Molecular Identification of DNA Satellite Associated with Mungbean yellow mosaic India virus infecting Yardlong Bean in Yogyakarta

Gejala mosaik kuning dan keriting daun ditemukan pada pertanaman kacang panjang di Sleman, Yogyakarta. Begomovirus diketahui sebagai salah satu penyebab penyakit tersebut. Penelitian dilakukan untuk mengidentifikasi spesies Begomovirus dan DNA satelit yang berasosiasi dengan penyakit mosaik kuning kacang panjang. Ekstraksi DNA total dari tanaman bergejala dilanjutkan dengan amplifikasi fragmen DNA spesifik Begomovirus dan Betasatelit. Amplikon DNA berukuran ±1500 pb dan ±1300 pb berhasil diperoleh menggunakan berturut-turut primer universal Begomovirus dan primer spesifik Betasatelit. Analisis sekuen nukleotida mengonfirmasi identitas Begomovirus yang menginfeksi tanaman kacang panjang ialah Mungbean yellow mosaic India virus (MYMIV) dengan homologi 99% terhadap isolat MYMIV asal Indonesia. DNA satelit yang berasosiasi dengan MYMIV menunjukkan karakteristik Betasatelit, yaitu memiliki satellite common region (SCR) dengan struktur stem-loop dan sekuen TAATATTAC pada bagian loop, adenine rich region sebesar 54.96%, dan ORF (open reading frame) non-coding. Lebih lanjut, analisis rekombinasi menggunakan SimPlot mengindikasikan bahwa satelit non-coding MYMIV merupakan satelit rekombinan antara Betasatelit dan DNA-B Pepper yellow leaf curl Indonesia virus (PepYLCIV). Artikel ini merupakan laporan pertama asosiasi betasatelit DNA non-coding dengan MYMIV di Indonesia.Molecular Identification of Mungbean yellow mosaic India virus and Its Betasatellite Associated with Yellow Mosaic on Yardlong Bean in Sleman, Yogyakarta Yellow mosaic and leaf curl symptoms was observed in yard long baen cultivation in Sleman, Yogyakarta. Begomovirus is known as one of the causal agent of this disease.  Research was conducted to identify species of Begomovirus and  the associated DNA satellites from yellow mosaic disease of yard long bean. Total DNA was extracted from symptomatic leaves. DNA amplification was carried out through polymerase chain reaction using universal primer for Begomovirus and specific primer for Betasatellite.  DNA fragments of 1500 bp and 1300 bp was successfully amplified which indicated the presence of Begomovirus and DNA satellite in symptomatic leaves, respectively. Nucleotide sequence analysis showed that Begomovirus infecting yardlong bean has close relationship to Mungbean yellow mosaic India virus isolate from Indonesia with 99% homology. DNA satellite associated with MYMIV showed characteristic of Betasatellites, i.e. stem-loop structure in satellite conserve regions (SCR) TAATATTAC sequence in the loop region, adenine rich region (54.96%), and a non-coding ORF (open reading frame). SimPlot analysis suggests that the non-coding satellite associated with MYMIV was a recombinant satellite between betasatellite and DNA-B PepYLCIV (Pepper yellow leaf curl Indonesia virus). This study is the first report of the non-coding DNA satellite associated with MYMIV in Indonesia