本文選題：苯達(dá)松　切入點(diǎn)：CRISPR-Cas　出處：《基因組學(xué)與應(yīng)用生物學(xué)》2017年08期

【摘要】：水稻CYP81A6基因(Bel)編碼的細(xì)胞色素P450單加氧酶對(duì)苯達(dá)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(dá)松敏感致死性,可作為植物化學(xué)致死標(biāo)記,在水稻雜交制種中具有重要的應(yīng)用價(jià)值。本研究在CYP81A6基因的首個(gè)外顯子中設(shè)計(jì)兩個(gè)靶位點(diǎn),構(gòu)建了含多靶點(diǎn)的CRISPR-Cas9載體,通過(guò)根癌農(nóng)桿菌介導(dǎo)法轉(zhuǎn)化粳稻中花11,獲得了56株轉(zhuǎn)基因T0代陽(yáng)性植株,提取其中9株基因組DNA進(jìn)行靶位點(diǎn)測(cè)序檢測(cè),發(fā)現(xiàn)基因敲除突變率為77.8%,其中純合突變率高達(dá)71.4%,對(duì)純合突變體水稻噴施苯達(dá)松,均表現(xiàn)出對(duì)苯達(dá)松敏感致死。為應(yīng)用CRISPR-Cas9技術(shù)培育苯達(dá)松敏感致死水稻不育系和對(duì)其他水稻基因進(jìn)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(xué)生命科學(xué)學(xué)院江西省分子生物學(xué)與基因工程重點(diǎn)實(shí)驗(yàn)室;
【基金】：南昌大學(xué)“創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃”項(xiàng)目、南昌大學(xué)研究生創(chuàng)新專項(xiàng)資金(cx2016193) 江西省研究生創(chuàng)新專項(xiàng)資金(YC2016-S063)共同資助
【分類號(hào)】：Q943.2
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本文編號(hào)：1725425