農(nóng)桿菌介導(dǎo)轉(zhuǎn)錄因子CBF轉(zhuǎn)化甘藍(lán)型油菜

生物工程論文編號(hào):SW108  論文字?jǐn)?shù):10758,頁(yè)數(shù):24

 目  錄
中文摘要 I
英文摘要 II
目錄 III
1. 緒論 1
 1.1 冷馴化及CBF家族概述 1
 1.2 CBF家族與植物抗凍性 3
2. 材料與方法 7
 2.1 材料 7
 2.2 方法 8
3. 結(jié)果與分析 12
 3.1 轉(zhuǎn)基因油菜植株的獲得 12
 3.2 轉(zhuǎn)化植株P(guān)CR檢測(cè) 13
 3.3 轉(zhuǎn)化植株Southern雜交分析 13
4. 總結(jié)與展望 15
致謝 16
參考文獻(xiàn) 17

摘 要：油菜是我國(guó)最主要的油料作物，有甘藍(lán)型油菜、白菜型油菜和芥菜型油菜等9個(gè)種，其中許多是具有重要經(jīng)濟(jì)價(jià)值的油料與蔬菜作物。但是當(dāng)環(huán)境溫度低于10℃～20℃時(shí)油菜就會(huì)受到傷害，影響它的正常生長(zhǎng)發(fā)育，嚴(yán)重的甚至造成死亡。目前很多作物在不同程度上都受到低溫的影響，低溫這一重要的環(huán)境因子正嚴(yán)重影響油菜的生長(zhǎng)、發(fā)展、生存及分布。因此，針對(duì)耐逆性（抗寒性）相關(guān)的轉(zhuǎn)基因油菜的研究，本試驗(yàn)以成熟油菜“超油2號(hào)”種子培養(yǎng)出的下胚軸愈傷組織為受體材料，運(yùn)用農(nóng)桿菌介導(dǎo)法將轉(zhuǎn)錄因子CBF基因?qū)�入供試品種的基因組中，得到的油菜植株經(jīng)PCR檢測(cè)，Southern雜交驗(yàn)證分析，獲得轉(zhuǎn)基因植株60株，轉(zhuǎn)化頻率為1.2%。已轉(zhuǎn)入目的基因植株的耐逆性的鑒定正在準(zhǔn)備之中。
關(guān)鍵詞：CBF；根瘤農(nóng)桿菌；PCR；Southern雜交

Agrobacterium tumefaciens-Mediated Transformation of Brassica napus L. with CBF Gene
Abstract: Rape is the most important oilseed crop, there are 9 kinds of rapes, Brassica napus, he cabbage rape, the leaf mustard rape and so on. Many of them have the important economic values. But when the environment temperature is lower than 10℃~ 20℃ rape may be injured, its normal growth would be affected, even worse, it would be died. At present, many crops affected by low temperature in the varying degree. But the low temperature of this important environmental factor that is seriously affecting rape's growth, development, survival and distribution. In this study, a gene for CBF was incorporated into genome of "super-oil No.2", a new Brassica napus L. strain with super-high oil content, by Agrobacterium tumefaciens-mediated transformation with hypocotyl segments as recipient materials. Based on PCR assay and Southern hybridization analysis, 60 transgenic plants were obtained and the transformation frequency was 1.2%. The identification of transgenic plants is being prepared.
Keywords : CBF; Agrobacterium tumefaciens ; PCR; Southern hybridization