All-in-one comprehensive extraction of metabolites, proteins and ribonucleic acid for the rapid analysis of oil palm systems biology

Nurazah Zain; Abrizah Othman; Nurul Liyana Rozali; Umi Salamah Ramli; Syahanim Shahwan

All-in-one comprehensive extraction of metabolites, proteins and ribonucleic acid for the rapid analysis of oil palm systems biology

2022

Nurazah Zain | Abrizah Othman | Nurul Liyana Rozali | Umi Salamah Ramli | Syahanim Shahwan

Oil palm (Elaeis guineensis Jacq.) systems biology offers a comprehensive view of the plant system by employing a holistic multi-omics approach encompassing the molecular data at various hierarchical levels. Sample limitation and the importance of integrating all molecular data with minimal variation, led to the development of sequential extraction of biomolecules fractions from a single undivided biological sample. Oil palm root was subjected to solvent extraction with methanol-chloroform-water to recover metabolites of diverse polarity. The resultant pellet was subjected to buffer and solvent partitioning to obtain RNA and proteins. RNA extracted from the oil palm root showed a recovery of 180.25 ng mg–¹, with an A260:A280 ratio ranging between 1.9-2.0 and RNA integrity number (RIN) value of 6.7. Co-extracted proteins resulted in a recovery of 29.28 µg per 100 mg fresh weight (FW) tissue and revealed a total of 1852 identified proteins in the oil palm root. Polar metabolites revealed approximately 40 metabolite peaks, and non-polar metabolites with two major fatty acid groups i.e., saturated and unsaturated fatty acids at 55.4% and 38.6%, respectively. This protocol demonstrated an advancement of extraction protocols for oil palm root biomolecules, which will consecutively expedite the establishment of various multi-omics platforms.

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