Construction of Fluorescently Labeled Strains of Nucleus and Mitochondria for Ustilago esculenta
TANG Jin-Tian*, YANG Fu-Rong*, ZHANG Lei-Lei, LI Yu-Kang, ZHANG Ya-Fen, FU Hui-Lan, XIA Wen-Qiang, CUI Hai-Feng, YE Zi-Hong**
College of Life Sciences/Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, HangZhou 310018, China
Abstract：Stem swollen of Zizania latifolia was associated with the successful infection of Ustilago esculenta, which depended on the dimorphic transition of U. esculenta. The dynamic changes of the nucleus and mitochondria during the process of dimorphic transition remain unknown. This study will focus on the coding gene for histone H1 and the coding sequence for the mitochondrial targeting signal (MTS) in the nucleus, respectively. After connecting them with the GFP and mCherry fluorescent protein genes, respectively, fluorescent labeling vectors for the nucleus and mitochondria was constructed. Wild-type strains UeT14 and UeT55 of U. esculenta as experimental materials to genetic transformation by Agrobacterium tumefaciens-mediated transformation (ATMT), and the recombinant strains that stably expressed red and green fluorescent proteins in nucleus or mitochondria were obtained: UeT55-H1-GFP, UeT14-H1-mCherry, UeT55-MTS-GFP and UeT14-MTS-mCherry. In UeT55-H1-GFP and UeT14-H1-mCherry strains, bright and round fluorescent spots could be seen in the center of the spores, which were co-located with the blue fluorescence formed by DAPI. In UeT55-MTS-GFP and UeT14-MTS-mCherry strains, small dot green or red fluorescence could be seen in the spores, and the green fluorescence was co-located with the red fluorescence formed by MitoTracker. In this study, the stable expression of fluorescent protein in nucleus or mitochondria was obtained, which provides an important reference and material for studying the molecular mechanism of growth, development, fusion and infection of U. esculenta.
汤近天, 杨芙容, 章蕾蕾, 李钰康, 张雅芬, 傅慧兰, 夏文强, 崔海峰, 叶子弘. 菰黑粉菌细胞核和线粒体的荧光标记菌株构建[J]. 农业生物技术学报, 2024, 32(5): 1188-1197.
TANG Jin-Tian, YANG Fu-Rong, ZHANG Lei-Lei, LI Yu-Kang, ZHANG Ya-Fen, FU Hui-Lan, XIA Wen-Qiang, CUI Hai-Feng, YE Zi-Hong. Construction of Fluorescently Labeled Strains of Nucleus and Mitochondria for Ustilago esculenta. 农业生物技术学报, 2024, 32(5): 1188-1197.
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