Ray Soham, Singh Pankaj K, Gupta Deepak K, Mahato Ajay K, Sarkar Chiranjib, Rathour Rajeev, Singh Nagendra K, Sharma Tilak R
National Research Centre on Plant Biotechnology, Pusa Campus New Delhi, India.
Chaudhary Sarwan Kumar Himachal Pradesh Agricultural University Palampur, India.
Front Plant Sci. 2016 Aug 8;7:1140. doi: 10.3389/fpls.2016.01140. eCollection 2016.
Rice blast caused by Magnaporthe oryzae is one of the most important diseases of rice. Pi54, a rice gene that imparts resistance to M. oryzae isolates prevalent in India, was already cloned but its avirulent counterpart in the pathogen was not known. After decoding the whole genome of an avirulent isolate of M. oryzae, we predicted 11440 protein coding genes and then identified four candidate effector proteins which are exclusively expressed in the infectious structure, appresoria. In silico protein modeling followed by interaction analysis between Pi54 protein model and selected four candidate effector proteins models revealed that Mo-01947_9 protein model encoded by a gene located at chromosome 4 of M. oryzae, interacted best at the Leucine Rich Repeat domain of Pi54 protein model. Yeast-two-hybrid analysis showed that Mo-01947_9 protein physically interacts with Pi54 protein. Nicotiana benthamiana leaf infiltration assay confirmed induction of hypersensitive response in the presence of Pi54 gene in a heterologous system. Genetic complementation test also proved that Mo-01947_9 protein induces avirulence response in the pathogen in presence of Pi54 gene. Here, we report identification and cloning of a new fungal effector gene which interacts with blast resistance gene Pi54 in rice.
由稻瘟病菌引起的稻瘟病是水稻最重要的病害之一。Pi54是一个赋予水稻对在印度流行的稻瘟病菌株抗性的基因,已经被克隆,但该病原菌中与之对应的无毒基因尚不清楚。在对稻瘟病菌一个无毒分离株的全基因组进行解码后,我们预测了11440个蛋白质编码基因,然后鉴定出四个仅在侵染结构附着胞中表达的候选效应蛋白。通过计算机蛋白质建模,随后对Pi54蛋白模型与选定的四个候选效应蛋白模型进行相互作用分析,结果显示由位于稻瘟病菌4号染色体上的一个基因编码的Mo-01947_9蛋白模型,在Pi54蛋白模型的富含亮氨酸重复结构域处相互作用最佳。酵母双杂交分析表明Mo-01947_9蛋白与Pi54蛋白存在物理相互作用。烟草叶片浸润试验证实,在异源系统中存在Pi54基因时会诱导过敏反应。遗传互补试验也证明,在存在Pi54基因的情况下,Mo-01947_9蛋白在病原菌中诱导无毒反应。在此,我们报告了一个与水稻抗稻瘟病基因Pi54相互作用的新真菌效应基因的鉴定和克隆。