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<p>Black rice (<i>Oryza sativa</i> L.) is known not only for its high anthocyanin levels but also for its potential carotenoid content. Despite Indonesia's rich diversity of black rice cultivars, information on their carotenoid profiles and gene expression remains limited. This present study investigated the β-carotene content in six local Indonesian black rice cultivars, the relative expression profiles of key carotenoid biosynthetic and regulatory genes, and the correlation between gene expression and β-carotene accumulation. Dehulled rice grains at the fully ripening stage (30 days after flowering, DAF) were used. β-carotene content was measured using high-performance liquid chromatography (HPLC), and relative gene expression was analyzed using quantitative real-time PCR (RT-qPCR). The target genes included <i>OsPSY</i>, <i>OsPDS</i>, <i>OsLCYe</i>, <i>OsLCYb</i>, <i>OsCYP97C</i>, <i>OsZEP</i>, <i>OsOr</i>, <i>OsbHLH</i>, and <i>OsWRKY</i>. Among the cultivars, Lampung Selatan and Cempo Ireng exhibited the highest β-carotene content (0.90 and 0.83 mg/kg, respectively), associated with elevated expression of upstream biosynthetic genes (<i>OsPSY, OsPDS, OsLCYe, OsLCYb, OsCYP97C</i>) and the regulatory gene <i>OsOr</i>. Correlation analysis revealed significant positive associations between β-carotene content and <i>OsPSY</i> (r = 0.573), <i>OsLCYb</i> (r = 0.572), <i>OsCYP97C</i> (r = 0.542), and <i>OsOr</i> (r = 0.583), while a negative correlation was found with <i>OsbHLH</i> (r = − 0.10). <i>OsZEP</i> expression was strongly correlated with regulatory genes <i>OsWRKY</i> (r = 0.730) and <i>OsbHLH</i> (r = 0.830). These findings highlight the complex transcriptional regulation of carotenoid metabolism in black rice and suggest molecular targets for biofortification through breeding or genome editing to enhance provitamin A content.</p>

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β-Carotene profiling and expression analysis of carotenoid biosynthetic genes in Indonesian black rice (Oryza sativa L.) cultivars

  • 期刊论文

  • 发表日期：

    2025年8月14日

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  • Ratih Restiani,
  • Tri Rini Nuringtyas,
  • Jose Gutierrez-Marcos,
  • Yekti Asih Purwestri

摘要

Black rice (Oryza sativa L.) is known not only for its high anthocyanin levels but also for its potential carotenoid content. Despite Indonesia's rich diversity of black rice cultivars, information on their carotenoid profiles and gene expression remains limited. This present study investigated the β-carotene content in six local Indonesian black rice cultivars, the relative expression profiles of key carotenoid biosynthetic and regulatory genes, and the correlation between gene expression and β-carotene accumulation. Dehulled rice grains at the fully ripening stage (30 days after flowering, DAF) were used. β-carotene content was measured using high-performance liquid chromatography (HPLC), and relative gene expression was analyzed using quantitative real-time PCR (RT-qPCR). The target genes included OsPSY, OsPDS, OsLCYe, OsLCYb, OsCYP97C, OsZEP, OsOr, OsbHLH, and OsWRKY. Among the cultivars, Lampung Selatan and Cempo Ireng exhibited the highest β-carotene content (0.90 and 0.83 mg/kg, respectively), associated with elevated expression of upstream biosynthetic genes (OsPSY, OsPDS, OsLCYe, OsLCYb, OsCYP97C) and the regulatory gene OsOr. Correlation analysis revealed significant positive associations between β-carotene content and OsPSY (r = 0.573), OsLCYb (r = 0.572), OsCYP97C (r = 0.542), and OsOr (r = 0.583), while a negative correlation was found with OsbHLH (r = − 0.10). OsZEP expression was strongly correlated with regulatory genes OsWRKY (r = 0.730) and OsbHLH (r = 0.830). These findings highlight the complex transcriptional regulation of carotenoid metabolism in black rice and suggest molecular targets for biofortification through breeding or genome editing to enhance provitamin A content.