Functional validation of pathogenicity genes in rice sheath blight pathogen Rhizoctonia solani by a novel host-induced gene silencing system,Molecular Plant Pathology

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Functional validation of pathogenicity genes in rice sheath blight pathogen Rhizoctonia solani by a novel host-induced gene silencing system
Molecular Plant Pathology ( IF 4.8 ) Pub Date : 2021-08-27 , DOI: 10.1111/mpp.13130
Mei Zhao ₁ , Chenjiaozi Wang ₁ , Jun Wan ₁ , Zanfeng Li ₁ , Dilin Liu ₂ , Naoki Yamamoto ₃ , Erxun Zhou ₁ , Canwei Shu ₁

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Rice sheath blight, caused by the soilborne fungus Rhizoctonia solani, causes severe yield losses worldwide. Elucidation of the pathogenic mechanism of R. solani is highly desired. However, the lack of a stable genetic transformation system has made it challenging to examine genes' functions in this fungus. Here, we present functional validation of pathogenicity genes in the rice sheath blight pathogen R. solani by a newly established tobacco rattle virus (TRV)–host-induced gene silencing (HIGS) system using the virulent R. solani AG-1 IA strain GD-118. RNA interference constructs of 33 candidate pathogenicity genes were infiltrated into Nicotiana benthamiana leaves with the TRV-HIGS system. Of these constructs, 29 resulted in a significant reduction in necrosis caused by GD-118 infection. For further validation of one of the positive genes, trehalose-6-phosphate phosphatase (Rstps2), stable rice transformants harbouring the double-stranded RNA (dsRNA) construct for Rstps2 were created. The transformants exhibited reduced gene expression of Rstps2, virulence, and trehalose accumulation in GD-118. We showed that the dsRNA for Rstps2 was taken up by GD-118 mycelia and sclerotial differentiation of GD-118 was inhibited. These findings offer gene identification opportunities for the rice sheath blight pathogen and a theoretical basis for controlling this disease by spray-induced gene silencing.

中文翻译：

一种新型宿主诱导基因沉默系统对水稻纹枯病病原体立枯丝核菌致病基因的功能验证

由土壤传播的真菌立枯丝核菌引起的水稻纹枯病在全球范围内造成严重的产量损失。非常需要阐明R. solani的致病机制。然而，由于缺乏稳定的遗传转化系统，因此很难检测这种真菌中的基因功能。在这里，我们通过新建立的烟草脆裂病毒 (TRV)-宿主诱导的基因沉默 (HIGS) 系统使用毒性R.对水稻纹枯病病原体R. solani的致病基因进行了功能验证。茄尼AG-1 IA 菌株 GD-118。33个候选致病基因的RNA干扰构建体被渗透到本氏烟草中离开 TRV-HIGS 系统。在这些构建体中，29 个导致由 GD-118 感染引起的坏死显着减少。为了进一步验证阳性基因之一，海藻糖-6-磷酸磷酸酶( Rstps2 )，产生了含有Rstps2双链 RNA (dsRNA) 构建体的稳定水稻转化体。转化体在GD-118中表现出Rstps2基因表达、毒力和海藻糖积累降低。我们展示了Rstps2的 dsRNA被GD-118菌丝体吸收，GD-118的菌核分化受到抑制。这些发现为水稻纹枯病病原体的基因鉴定提供了机会，并为通过喷雾诱导的基因沉默控制该病害提供了理论基础。

更新日期：2021-08-27

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