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<p>Famous Iranian rice cultivars are known for their unique tastes and aroma. This study is the first to compare volatile terpenes among these traditional rice types, assessing terpene concentrations and antioxidant activity. Three famous rice cultivars “Champa” (CH), “Hashemi” (HA), and “Shamim” (SH) were selected for this experiment. We qualified and quantified terpene concentrations, assessed antioxidant activity, and performed RNA-seq to identify differentially expressed genes (DEGs) related to terpenoid and antioxidant pathways. Thin-layer chromatography (TLC) analysis showed varying Rf values for total terpenes, while catalase (CAT) assays indicated significantly higher spectrophotometric values for HA. Glutathione S-transferase (GST), peroxidase (PRX), and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assays showed significantly higher spectrophotometric values for SH (<i>p</i> &lt; 0.05). RNA-seq analysis of terpenoid and antioxidant enzyme-coding genes was performed in three comparisons: CH vs. HA, CH vs. SH, and SH vs. HA, with CH as the control. Differentially expressed genes (DEGs) from the PRX, GST, and terpene synthase (TPS) gene families were identified. The hierarchical clustering of heatmap results clearly distinguished HA and SH regarding TPS and PRX. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses revealed associations with phenylpropanoid biosynthesis, glutathione metabolism, and antioxidants. Terpenoid biosynthesis pathways were enriched in the CH vs. HA and SH vs. HA comparisons, while the CH vs. SH variation was enriched in glutathione metabolism. Transcriptome data revealed distinct expression patterns, with HA significantly differing from CH and SH. This research represents RNA sequencing analysis of Iranian rice cultivars, offering insights into enhancing aromatic quality, volatile terpenoid accumulation, and pest resistance. It establishes a genomic foundation for investigating the molecular mechanisms behind the medicinal properties of these rice cultivars. Moreover, it emphasizes that differential expression of terpenoid biosynthesis genes significantly influences the aromatic profiles of rice cultivars. These results may benefit future experiments on the antioxidant activity and flavor enhancement of rice cultivars.</p>

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High-throughput RNA sequencing and thin-layer chromatography provide insights into the molecular basis of distinctive scents and antioxidant capacity differences in Iranian rice cultivars

  • 期刊论文

  • 发表日期：

    2025年5月29日

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  • Seyedeh-Mahsa Safieddinardebili,
  • Iraj Mehregan,
  • Reza Azizinezhad,
  • Sara Saadatmand,
  • Rahim Ahmadvand

摘要

Famous Iranian rice cultivars are known for their unique tastes and aroma. This study is the first to compare volatile terpenes among these traditional rice types, assessing terpene concentrations and antioxidant activity. Three famous rice cultivars “Champa” (CH), “Hashemi” (HA), and “Shamim” (SH) were selected for this experiment. We qualified and quantified terpene concentrations, assessed antioxidant activity, and performed RNA-seq to identify differentially expressed genes (DEGs) related to terpenoid and antioxidant pathways. Thin-layer chromatography (TLC) analysis showed varying Rf values for total terpenes, while catalase (CAT) assays indicated significantly higher spectrophotometric values for HA. Glutathione S-transferase (GST), peroxidase (PRX), and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assays showed significantly higher spectrophotometric values for SH (p < 0.05). RNA-seq analysis of terpenoid and antioxidant enzyme-coding genes was performed in three comparisons: CH vs. HA, CH vs. SH, and SH vs. HA, with CH as the control. Differentially expressed genes (DEGs) from the PRX, GST, and terpene synthase (TPS) gene families were identified. The hierarchical clustering of heatmap results clearly distinguished HA and SH regarding TPS and PRX. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analyses revealed associations with phenylpropanoid biosynthesis, glutathione metabolism, and antioxidants. Terpenoid biosynthesis pathways were enriched in the CH vs. HA and SH vs. HA comparisons, while the CH vs. SH variation was enriched in glutathione metabolism. Transcriptome data revealed distinct expression patterns, with HA significantly differing from CH and SH. This research represents RNA sequencing analysis of Iranian rice cultivars, offering insights into enhancing aromatic quality, volatile terpenoid accumulation, and pest resistance. It establishes a genomic foundation for investigating the molecular mechanisms behind the medicinal properties of these rice cultivars. Moreover, it emphasizes that differential expression of terpenoid biosynthesis genes significantly influences the aromatic profiles of rice cultivars. These results may benefit future experiments on the antioxidant activity and flavor enhancement of rice cultivars.