Cai Tiecheng, Chen Hua, Yan Liming, Zhang Chong, Deng Ye, Wu Shengxin, Yang Qiang, Pan Ronglong, Raza Ali, Chen Shunhui, Zhuang Weijian
Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Center of Legume Crop Genetics and Systems Biology/College of Agriculture, Oil Crops Research Institute, Fujian Agriculture and Forestry University (FAFU), Fuzhou, 350002, China.
State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.
Mol Biol Rep. 2022 Dec;49(12):11503-11514. doi: 10.1007/s11033-022-07817-z. Epub 2022 Sep 12.
Tobacco is an important economic crop, but the quality and yield have been severely impaired by bacterial wilt disease (BWD) caused by Ralstonia solanacearum.
Here, we describe a transgenic approach to prevent BWD in tobacco plants. A new root-specific promoter of an NtR12 gene was successfully cloned. The NtR12 promoter drove GUS reporter gene expression to a high level in roots but to less extent in stems, and no significant expression was detected in leaves. The Ribosome-inactivating proteins (RIP) gene from Momordica charantia was also cloned, and its ability to inhibit Ralstonia solanacearum was evaluated using RIP protein produced by the prokaryotic expression system. The RIP gene was constructed downstream of the NtR12 promoter and transformed into the tobacco cultivar "Cuibi No. 1" (CB-1), resulting in many descendants. The resistance against BWD was significantly improved in transgenic tobacco lines expressing NtR12::RIP.
This study confirms that the RIP gene confers resistance to BWD and the NtR12 as a new promoter for its specific expression in root and stem. Our findings pave a novel avenue for transgenic engineering to prevent the harmful impact of diseases and pests in roots and stems.
烟草是一种重要的经济作物,但其质量和产量受到青枯雷尔氏菌引起的青枯病严重损害。
在此,我们描述一种防止烟草植株发生青枯病的转基因方法。成功克隆了一个新的NtR12基因的根特异性启动子。NtR12启动子驱动GUS报告基因在根中高水平表达,但在茎中表达程度较低,在叶中未检测到明显表达。还克隆了来自苦瓜的核糖体失活蛋白(RIP)基因,并使用原核表达系统产生的RIP蛋白评估其抑制青枯雷尔氏菌的能力。将RIP基因构建在NtR12启动子下游并转化到烟草品种“翠碧一号”(CB - 1)中,产生了许多后代。在表达NtR12::RIP的转基因烟草品系中,对青枯病的抗性显著提高。
本研究证实RIP基因赋予对青枯病的抗性,且NtR12作为一个新的启动子使其在根和茎中特异性表达。我们的研究结果为转基因工程防止根和茎中病虫害的有害影响开辟了一条新途径。