本文選題：苯達松　切入點：CRISPR-Cas　出處：《基因組學與應用生物學》2017年08期

【摘要】：水稻CYP81A6基因(Bel)編碼的細胞色素P450單加氧酶對苯達松和磺酰脲類除草劑具有抗性,該基因的隱性純合突變體(bel)表現(xiàn)為苯達松敏感致死性,可作為植物化學致死標記,在水稻雜交制種中具有重要的應用價值。本研究在CYP81A6基因的首個外顯子中設計兩個靶位點,構(gòu)建了含多靶點的CRISPR-Cas9載體,通過根癌農(nóng)桿菌介導法轉(zhuǎn)化粳稻中花11,獲得了56株轉(zhuǎn)基因T0代陽性植株,提取其中9株基因組DNA進行靶位點測序檢測,發(fā)現(xiàn)基因敲除突變率為77.8%,其中純合突變率高達71.4%,對純合突變體水稻噴施苯達松,均表現(xiàn)出對苯達松敏感致死。為應用CRISPR-Cas9技術培育苯達松敏感致死水稻不育系和對其他水稻基因進行編輯提供理論依據(jù)。
[Abstract]:The cytochrome P450 monooxygenase encoded by CYP81A6 gene of rice is resistant to bentazone and sulfonylurea herbicides. The recessive homozygous mutant of this gene is bentazone sensitive to death and can be used as a chemical lethal marker in plants.It has important application value in hybrid seed production of rice.In this study, two targets were designed in the first exon of CYP81A6 gene, and a CRISPR-Cas9 vector containing multiple targets was constructed. By Agrobacterium tumefaciens mediated transformation of Japonica rice Zhonghua 11, 56 transgenic T0 positive plants were obtained.The gene knockout rate was 77.8%, and the homozygous mutation rate was 71.4%. Bentazon was sprayed on the homozygous mutant rice, all of which showed sensitivity to bentazone.It provides theoretical basis for the cultivation of Bentazon sensitive lethal rice sterile lines and the editing of other rice genes by using CRISPR-Cas9 technique.
【作者單位】： 南昌大學生命科學學院江西省分子生物學與基因工程重點實驗室;
【基金】：南昌大學“創(chuàng)新創(chuàng)業(yè)訓練計劃”項目、南昌大學研究生創(chuàng)新專項資金(cx2016193) 江西省研究生創(chuàng)新專項資金(YC2016-S063)共同資助
【分類號】：Q943.2
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本文編號：1725425