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<p>The synergistic fungicidal effect of radio frequency (RF) heating combined with natamycin against <i>A. niger</i> in rehydrated dried shiitake mushrooms has been demonstrated. However, the underlying molecular mechanism remains unclear. In this study, an <i>AnCAN33</i> deletion mutant (designated as the Δ<i>AnCAN33</i> mutant) was constructed via <i>Agrobacterium tumefaciens</i>-mediated homologous recombination to investigate its functional role in response to the combined treatment. Under the combined RF and natamycin treatment, the Δ<i>AnCAN33</i> mutant exhibited significantly higher inactivation efficiency than the wild-type strain, accompanied by aggravated membrane damage and increased leakage of intracellular contents. Deletion of <i>AnCAN33</i> markedly impaired polarized hyphal growth and altered spore surface ultrastructure, resulting in reduced mycelial biomass (0.09 ± 0.02&#xa0;g). In contrast, intracellular ATP accumulation increased to 0.388 ± 0.017&#xa0;µmol. RT-qPCR analysis revealed that transcriptional upregulation of homologous genes in the Δ<i>AnCAN33</i> mutant was insufficient to maintain normal physiological function under the combined treatment stress. In the absence of combined treatment, the Δ<i>AnCAN33</i> mutant exhibited significant alterations in hyphal growth and spore morphology, whereas sporulation and biomass remained largely unaffected compared to the wild-type. These results suggest that <i>AnCAN33</i> is associated with stress adaptation, membrane integrity, and cellular energy-related responses in <i>A. niger</i> under RF and natamycin stress. This study provides new insight into the molecular basis of the synergistic fungicidal effect of RF combined with natamycin.</p>

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Mechanistic Insight into the AnCAN33-mediated Response of Aspergillus niger to Radio Frequency Combined with Natamycin Treatment

  • 期刊论文

  • 发表日期：

    2026年7月24日

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  • Lihui Zhang,
  • Jialing Zhu,
  • Shaojin Wang,
  • Haile Ma,
  • Chunhua Dai,
  • Xiangyu Guan,
  • Yiting Guo

摘要

The synergistic fungicidal effect of radio frequency (RF) heating combined with natamycin against A. niger in rehydrated dried shiitake mushrooms has been demonstrated. However, the underlying molecular mechanism remains unclear. In this study, an AnCAN33 deletion mutant (designated as the ΔAnCAN33 mutant) was constructed via Agrobacterium tumefaciens-mediated homologous recombination to investigate its functional role in response to the combined treatment. Under the combined RF and natamycin treatment, the ΔAnCAN33 mutant exhibited significantly higher inactivation efficiency than the wild-type strain, accompanied by aggravated membrane damage and increased leakage of intracellular contents. Deletion of AnCAN33 markedly impaired polarized hyphal growth and altered spore surface ultrastructure, resulting in reduced mycelial biomass (0.09 ± 0.02 g). In contrast, intracellular ATP accumulation increased to 0.388 ± 0.017 µmol. RT-qPCR analysis revealed that transcriptional upregulation of homologous genes in the ΔAnCAN33 mutant was insufficient to maintain normal physiological function under the combined treatment stress. In the absence of combined treatment, the ΔAnCAN33 mutant exhibited significant alterations in hyphal growth and spore morphology, whereas sporulation and biomass remained largely unaffected compared to the wild-type. These results suggest that AnCAN33 is associated with stress adaptation, membrane integrity, and cellular energy-related responses in A. niger under RF and natamycin stress. This study provides new insight into the molecular basis of the synergistic fungicidal effect of RF combined with natamycin.