Department of Plant Sciences, University of Hyderabad, Hyderabad, 500046, India.
Transgenic Res. 2012 Feb;21(1):193-200. doi: 10.1007/s11248-011-9521-y. Epub 2011 May 17.
Mungbean, Vigna radiata (L.) Wilczek is an important pulse crop that is widely cultivated in semi- arid tropics. The crop is attacked by various soil-borne pathogens like Rhizoctonia solani, which causes dry rot disease and seriously affects its productivity. Earlier we characterized the non-expressor of pathogenesis related gene-1(BjNPR1) of mustard, Brassica juncea, the counterpart of AtNPR1 of Arabidopsis thaliana. Here, we transformed mungbean with BjNPR1 via Agrobacterium tumefaciens. Because of the recalcitrant nature of mungbean, the effect of some factors like Agrobacterium tumefaciens strains (GV2260 and LBA4404), pH, L: -cysteine and tobacco leaf extract was tested in transformation. The transgenic status of 15 plants was confirmed by PCR using primers for nptII. The independent integration of T-DNA in transgenic plants was analyzed by Southern hybridization with an nptII probe and the expression of BjNPR1 was confirmed by RT-PCR. Some of the T(0) plants were selected for detached leaf anti-fungal bioassay using the fungus Rhizoctonia solani, which showed moderate to high level of resistance depending on the level of expression of BjNPR1. The seedling bioassay of transgenic T(2) plants indicated resistance against dry rot disease caused by R. solani.
绿豆,Vigna radiata(L.)Wilczek 是一种重要的豆类作物,广泛种植在半干旱热带地区。该作物受到各种土传病原体的攻击,如 Rhizoctonia solani,这会导致干腐病,严重影响其生产力。此前,我们对芥菜, Brassica juncea 的非致病相关基因 1(BjNPR1)进行了表征,它是拟南芥 AtNPR1 的对应物。在这里,我们通过根癌农杆菌将 BjNPR1 转化为绿豆。由于绿豆的顽固性,我们测试了一些因素的效果,如根癌农杆菌菌株(GV2260 和 LBA4404)、pH 值、L-半胱氨酸和烟草叶提取物。使用针对 nptII 的引物通过 PCR 确认了 15 株植物的转基因状态。通过 Southern 杂交用 nptII 探针分析了 T-DNA 在转基因植物中的独立整合,并用 RT-PCR 确认了 BjNPR1 的表达。根据 BjNPR1 的表达水平,选择了一些 T(0) 植株进行抗真菌生物测定,使用真菌 Rhizoctonia solani 进行离体叶片抗真菌生物测定,结果显示具有中度至高度抗性。对转基因 T(2) 植株的幼苗生物测定表明对 R. solani 引起的干腐病具有抗性。