National Research Centre on Plant Biotechnology, Pusa Campus, New Delhi, 110012, India.
School of Agriculture and Food Sciences, The University of Queensland, Brisbane, QLD, Australia.
Sci Rep. 2017 Aug 8;7(1):7521. doi: 10.1038/s41598-017-07749-w.
Rhizoctonia solani, the causal agent of rice sheath blight disease, causes significant losses worldwide as there are no cultivars providing absolute resistance to this fungal pathogen. We have used Host Delivered RNA Interference (HD-RNAi) technology to target two PATHOGENICITY MAP KINASE 1 (PMK1) homologues, RPMK1-1 and RPMK1-2, from R. solani using a hybrid RNAi construct. PMK1 homologues in other fungal pathogens are essential for the formation of appressorium, the fungal infection structures required for penetration of the plant cuticle, as well as invasive growth once inside the plant tissues and overall viability of the pathogen within the plant. Evaluation of transgenic rice lines revealed a significant decrease in fungal infection levels compared to non-transformed controls and the observed delay in disease symptoms was further confirmed through microscopic studies. Relative expression levels of the targeted genes, RPMK1-1 and RPMK1-2, were determined in R. solani infecting either transgenic or control lines with significantly lower levels observed in R. solani infecting transgenic lines carrying the HD-RNAi constructs. This is the first report demonstrating the effectiveness of HD-RNAi against sheath blight and offers new opportunities for durable control of the disease as it does not rely on resistance conferred by major resistance genes.
立枯丝核菌是引起水稻纹枯病的病原菌,由于目前尚无绝对抗该真菌病原体的品种,因此给全世界造成了巨大损失。我们使用宿主介导的 RNA 干扰(HD-RNAi)技术,针对立枯丝核菌中的两个致病性 MAPK1(PMK1)同源物 RPMK1-1 和 RPMK1-2,使用杂交 RNAi 构建体进行靶向操作。其他真菌病原体中的 PMK1 同源物对于形成附着胞至关重要,附着胞是真菌穿透植物角质层所需的侵染结构,以及在植物组织内的侵入性生长和病原体在植物体内的整体生存能力。对转基因水稻品系的评估表明,与非转化对照相比,真菌侵染水平显著降低,并且通过显微镜研究进一步证实了观察到的疾病症状延迟。在感染转基因或对照品系的立枯丝核菌中,靶向基因 RPMK1-1 和 RPMK1-2 的相对表达水平均有所降低,而在携带 HD-RNAi 构建体的转基因品系中感染的立枯丝核菌中观察到的水平明显更低。这是首次报道证明 HD-RNAi 对立枯丝核菌的有效性,并为该病的持久控制提供了新的机会,因为它不依赖于主要抗性基因赋予的抗性。