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<p>It had been previously reported that trypsin was able to induce disease resistance and stress tolerance in <i>Hylocereus undatus</i> (<i>H. undatus</i>). However, how trypsin achieved the induction of resistance remained unclear. The results of this study showed that the regulated differential genes were mainly concentrated in the MYB transcription factor family after trypsin treatment. Transcriptomic analysis revealed that <i>HuMYBS3</i> was the core gene regulated by trypsin in the MYB family. Virus-induced gene silencing (VIGS) results showed that the total flavonoid content in the fruit decreased by 17% and fruit senescence significantly accelerated after HuMYBS3 was silenced. Transcriptomic data from silenced HuMYBS3 lines revealed significant changes in 2429 differentially expressed genes. Through Gene Set Enrichment Analysis (GSEA) combined with protein–protein interaction (PPI) network, <i>HuCYP84A1</i>, <i>HuCOMT1</i>, <i>HuFHT</i>, and <i>HuCHI</i> were further identified as likely key downstream target genes regulated by HuMYBS3. RT-qPCR verified the regulatory effect of HuMYBS3 on the four target genes. Further single-cell transcriptomic results showed spatial specificity in the expression of these four target genes of HuMYBS3. Except for <i>HuCYP84A1</i>, <i>HuCOMT1</i>, <i>HuFHT</i>, and <i>HuCHI</i> were highly expressed specifically in the exocarp. The results of the pseudotime analysis based on single-cell transcriptomic profiles revealed the molecular mechanisms by which HuMYBS3, activated by trypsin, regulated the two major downstream branches of lignin and flavonoids through <i>HuCOMT1</i> and <i>HuCHI</i>, respectively, in response to early stress. This study explored the mechanism by which trypsin induced resistance in <i>H. undatus</i> fruit and provided new theoretical support for exploring the preservation mechanism of <i>H. undatus</i>.</p>

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Revealing the regulation of HuMYBS3 on HuCOMT1 and HuCHI in response to biotic and abiotic stress through trypsin-induced resistance in pitaya using bulk and single-cell transcriptome profiles

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年3月3日

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  • Enyan Chen,
  • Jingyu Jia,
  • Jiaju Sun,
  • Xinxin Chen,
  • Fuxin Li,
  • Xin Li

摘要

It had been previously reported that trypsin was able to induce disease resistance and stress tolerance in Hylocereus undatus (H. undatus). However, how trypsin achieved the induction of resistance remained unclear. The results of this study showed that the regulated differential genes were mainly concentrated in the MYB transcription factor family after trypsin treatment. Transcriptomic analysis revealed that HuMYBS3 was the core gene regulated by trypsin in the MYB family. Virus-induced gene silencing (VIGS) results showed that the total flavonoid content in the fruit decreased by 17% and fruit senescence significantly accelerated after HuMYBS3 was silenced. Transcriptomic data from silenced HuMYBS3 lines revealed significant changes in 2429 differentially expressed genes. Through Gene Set Enrichment Analysis (GSEA) combined with protein–protein interaction (PPI) network, HuCYP84A1, HuCOMT1, HuFHT, and HuCHI were further identified as likely key downstream target genes regulated by HuMYBS3. RT-qPCR verified the regulatory effect of HuMYBS3 on the four target genes. Further single-cell transcriptomic results showed spatial specificity in the expression of these four target genes of HuMYBS3. Except for HuCYP84A1, HuCOMT1, HuFHT, and HuCHI were highly expressed specifically in the exocarp. The results of the pseudotime analysis based on single-cell transcriptomic profiles revealed the molecular mechanisms by which HuMYBS3, activated by trypsin, regulated the two major downstream branches of lignin and flavonoids through HuCOMT1 and HuCHI, respectively, in response to early stress. This study explored the mechanism by which trypsin induced resistance in H. undatus fruit and provided new theoretical support for exploring the preservation mechanism of H. undatus.