Metode DAS-ELISA dan RT-PCR untuk Deteksi Potyvirus yang Menginfeksi Bawang Merah
DAS-ELISA and RT-PCR Methods for Detection of Potyviruses Infecting Shallot
DOI:
https://doi.org/10.14692/jfi.20.5.255-262Keywords:
cross-reaction, Onion yellow dwarf virus, RT-PCR, sequence analysis, Shallot yellow stripe virusAbstract
DAS-ELISA and RT-PCR Methods for Detection of Potyviruses Infecting Shallot
Viral infection of shallots in Indonesia may reduce yields, mainly because it is bulb-borne. Virus detection to monitor the incidence of disease in the field or examine healthy seeds requires an accurate and reliable method. In this study, the detection of Potyvirus members, i.e., Onion yellow dwarf virus (OYDV) and Shallot yellow stripe virus (SYSV), was conducted using enzyme-linked immunosorbent assay (DAS-ELISA) and reverse transcription polymerase chain reaction (RT-PCR) from shallot leaf samples. Leaf samples were collected from several shallot growing areas in Brebes and Cirebon. Infections of OYDV and SYSV were detected using the DAS-ELISA method, then field samples that gave positive reaction were selected for further detection using the RT-PCR method, followed by nucleotide sequencing. Based on the DAS-ELISA method, OYDV and SYSV infections were detected from field samples as single and mixed infections. Potyvirus infection was confirmed by the RT-PCR method from field samples that gave a positive reaction in ELISA. Nucleotide sequence analysis showed that OYDV infection was detected in both samples that reacted positively with OYDV and SYSV antiserum. These results indicated the cross-reaction between OYDV and SYSV antisera.
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