Evaluasi Tiga Metode Preparasi RNA Total untuk Deteksi Turnip mosaic potyvirus dari Benih Brassica rappa dengan Reverse Transcription-Polymerase Chain Reaction

• Main Authors: Jati Adiputra, Sri Hendrastuti Hidayat, Tri Asmira Damayanti
• Format: Article
• Language: Indonesian
• Published: Perhimpunan Fitopatologi Indonesia 2013-04-01
• Series: Jurnal Fitopatologi Indonesia
• Subjects: Kata kunci: benih caisin, Brassica rappa, reverse transcription- polymerase chain reaction, RNA total, Turnip mosaic potyvirus
• Online Access: http://journal.ipb.ac.id/index.php/jfiti/article/view/6772

<p><em>Turnip mosaic virus (TuMV)</em> is an important seedborne virus that infects vegetable crops, especially Brassicae group, in Indonesia. Reliable detection method having high accuracy is required to detect the virus from seeds in order to avoid disease incidence as early as possible. A study was conducted to evaluate three total RNA extraction methods from <em>Brassica</em> seeds for TuMV detection using <em>reverse transcription- polymerase chain reaction</em>. The samples used are <em>Brassica rappa</em> seeds and seedling germinated for 3, 5, and 7 days. Total RNA extraction methods evaluated consisted of Wylie method, Randles method, and commercial kit as comparation. Total RNA extracted using Wylie and Randles methods has high concentration but low level of purity, on the other hand total RNA obtained using <span style="font-size: 10px;">commercial kit has low concentration but high level of purity. Detection of TuMV from seed was </span><span style="font-size: 10px;">successfully carried out using Wylie and Randles methods, TuMV from 3 - day old seedlings was detected using all three methodes, TuMV from 5-day old seedlings was only detected using Randles method, whereas TuMV from 7-day old seedlings was not detected using all three methodes. Total RNA extraction method using Wylie and Randles methodes is recommended for detection of TuMV from caisin seed using RT-PCR technique. </span></p><p><span style="font-size: 10px;">Key words: </span><em style="font-size: 10px;">Brassica rappa</em><span style="font-size: 10px;"> seed, </span><em style="font-size: 10px;">reverse transcription- polymerase chain reaction</em><span style="font-size: 10px;">, Total RNA, </span><em style="font-size: 10px;">Turnip mosaic potyvirus</em></p>