Two dimensional electrophoresis application for Ganoderma study in oil palm
2007
Syahanim Shahwan, Malaysian Palm Oil Board (MPOB), 6, Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor (Malaysia)
Crop plants have become a major focus world wide today due to their ability to retain the economical status of the country. However, the major problem facing the crop industry is the infection by various pathogens such as fungus. Root and stem rots caused by Ganoderma have been known to cause extensive losses of many crops. And now, the fungus threats the oil palm industry. Study was conducted to develop method for extracting total proteins in the oil palm root. Total protein was extracted using various methods. As a preliminary study, protein was extracted based on three different methods: Giavalisco et al., 2003 (M1), Modified method by Giavaliso et al., 2003 (M2) and Sheffield et al., 2006 (M3). The M1 protocol showed good quality spots with reduced background for primary roots. However, this method was applied to the secondary roots and heavy vertical streaking was observed at the acidic part of the strip. M2 was conducted to reduce this pr oblem, however, the vertical streaking was still seen at the acidic segment. Therefore, M3 was conducted and vertical streaking problem successfully reduced. Extracted protein was resolved using broad range (pH 3-10) IPG strips in the first dimension. Then, second dimension was conducted using 10% SDS-P AG E gels. Gels were stained with Silver Staining Kit (Bio-Rad, Hercules, CA) and the spots will be analyzed using PO-Quest Software (Bio-Rad, Hercules, CA).
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