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Asian Journal of Plant Sciences

Year: 2023 | Volume: 22 | Issue: 2 | Page No.: 309-315
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Authors

Theera Srisawat

Theera Srisawat

Yaowapa Sukpondma

Kanokrat Keawchai

Parinuch Chumkaew

Keywords

Research Article

Sodium Metabisulfite Suppresses Stoichiometry of Phenolic Compounds in Oil Palm Leaves for Flow Cytometric DNA Content Analysis

Theera Srisawat Theera Srisawat's LiveDNA, Yaowapa Sukpondma, Kanokrat Keawchai and Parinuch Chumkaew
Background and Objective: Nuclear lysis buffer containing sodium metabisulfite produces more efficient and accurate results from flow cytometric (FCM) analysis of a woody plant, but most studies have focused on only LB01 buffer. The present study aims to develop new nuclear lysis buffer used for flow cytometric analysis of oil palm. Materials and Methods: Phenolic contents in oil palms (three cultivars) were measured and evaluated for inhibition using Folin-Ciocalteu (FC) and High-Performance Liquid Chromatography (HPLC) analysis. Concentrations of sodium metabisulfite (0-20 mM) were modified with LB01-based buffer. The modified buffers were evaluated for effects on FCM analysis of oil palm. The repeatability of each experiment was performed for three replicates. Results: The phenolic compound found in both unopened leaves (UOL) and fully-opened leaves (FOL) of all seedling stages was tannic acid. Sodium metabisulfite (20 mM) was found to be an effective inhibitor of tannic acid by reducing the negative effects of tannic acid released from oil palm leaves, giving a high FL (276.30 channel units) and YF (0.35 nuclei s–1 mg–1) and the lowest CV (5.60%) for histograms. The LB01 led to satisfactory results in terms of DF and YF and the modified buffer containing 10-20 mM sodium metabisulfite was usually the most appropriate buffer for FCM analysis of oil palm leaves (2C DNA content = 3.7 pg). Conclusion: This study demonstrated the function of sodium metabisulfite, suggesting potential utility in oil palm DNA content analysis by FCM. Further experiments are needed to improve the buffer for oil palm tissue so that it can be applied directly to a wide variety of investigations.
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