Regeneration & Transformation System Of Cucumis Melo L. And The Full Length CDNA Cloning Of SPS Gene

Melon (Cucumis melo L.) is an important horticultural plant in gourd family (Cucurbitaceae), for its higher commodity and economic value. With the rapid development of plant cell and gene engineering technology, researchers are applying these technique to improve melon fruit quality, resistance and innovate on new germplasms. At present, the melon materials of tissue culture are almost hybrids and imported varieties.The rapidly reproduction of melon is needed in current production practice and the t echnology of tissue culture is just meeting these demands. Agrobacterium Tumefacie mediated MTV-CP,CMV-CP and TMV-CP genes transformed into melon had been repo rted successfully. It was hard to use hybrids in breeding because of its self-separationin its progeny, so we chose three different type of melon self-bred lines with low s ugar content to study the main factors that influence the regeneration and transformati on process. Moreover, we attempted pollen-tube pathway method to get the transgenic melon plants at frist time and set up the corresponding method of early transformantsselection. We cloned the full-length cDNA fragment of SPS, which is the key enzyme in sugar metabolism of melon fruit. The main results were as follows. 1 Systematic research on the effect factors of melon self-line regeneration Wu Kelan (thick skinned melon self-bred line ) and No.1 Lubo (thin skinned melon self-bred line) with low sugar content were tested. We studied individually the influential factors respectively, which were high quality aseptic seedlings, genotype of explant, age of seedlings, induction callus and adventitious buds medium, elongate an d rooting medium. Effect of 6-BA, ZT, KT, 2,4-D, GA, NAA, IAA, IB A, the p rimary phytohormones on melon regeneration and transformation , were also analysed and compared one to each other. We induced directly the adventitious buds from the explant of cotyledon petioles.The results indicated, the fact that high quality of seedlings could been obtained by the germination of sterilized seed; The cotyledon petioles of 6d Wu Kelan and 4 d No.l Lubo seedling were favourable to induce callus and buds; The optimum bud medium of Wu Kelan and No.l Lubo were MS+ 1.0mg/L6-BA+ 3.0-3.5mg/L ZT+ 0.1...