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<p><i>Catharanthus roseus</i> is known for producing anticancer monoterpenoid indole alkaloids (MIAs) vincristine and vinblastine. These alkaloids are synthesized in a complex pathway with several catalyzing steps. <i>Trans</i>-isoprenyl diphosphate synthase (TIDS) is an important family of enzymes that catalyze the initial biosynthesis steps of MIA production. However, comprehensive characterization of the TIDS gene family has not yet been done in <i>C. roseus</i>. A combination of computational and laboratory techniques was used in the current study to ascertain the role of the TIDS family under cold, hormonal, wounding, and yeast extract stress conditions. In silico analyses of the identified genes included motif and gene structure analysis, prediction of cis-acting elements, phylogenetic, evolutionary, and expression analysis. The homology modeling approach identified 4 homologs each of <i>geranyl diphosphate synthase</i> (<i>GPPS</i>) and <i>geranylgeranyl diphosphate synthase</i> (<i>GGPPS</i>), resulting in a total of 8 <i>TIDS</i> genes in the genome of <i>C. roseus</i>. Furthermore, the <i>Cr</i>TIDS proteins were clustered into four phylogenetic groups. Expression studies using qRT-PCR revealed the highest expression of <i>CrGPPS-01</i> in response to 1&#xa0;g/L yeast extract (119-fold), followed by cold stress for 72&#xa0;h (112-fold). The phenomenal upregulation of the <i>CrGPPS-01</i> points to the involvement of this homolog in regulating stress conditions culminating in MIA production. The results of the present study offer important insights into the <i>CrTIDS</i> gene family and a step forward in shaping the regulatory checkpoints for an improved biosynthesis of desired MIAs solely produced by <i>C. roseus</i>.</p>

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Expression Profiling of Genome-Wide Identified and Characterized Trans-Isoprenyl Diphosphate Synthase Family in Catharanthus roseus (L.) G. Don Under Stress Conditions

  • 期刊论文

  • 发表日期：

    2025年5月21日

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  • Muhammad Huzafa,
  • Amjad Hassan,
  • Batool Bashir,
  • Azhar Mehmood,
  • Jawad Ahmed

摘要

Catharanthus roseus is known for producing anticancer monoterpenoid indole alkaloids (MIAs) vincristine and vinblastine. These alkaloids are synthesized in a complex pathway with several catalyzing steps. Trans-isoprenyl diphosphate synthase (TIDS) is an important family of enzymes that catalyze the initial biosynthesis steps of MIA production. However, comprehensive characterization of the TIDS gene family has not yet been done in C. roseus. A combination of computational and laboratory techniques was used in the current study to ascertain the role of the TIDS family under cold, hormonal, wounding, and yeast extract stress conditions. In silico analyses of the identified genes included motif and gene structure analysis, prediction of cis-acting elements, phylogenetic, evolutionary, and expression analysis. The homology modeling approach identified 4 homologs each of geranyl diphosphate synthase (GPPS) and geranylgeranyl diphosphate synthase (GGPPS), resulting in a total of 8 TIDS genes in the genome of C. roseus. Furthermore, the CrTIDS proteins were clustered into four phylogenetic groups. Expression studies using qRT-PCR revealed the highest expression of CrGPPS-01 in response to 1 g/L yeast extract (119-fold), followed by cold stress for 72 h (112-fold). The phenomenal upregulation of the CrGPPS-01 points to the involvement of this homolog in regulating stress conditions culminating in MIA production. The results of the present study offer important insights into the CrTIDS gene family and a step forward in shaping the regulatory checkpoints for an improved biosynthesis of desired MIAs solely produced by C. roseus.