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<p><i>Picrorhiza kurroa</i> is an economically important high-altitude medicinal plant with huge therapeutic potential due to iridoid glycoside, i.e., picrosides. However, fragmented genomic information, the lack of genetic transformation protocols, and the unavailability of silencing methods limit the functional validation of key pathway genes <i>in planta</i> involved in picroside biosynthesis in <i>P. kurroa</i>. In this study, we established a method of virus-induced gene silencing (VIGS) in <i>P. kurroa</i> using a Tobacco rattle virus (TRV) derived vector and successfully silenced the phytoene desaturase (<i>Pk</i>PDS) gene. Virus-induced silencing of the <i>Pk</i>PDS gene results in a photobleaching phenotype in <i>P. kurroa.</i> The silencing of the <i>Pk</i>PDS gene was confirmed by the <i>Pk</i>PDS transcript, followed by an estimation of chlorophyll content in VIGS-treated leaves. RT-PCR analysis suggests a 65% reduction in transcript and a 60% reduction in total chlorophyll in VIGS leaves. Furthermore, we also observed a photobleaching phenotype in newly emerged leaves after infiltration, indicating that the silencing of <i>Pk</i>PDS spreads from one meristematic tissue to another. Thus, the current VIGS method might be employed to study the gene function involved in the biosynthesis of picrosides and other metabolites in <i>P. kurroa</i>. This aligns with United Nations Member States SDG-15.</p>

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Method development for virus-induced gene silencing in Picrorhiza kurroa, as an alternative tool for functional genomics

  • 期刊论文

  • 发表日期：

    2026年3月26日

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  • Monika Chouhan,
  • Vishal Saini,
  • Rajesh Kumar Singh,
  • Rajiv Kumar

摘要

Picrorhiza kurroa is an economically important high-altitude medicinal plant with huge therapeutic potential due to iridoid glycoside, i.e., picrosides. However, fragmented genomic information, the lack of genetic transformation protocols, and the unavailability of silencing methods limit the functional validation of key pathway genes in planta involved in picroside biosynthesis in P. kurroa. In this study, we established a method of virus-induced gene silencing (VIGS) in P. kurroa using a Tobacco rattle virus (TRV) derived vector and successfully silenced the phytoene desaturase (PkPDS) gene. Virus-induced silencing of the PkPDS gene results in a photobleaching phenotype in P. kurroa. The silencing of the PkPDS gene was confirmed by the PkPDS transcript, followed by an estimation of chlorophyll content in VIGS-treated leaves. RT-PCR analysis suggests a 65% reduction in transcript and a 60% reduction in total chlorophyll in VIGS leaves. Furthermore, we also observed a photobleaching phenotype in newly emerged leaves after infiltration, indicating that the silencing of PkPDS spreads from one meristematic tissue to another. Thus, the current VIGS method might be employed to study the gene function involved in the biosynthesis of picrosides and other metabolites in P. kurroa. This aligns with United Nations Member States SDG-15.