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<p>The E3 ubiquitin ligase HOS1 is a key negative regulator of the ICE1–CBF/DREB cold signaling pathway in <i>Arabidopsis thaliana</i> and participates in various stress and developmental processes. While its role in cold tolerance is well established, its influence on secondary metabolism and multi-stress responses in plant cells remains unknown. Here, we employed CRISPR/Cas9 genome editing to generate <i>hos1</i> knockout <i>Arabidopsis</i> callus lines and investigated their growth dynamics, abiotic stress tolerance, indole glucosinolate (IGS) metabolism, and transcriptional reprogramming. Mutant lines exhibited enhanced biomass accumulation, increased cold and osmotic stress tolerance, and unexpectedly improved heat tolerance. qRT-PCR revealed activation of ICE1-CBF/DREB and ABA signaling modules, along with strong induction of heat shock transcription factors. HPLC-MS showed up to a 60-fold increase in 1-methoxyindol-3-ylmethyl glucosinolate and 4-hydroxyindol-3-ylmethyl, accompanied by reduced levels of their precursor, indol-3-ylmethyl glucosinolate. These changes were linked to elevated expression of MYB/MYC regulators and corresponding biosynthetic genes. Cold stress amplified IGS accumulation, while heat stress fully suppressed it. The results indicate that HOS1 disruption rewires stress signaling and metabolism in a stress-specific manner, suggesting a biotechnological route to improve resilience and metabolite yields in plant cells.</p>

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HOS1 knockout enhances glucosinolate biosynthesis and activates multi-stress signaling in Arabidopsis callus

  • 期刊论文

  • 发表日期：

    2026年3月28日

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  • Y. A. Yugay,
  • O. V. Grishchenko,
  • V. P. Grigorchuk,
  • A. V. Fialko,
  • G. N. Veremeichik,
  • T. V. Rusapetova,
  • Y. N. Shkryl,
  • V. P. Bulgakov

摘要

The E3 ubiquitin ligase HOS1 is a key negative regulator of the ICE1–CBF/DREB cold signaling pathway in Arabidopsis thaliana and participates in various stress and developmental processes. While its role in cold tolerance is well established, its influence on secondary metabolism and multi-stress responses in plant cells remains unknown. Here, we employed CRISPR/Cas9 genome editing to generate hos1 knockout Arabidopsis callus lines and investigated their growth dynamics, abiotic stress tolerance, indole glucosinolate (IGS) metabolism, and transcriptional reprogramming. Mutant lines exhibited enhanced biomass accumulation, increased cold and osmotic stress tolerance, and unexpectedly improved heat tolerance. qRT-PCR revealed activation of ICE1-CBF/DREB and ABA signaling modules, along with strong induction of heat shock transcription factors. HPLC-MS showed up to a 60-fold increase in 1-methoxyindol-3-ylmethyl glucosinolate and 4-hydroxyindol-3-ylmethyl, accompanied by reduced levels of their precursor, indol-3-ylmethyl glucosinolate. These changes were linked to elevated expression of MYB/MYC regulators and corresponding biosynthetic genes. Cold stress amplified IGS accumulation, while heat stress fully suppressed it. The results indicate that HOS1 disruption rewires stress signaling and metabolism in a stress-specific manner, suggesting a biotechnological route to improve resilience and metabolite yields in plant cells.