《根癌农杆菌介导转化AtNHX1基因甘蓝型油菜的研究》-毕业论文.docVIP

南京晓庄学院2012届本科毕业论文 PAGE 10 PAGE 2 南京晓庄学院本科生毕业论文 根癌农杆菌介导转化AtNHX1基因甘蓝型油菜的研究 Research of AtNHX1 Gene transformation in Brassica napus by Agrobacterium tumefaciens 所在系（院）：生物化工与环境工程学院 学 生 姓 名： 指 导 教 师： 研究起止日期：二○一一年三月至二○一二年五月 二○一二年五月 根癌农杆菌介导AtNHX1基因转化甘蓝型油菜的研究 摘要：以甘蓝型油菜带柄子叶为转化受体，通过根癌农杆菌介导法，将Na+/H+逆向转运蛋白AtNHX1基因转入甘蓝型油菜中。结果表明，外植体经浓度OD600为0.4的菌液中侵染8-10min，卡那抗性绿苗率达3.75℅，经过对抗性植株的PCR检测证明外源基因已经整合到油菜基因组中，为提高甘蓝型油菜的耐盐能力提供了新的途径。 关键词：根癌农杆菌；带柄子叶；卡那霉素；转基因甘蓝型油菜； Research of AtNHX1 Gene transformation in Brassica napus by Agrobacterium tumefaciens Student: LiangYi Tutor: Cai Xiao-ning Abstract： Chosen cotyledon petiole as the recipient for transformation. AtNHX1 gene was transferred into Brassica napus by Agrobacterium tumefaciens transferring system. The results showed that the best physiological and transforming condition was in OD600 0.4 liquid bacterium for 8-10 min, 3.75℅ kan-resistant green seedings were obtained. It was proved that NHX gene was inserted into the genome of oilseed by the methods of PCR.It could provide a new way to improve the salt tolerance about Brassica napus. Keywords：Agrobacterium tumefaciens;Cotyledons with petiole; Kanamycn；Transgenic Brassica napus; TOC \o 1-3 \h \u 目录 HYPERLINK \l _Toc13860 引 言 PAGEREF _Toc13860 4 HYPERLINK \l _Toc4474 1 实验材料 PAGEREF _Toc4474 5 HYPERLINK \l _Toc824 1.1 植物材料 PAGEREF _Toc824 5 HYPERLINK \l _Toc31659 1.2 菌种及质粒 PAGEREF _Toc31659 5 HYPERLINK \l _Toc24191 1.3 抗生素 PAGEREF _Toc24191 5 HYPERLINK \l _Toc29163 1.4 培养基的种类 PAGEREF _Toc29163 5 HYPERLINK \l _Toc25248 1.4.1植物培养基 PAGEREF _Toc25248 5 HYPERLINK \l _Toc13191 1.4.2细菌培养基 PAGEREF _Toc13191 5 HYPERLINK \l _Toc11246 1.5 农杆菌菌液的制备 PAGEREF _Toc11246 6 HYPERLINK \l _Toc17627 2 实验方法 PAGEREF _Toc17627 6 HYPERLINK \l _Toc26975 2.1 无菌苗的培养 PAGEREF _Toc26975 6 HYPERLINK \l _Toc3298 2.2 油菜基因型的筛选 PAGEREF _Toc3298 6 HYPERLINK \l _Toc4699 2.1.2预培养 PAGEREF _Toc4699 6 HYPERLINK \l _Toc2134 2.1.3分化培养 PAGEREF _Toc2134 6 HYPERLINK \l _Toc20449 2.1.4继代培养 P