拟南芥磺酰脲类除草剂抗性基因作为一种新的可选择标记用于生产可育的转基因水稻植株。
A Sulfonylurea Herbicide Resistance Gene from Arabidopsis thaliana as a New Selectable Marker for Production of Fertile Transgenic Rice Plants.
机构信息
Department of Plant Pathology and Crop Physiology, Louisiana State University, Baton Rouge, Louisiana 70803-1720.
出版信息
Plant Physiol. 1992 Oct;100(2):662-8. doi: 10.1104/pp.100.2.662.
A mutant acetolactate synthase (ALS) gene, csr1-1, isolated from sulfonylurea herbicide-resistant Arabidopsis thaliana, was placed under control of a cauliflower mosaic virus 35S promoter (35S). Rice protoplasts were transformed with the 35S/ALS chimeric gene and regenerated into fertile transgenic rice (Oryza sativa) plants. The 35S/ALS gene was expressed effectively as demonstrated by northern blot hybridization analysis, and conferred to transformed calli at least 200-fold greater chlorsulfuron resistance than nontransformed control calli. Effective selection of 35S/ALS-transformed protoplasts was achieved at extremely low chlorsulfuron concentrations of 10 nm. The results demonstrated that the 35S/ALS gene is an alternative selectable marker for rice protoplast transformation and fertile transgenic rice production. The results also suggest that the mutant form of Arabidopsis ALS enzyme operates normally in rice cells. Thus, the mechanism of protein transport to chloroplast and ALS inhibition by chlorsulfuron is apparently conserved among plant species as diverse as Arabidopsis (dicotyledon) and rice (monocotyledon).
从抗磺酰脲类除草剂的拟南芥中分离得到突变的乙酰乳酸合酶（ALS）基因 csr1-1，该基因受花椰菜花叶病毒 35S 启动子（35S）的控制。用 35S/ALS 嵌合基因转化水稻原生质体，并再生出可育的转基因水稻（Oryza sativa）植株。Northern blot 杂交分析表明，35S/ALS 基因得到了有效表达，并赋予转化的愈伤组织比非转化对照愈伤组织至少 200 倍更高的氯磺隆抗性。在非常低的氯磺隆浓度（10 nM）下，实现了 35S/ALS 转化原生质体的有效选择。结果表明，35S/ALS 基因是水稻原生质体转化和可育转基因水稻生产的替代选择标记。结果还表明，拟南芥 ALS 酶的突变形式在水稻细胞中正常运作。因此，蛋白向叶绿体的转运机制和氯磺隆对 ALS 的抑制作用在拟南芥（双子叶植物）和水稻（单子叶植物）等不同植物物种中显然是保守的。
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