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<p>Subcellular localization of focal proteins is essential for elucidating protein function. Using <i>Agrobacterium</i>-mediated transformation, we developed stable lines of transgenic rice (<i>Oryza sativa</i>) that express fluorescent protein-tagged organelle markers designed to target the endoplasmic reticulum, Golgi apparatus, filamentous actin, autophagosome, plasma membrane, and nucleus. These markers can generate valuable data for investigating protein localization and organelle dynamics. To explore autophagy under nutrient stress, we used a mCherry:OsATG8a marker line to examine autophagosome formation under phosphate-deficient conditions. We found a significant increase in autophagosomes during phosphate starvation, which was further confirmed by co-localization with monodansylcadaverine (MDC), a known marker of the autophagosome. Overall, our findings demonstrate that phosphate deficiency enhances autophagy in rice and establish that fluorescent protein-tagged organelle markers are powerful tools for studying subcellular dynamics under both physiological and stress conditions.</p>

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Stable Expression of Organelle-Targeted Fluorescent Markers for Investigating Autophagy and Subcellular Dynamics in Rice

  • 期刊论文

  • 发表日期：

    2025年9月10日

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  • Yun-Shil Gho,
  • Seo-Youn Park,
  • Heebak Choi,
  • Minseo Kang,
  • Woojung Yang,
  • Su Kyoung Lee,
  • Sunok Moon,
  • Sun-Hwa Ha,
  • Ki-Hong Jung

摘要

Subcellular localization of focal proteins is essential for elucidating protein function. Using Agrobacterium-mediated transformation, we developed stable lines of transgenic rice (Oryza sativa) that express fluorescent protein-tagged organelle markers designed to target the endoplasmic reticulum, Golgi apparatus, filamentous actin, autophagosome, plasma membrane, and nucleus. These markers can generate valuable data for investigating protein localization and organelle dynamics. To explore autophagy under nutrient stress, we used a mCherry:OsATG8a marker line to examine autophagosome formation under phosphate-deficient conditions. We found a significant increase in autophagosomes during phosphate starvation, which was further confirmed by co-localization with monodansylcadaverine (MDC), a known marker of the autophagosome. Overall, our findings demonstrate that phosphate deficiency enhances autophagy in rice and establish that fluorescent protein-tagged organelle markers are powerful tools for studying subcellular dynamics under both physiological and stress conditions.