORF3) was introduced to tobacco and Malus prunifolia plants by Agrobacterium Ti-mediated transformation.4.Western blot analysis indicated that ACLSV coat protein was expressed in transgenic tobacco plants and transgenic Malus prunifolia caluses. Tansformed Malus prunifolia plants are now growing.5.An antisense gene complementary to ORF2 region encoding 50K movement protein was transffered to tobacco and Malus prunifolia plants. PCR analysis indicated that the antisense gene was successfully introduced to tobacco plants. Transformed Malus prinofolia plants are now growing.6.To be summarized.The results mentioned above indicated that Ti-mediated trasnformation can be ued to introduce ACLSV genes to a rootstock plant. The method will be applied to produce transgenic plants resistant to other fruit tree virus diseases.