Development of SYBR Green I‐Based One‐Step Real Time RT‐PCR Assay for Quantifying Southern rice black‐streaked dwarf virus in Rice

Abstract Southern rice black‐streaked dwarf virus ( SRBSDV ) causes southern rice black‐streaked dwarf and maize rough dwarf diseases, which lead to severe yield losses of crops in Southeast Asia. We report here a SYBR G reen I ‐based O ne‐ S tep R eal T ime RT ‐ PCR assay for quantifying SRBSDV in rice rapidly and accurately. Primers used for assay were designed from the conserved sequence in S 9 RNA among SRBSDV isolates. The RNA standards targeting the S 9 region were obtained by transcription in vitro for generation of a standard curve. The assay developed in this study was found to be 100 times more sensitive than the conventional RT ‐ PCR for SRBSDV detection. The primers were very specific for SRBSDV . This study clearly demonstrated the potential usefulness of developed assay for detection and quantitation of SRBSDV in rice samples.