农杆菌介导抗虫基因ppa的旱稻转化-植物学专业毕呢业论文.docxVIP

河北科技大学硕士学位论文 河北科技大学硕士学位论文 万方数据 万方数据 万方数据 万方数据 II AbstractAbstract Abstract Abstract Due to the shortage of water resource，rice production is restricted．So upland rice as a less water and drought resistant variety has good development and application prospects． As pests is an important factor to decrease the yield of rice，the use of transgenosis to cultivate the upland rice with insect resistance，has economic benefits and ecological benefits．尸剧gene has insecticidal physiological activity,whose application has a bright prospect in insect-resistant transgenic crop breeding． This research chose“Handao 2”as the experimental material，with Bar gene as biomarker for screening，and then transformed the insect—resistant gene of P剧by Agrobacterium-mediated transformation．Finally，this study optimized the conversion system，and produced the transgenic upland rice plants，who has the herbicide resistance and the activity of insect-resistant． To get T1 generation plants which have resistance of herbicide，0．5 mg／mL of the glufosinate was spray in field．In the laboratory,T2 generation transgenic lines were homozygous screening，lines with herbicide resistance were identified by PCR to find homozygous lines． In this study．Handa02 with PPA gene positive strain was identified by Southern hybridization．There are many factors that effect Southern hybridization，such as the digestion system，DNA concentration of the sample，concentration of the probe， hybridization conditions．The results showed that，adding 1 5 gg of DNA，90U endonuclease to the 60 gL digestion system and digesting 1 5-1 6 h is the most suitable． Adding 1 50 ng of probe to 5 mL hybridization solution in the hybridization system is most suitable．Southern blot analysis showed that．P以gene integrated into the genome of Handa02 in the form of multiple copies Key words Upland rice；Agrobacterium—mediated；Herbicide；PCR；Southern blot III 河北科技大学硕士学位论文 河北科技大学硕士学位论文 万 万方数据 IV 目 目 录 目 录 摘 要 ．．I Abstract ． ．．． ． ． ． ． III 第1章绪 论 l 1．1植物转基因技术 1