不同水稻品种DEHY基因的转化研究及后代分析

发布时间：2018-04-25 19:03

本文选题：水稻 + 品种　；参考：《河北科技大学》2016年硕士论文

【摘要】：水稻作为三大主粮作物之一,占据着世界主要的土地种植面积,在我国尤其明显。随着转基因技术的出现与发展,水稻基因组的改造受到广泛重视,转基因技术被应用到高产、优质水稻的培育以及提高水稻抗旱、耐盐、抗虫和抗除草剂等方面。目前,水稻已成为转基因研究的一种模式作物。但是水稻种类繁多,在组织培养过程中,不同品种的组织培养特性以及植株再生能力存在较大差异;在基因工程改造过程中,不同品种转化效率也显著不同。本试验拟将6种不同水稻品种—鲲旱1号、旱稻2号、旱糯2号、旱糯3号、吉粳88和绥粳8号,利用农杆菌侵染法将DEHY基因转入其愈伤组织,研究不同水稻品种愈伤组织的表型、状态和增殖、再生能力以及转化效率的差异,并对转基因植株进行后代分析;通过除草剂筛选、PCR检测和Southern杂交,确定其拷贝数,选出单拷贝纯合体植株,为转基因技术在水稻育种应用中,提供有益参考。试验结果表明:(1)不同品种的水稻其愈伤组织的诱导率有所不同:旱糯3号愈伤组织的诱导率最高,达92.8%,而鲲旱1号愈伤组织的诱导率相对较低,仅81.6%;(2)不同品种的水稻其愈伤组织的表型和增殖能力区别较大,绥粳8号和吉粳88的生长状态更好一些;(3)不同品种的水稻分化能力有较大差别,绥粳8号和吉粳88分化能力相对较好,均能达到70%以上,而鲲旱1号分化能力最差,仅30.1%;(4)不同品种的水稻转化率有较大差异,最多相差28.9%,鲲旱1号转化率最低,仅3.18%。对获得水稻转基因植株后代进行分析,T1代经PCR鉴定,得到转基因阳性植株91株;对T2代转基因植株进行除草剂筛选,筛选出45个疑似转基因纯合体株系,经PCR鉴定,初步判定16个为纯合体株系,然后对其进行Southern杂交,选出2个单拷贝数株系。
[Abstract]:As one of the three main grain crops, rice occupies the main land area in the world, especially in China. With the emergence and development of transgenic technology, the transformation of rice genome has received extensive attention. The transgenic technology has been applied to high yield, high quality rice cultivation and improving rice drought resistance, salt tolerance, insect resistance and herbicide resistance. At present, rice has become a model crop in transgenic research. However, there are many varieties of rice, in the process of tissue culture, the different varieties of tissue culture characteristics and plant regeneration ability are quite different, in the process of genetic engineering, the transformation efficiency of different varieties is also significantly different. Six different rice varieties, Kun Han 1, Upland Rice 2, Hanuo 2, Hanuo 3, Jijing 88 and Suijing 8, were used to transfer DEHY gene into their calli by Agrobacterium tumefaciens. To study the difference of phenotype, status, proliferation, regeneration ability and transformation efficiency of callus of different rice varieties, and to analyze the progeny of transgenic plants, and to determine the copy number of transgenic plants by screening for herbicides by PCR detection and Southern hybridization. Single copy homozygous plants were selected to provide useful reference for the application of transgenic technology in rice breeding. The results showed that the callus induction rate of different rice varieties was different: the callus induction rate of Hanuo 3 was the highest, up to 92.8, while the callus induction rate of Kunhan 1 was relatively low. The phenotype and proliferative ability of callus of different rice varieties were different, and the growth state of Suijing 8 and Jijing 88 were better. The differentiation ability of Suijing 8 and Jijing 88 was better than 70%, but the differentiation ability of Kunhan 1 was the worst, only 30.1%.) the transformation rate of different rice varieties had great difference, the difference was 28.9% at the most, and Kuanghan 1 was the lowest, only 3.18%. 91 transgenic plants were identified by PCR, and 45 transgenic homozygous lines were screened by herbicide screening and identified by PCR. Sixteen homozygous lines were identified as homozygous lines. Two single copy lines were selected by Southern hybridization.
【学位授予单位】：河北科技大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S511

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