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<p>The present study focused on transcriptome profiling of in vitro regenerated <i>Stevia rebaudiana</i> genotypes to identify candidate genes involved in steviol glycosides under mannitol-induced stress. Two cultivars, CIM-Madhu and CIM-Mithi, were regenerated from nodal explants cultured on Murashige and Skoog (MS) medium with optimized concentrations of plant growth regulators. Maximum shoot regeneration (93.38%, 6.65 shoots/explant) was achieved with 1.0&#xa0;mg/L BAP and 0.5&#xa0;mg/L Kin, while rooting (96.67%) was highest with 1.0&#xa0;mg/L IAA. High-performance liquid chromatography (HPLC) analysis revealed a significant increase in stevioside content under 50&#xa0;g/L mannitol treatment, particularly in CIM-Mithi leaves (63.62&#xa0;µg/mL). Transcriptome sequencing generated over 35 million high-quality reads assembled into 61,988 transcripts, with CIM-Madhu showing more unigenes (2474) compared to CIM-Mithi (1851). Key biosynthetic genes (KO, UGT74G1, and UGT76G1) were identified, and their expression levels were validated through qPCR, which confirmed strong upregulation under mannitol stress, particularly for UGT74G1 (4.42-fold at 50&#xa0;g/L mannitol). These insights provide molecular targets for functional validation and open avenues for metabolic engineering and marker-assisted breeding to improve stevia’s commercial glycoside yield.</p>

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Transcriptomic Insights into Osmotic Stress-Mediated Regulation of Stevioside in In Vitro Stevia rebaudiana

  • 期刊论文

  • 发表日期：

    2025年11月26日

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  • Jay Gadhiya,
  • Rukam Singh Tomar,
  • Mahesh Mahatma,
  • P. J. Rathod,
  • Shraddha Bhatt,
  • Umesh Kandoliya,
  • H. P. Gajera,
  • Manoj V. Parakhia,
  • Ashishkumar G. Vala

摘要

The present study focused on transcriptome profiling of in vitro regenerated Stevia rebaudiana genotypes to identify candidate genes involved in steviol glycosides under mannitol-induced stress. Two cultivars, CIM-Madhu and CIM-Mithi, were regenerated from nodal explants cultured on Murashige and Skoog (MS) medium with optimized concentrations of plant growth regulators. Maximum shoot regeneration (93.38%, 6.65 shoots/explant) was achieved with 1.0 mg/L BAP and 0.5 mg/L Kin, while rooting (96.67%) was highest with 1.0 mg/L IAA. High-performance liquid chromatography (HPLC) analysis revealed a significant increase in stevioside content under 50 g/L mannitol treatment, particularly in CIM-Mithi leaves (63.62 µg/mL). Transcriptome sequencing generated over 35 million high-quality reads assembled into 61,988 transcripts, with CIM-Madhu showing more unigenes (2474) compared to CIM-Mithi (1851). Key biosynthetic genes (KO, UGT74G1, and UGT76G1) were identified, and their expression levels were validated through qPCR, which confirmed strong upregulation under mannitol stress, particularly for UGT74G1 (4.42-fold at 50 g/L mannitol). These insights provide molecular targets for functional validation and open avenues for metabolic engineering and marker-assisted breeding to improve stevia’s commercial glycoside yield.