本文選題：水稻 + 品種　； 參考：《河北科技大學(xué)》2016年碩士論文

【摘要】：水稻作為三大主糧作物之一,占據(jù)著世界主要的土地種植面積,在我國尤其明顯。隨著轉(zhuǎn)基因技術(shù)的出現(xiàn)與發(fā)展,水稻基因組的改造受到廣泛重視,轉(zhuǎn)基因技術(shù)被應(yīng)用到高產(chǎn)、優(yōu)質(zhì)水稻的培育以及提高水稻抗旱、耐鹽、抗蟲和抗除草劑等方面。目前,水稻已成為轉(zhuǎn)基因研究的一種模式作物。但是水稻種類繁多,在組織培養(yǎng)過程中,不同品種的組織培養(yǎng)特性以及植株再生能力存在較大差異;在基因工程改造過程中,不同品種轉(zhuǎn)化效率也顯著不同。本試驗擬將6種不同水稻品種—鯤旱1號、旱稻2號、旱糯2號、旱糯3號、吉粳88和綏粳8號,利用農(nóng)桿菌侵染法將DEHY基因轉(zhuǎn)入其愈傷組織,研究不同水稻品種愈傷組織的表型、狀態(tài)和增殖、再生能力以及轉(zhuǎn)化效率的差異,并對轉(zhuǎn)基因植株進行后代分析;通過除草劑篩選、PCR檢測和Southern雜交,確定其拷貝數(shù),選出單拷貝純合體植株,為轉(zhuǎn)基因技術(shù)在水稻育種應(yīng)用中,提供有益參考。試驗結(jié)果表明:(1)不同品種的水稻其愈傷組織的誘導(dǎo)率有所不同:旱糯3號愈傷組織的誘導(dǎo)率最高,達92.8%,而鯤旱1號愈傷組織的誘導(dǎo)率相對較低,僅81.6%;(2)不同品種的水稻其愈傷組織的表型和增殖能力區(qū)別較大,綏粳8號和吉粳88的生長狀態(tài)更好一些;(3)不同品種的水稻分化能力有較大差別,綏粳8號和吉粳88分化能力相對較好,均能達到70%以上,而鯤旱1號分化能力最差,僅30.1%;(4)不同品種的水稻轉(zhuǎn)化率有較大差異,最多相差28.9%,鯤旱1號轉(zhuǎn)化率最低,僅3.18%。對獲得水稻轉(zhuǎn)基因植株后代進行分析,T1代經(jīng)PCR鑒定,得到轉(zhuǎn)基因陽性植株91株;對T2代轉(zhuǎn)基因植株進行除草劑篩選,篩選出45個疑似轉(zhuǎn)基因純合體株系,經(jīng)PCR鑒定,初步判定16個為純合體株系,然后對其進行Southern雜交,選出2個單拷貝數(shù)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.
【學(xué)位授予單位】：河北科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S511

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1 閆素紅;不同水稻品種DEHY基因的轉(zhuǎn)化研究及后代分析[D];河北科技大學(xué);2016年

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