Rapid detection of the rice false smut fungus Ustilaginoidea virens by lateral flow strip-based recombinase polymerase amplification assay  

作　　者：Jiacheng Xi Sanlian Wan Yue Li Yuandi Xu Jing Yang Ting Zhang Jiajia Chen Zhengguang Zhang Danyu Shen Haifeng Zhang 

机构地区：[1]Sanya Institute of Nanjing Agricultural University/Key Laboratory of Integrated Management of Crop Diseases and Pests of Ministry of Education,Department of Plant Pathology,College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,China [2]Sanya Extension Service Center for Agricultural Technology,Sanya 572024,China [3]College of Landscape Architecture,Jiangsu Vocational College of Agriculture and Forestry,Zhenjiang 212400,China

出　　处：《Journal of Integrative Agriculture》2024年第11期3763-3773,共11页农业科学学报（英文版）

基　　金：supported by grants from the Jiangsu Agricultural Science and Technology Innovation Fund,China(JASTIF)(CX(21)3012)to Haifeng Zhang。

摘　　要：Rice false smut,caused by Ustilaginoidea virens,is a devastating disease that greatly reduces rice yield and quality.However,controlling rice false smut disease is challenging due to the unique infection mode of U.virens.Therefore,there is a need for early diagnosis and monitoring techniques to prevent the spread of this disease.Lateral flow strip-based recombinase polymerase amplification(LF-RPA)overcomes the limitations of current U.virens detection technologies,which are time-consuming,require delicate equipment,and have a high false-positive rate.In this study,we used a comparative genomics approach to identify Uv_3611,a specific gene of U.virens,as the target for the LF-RPA assay.The designed primers and probe efffectively detected the genomic DNA(gDNA)of U.virens and demonstrated no cross-reactivity with related pathogens.Under optimal conditions,the LF-RPA assay demonstrated a sensitivity of 10 pg of U.virens gDNA.Additionally,by incorporating a simplified PEG-NaOH method for plant DNA extraction,the LF-RPA assay enabled the detection of U.virens in rice spikelets within 30 min,without the need for specialized equipment.Furthermore,the LF-RPA assay successfully detected U.virens in naturally infected rice and seed samples in the field.Therefore,the LF-RPA assay is sensitive,efficient,and convenient,and could be developed as a kit for monitoring rice false smut disease in the field.

关 键 词：rice false smut Ustilaginoidea virens isothermal amplification disease monitoring field diagnosis 

分 类 号：S435.111.4[农业科学—农业昆虫与害虫防治]