Table_1_Detection of Southern Rice Black-Streaked Dwarf Virus Using Western Blotting With P6.DOCX

The southern rice black-streaked dwarf virus (SRBSDV) is a severe threat to the yield and quality of rice products worldwide. Traditional detection methods for diagnosing SRBSDV infection show several false positives and thus provide inaccurate findings. However, Western blotting (WB) can precisely solve this problem. In this study, P6—a viral RNA-silencing suppressor—was expressed and purified in vitro. Two polyclonal P6 antibodies were obtained and quantified by enzyme-linked immunosorbent assay and WB. Subsequently, WB was performed using the P6 antibodies to identify SRBSDV antigens derived from the suspected rice samples collected from nine districts in Guizhou, China. The assay results showed that Libo, Pingtang, Huishui, Dushan, and Anshun districts had experienced an SRBSDV outbreak. The virus content in the sampled rice tissues was quantified by WB. Our results revealed that SRBSDV mainly accumulated in rice stems rather than rice leaves. Thus, the findings of our study show that the SRBSDV P6 antibody can be used in WB for detecting and monitoring SRBSDV infection in infected rice plants.

南方水稻黑条矮缩病毒（southern rice black-streaked dwarf virus, SRBSDV）对全球稻米产品的产量与品质均构成严重威胁。用于诊断SRBSDV感染的传统检测方法存在较多假阳性结果，因此检测结论不够准确。而蛋白质印迹法（Western blotting, WB）可精准解决这一问题。
本研究体外表达并纯化了P6蛋白——一种病毒RNA沉默抑制子。通过酶联免疫吸附试验与蛋白质印迹法，制备并定量得到两株P6多克隆抗体。随后，利用该P6抗体开展蛋白质印迹实验，对采自中国贵州省9个区县的疑似感染水稻样本中的SRBSDV抗原进行鉴定。
检测结果显示，荔波、平塘、惠水、独山及安顺等区县均暴发过SRBSDV疫情。通过蛋白质印迹法定量分析采样水稻组织中的病毒载量，结果表明SRBSDV主要积累于水稻茎秆中，而非水稻叶片。
本研究结果证实，SRBSDV P6抗体可应用于蛋白质印迹法，用于检测和监测感染水稻植株中的SRBSDV感染情况。