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通过表达核糖体失活蛋白α-苦瓜素增强水稻(Oryza sativa L.)对炭疽病的抗性。

Enhanced resistance to blast fungus in rice (Oryza sativa L.) by expressing the ribosome-inactivating protein α-momorcharin.

机构信息

State Key Laboratory of Hybrid Rice, Department of Genetics, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, People's Republic of China.

State Key Laboratory of Hybrid Rice, Department of Genetics, College of Life Sciences, Wuhan University, Wuhan 430072, Hubei Province, People's Republic of China.

出版信息

Plant Sci. 2014 Mar;217-218:1-7. doi: 10.1016/j.plantsci.2013.11.012. Epub 2013 Nov 28.

Abstract

Rice blast caused by Magnaporthe grisea is one of the three major diseases that seriously affect the rice production. Alpha-momorcharin (α-MC), a ribosome-inactivating protein (RIP) isolated from Momordica charantia seeds, has antifungal effects in vitro. In this study, the α-MC gene was constitutively expressed under the control of the 2×35S promoter in transgenic rice (Oryza sativa L.) using an Agrobacterium tumefaciens-mediated method. The nine transgenic plants were obtained and confirmed by PCR and RT-PCR, and the four (B2, B4, B7 and B9) of them whose copy numbers were 1, 2, 3 and 3, respectively, were shown to express the α-MC protein by Western blot. The molecular weight of α-MC in transgenic plants was approximately 38 kDa larger than the purified α-MC protein (28 kDa) in vitro. When the confirmed T1 generations were inoculated with a suspension of M. grisea spores for ten days, the lesions on leaves of transgenic plants were much lesser than those found on wild type (WT). According to the criteria of International Rice Research Institute standard, the mean values for morbidity and disease index numbers were 29.8% and 14.9%, respectively, which were lower than for WT. It is unclear whether RIPs could impact plant fitness and however our results suggest that the α-MC protein is an effective antifungal protein preventing rice blast in transgenic rice.

摘要

稻瘟病由稻瘟病菌引起,是严重影响水稻生产的三大病害之一。α-苦瓜核糖体失活蛋白(α-MC)是从苦瓜种子中分离得到的一种核糖体失活蛋白,具有体外抗真菌作用。本研究采用根癌农杆菌介导法,将α-MC 基因置于双 35S 启动子的控制下,在转基因水稻(Oryza sativa L.)中组成型表达。通过 PCR 和 RT-PCR 获得了 9 株转基因植株,并得到了证实,其中 4 株(B2、B4、B7 和 B9)的拷贝数分别为 1、2、3 和 3,通过 Western blot 显示表达了 α-MC 蛋白。转基因植株中 α-MC 的分子量比体外纯化的 α-MC 蛋白(28 kDa)大约大 38 kDa。当确认的 T1 代用稻瘟病菌孢子悬浮液接种 10 天后,转基因植株叶片上的病斑明显少于野生型(WT)。根据国际水稻研究所标准的标准,发病率和病情指数的平均值分别为 29.8%和 14.9%,均低于 WT。目前尚不清楚核糖体失活蛋白是否会影响植物的适应性,但我们的结果表明,α-MC 蛋白是一种有效的抗真菌蛋白,可防止转基因水稻发生稻瘟病。

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