Application of the Fluorescent Dye BODIPY in the Study of Lipid Dynamics of the Rice Blast Fungus Magnaporthe oryzae.
Application of the Fluorescent Dye BODIPY in the Study of Lipid Dynamics of the Rice Blast Fungus Magnaporthe oryzae.
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荧光染料BODIPY在稻瘟病菌脂质动力学研究中的应用
DOI：
10.3390/molecules23071594
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发表时间：
2018-06-30
期刊：
影响因子：
--
通讯作者：
Sun G
中科院分区：
文献类型：
--
作者：
Wang J;Guo X;Li L;Qiu H;Zhang Z;Wang Y;Sun G
稻瘟病是影响水稻产量的最严重病害之一，由稻瘟病菌（Magnaporthe oryzae）引起，稻瘟病菌是植物病原真菌研究的模式生物。米霉细胞中储存的脂质已被证明对于附着胞膨压的发展和病原体引起感染的能力至关重要。尼罗红染色是研究植物病原真菌脂质动态的常用方法。然而，该染料的缺点包括广谱、水溶性差、易猝灭。二吡咯亚甲基硼 (BODIPY) 是一种新型荧光染料，具有与尼罗红不同的发射波长以及许多理想的光谱和化学性质。在本研究中，我们使用 BODIPY 对米霉细胞中的脂质进行染色，以寻找尼罗红在植物病原真菌脂质动态研究中的可能替代品。我们的数据表明，通过简单和常规的程序，BODIPY 能够清楚地标记菌丝体和分生孢子细胞中的脂质。 BODIPY标记的脂质位置与尼罗红标记的脂质位置基本相同，但荧光标记更清晰，背景更低，特异性更高。使用 BODIPY 对发芽的米曲霉分生孢子进行染色，可以清楚地跟踪该过程中的脂质动态。我们还通过将 BODIPY 与红色荧光蛋白 mCherry 和其他荧光染料（例如 Calcoflor White 和 DAPI）相结合，在米霉的分生孢子、菌丝体和有性结构中实现了分生孢子的双重和多重荧光染色。这些结果表明BODIPY是一种理想的真菌脂质染色荧光染料，为植物病原真菌脂质动态和脂质代谢的研究提供了一种方法。
Rice blast is one of the most serious diseases affecting rice yield which is caused by Magnaporthe oryzae, a model organism for studies on plant pathogenic fungi. Lipids stored in M. oryzae cells have been shown to be crucial for the development of appressorium turgor and the ability of the pathogen to cause infection. Nile red staining is a common method to study lipid dynamics in phytopathogenic fungi. However, the disadvantages of this dye include its wide spectrum, poor water solubility, and susceptibility to quenching. Boron dipyrromethene (BODIPY) is a new type of fluorescent dye that has a different emission wavelength to that of Nile red as well as many desirable spectral and chemical properties. In this study, we used BODIPY to stain the lipids in M. oryzae cells to seek a possible substitute to Nile red in the study of lipid dynamics in plant pathogenic fungi. Our data showed that through simple and routine procedures, BODIPY was able to distinctly label lipids in the cells of mycelia and conidia. The positions of lipids labeled by BODIPY were essentially identical to those labeled by Nile red, but with more clear fluorescence labelling, lower background, and higher specificity. The use of BODIPY to stain germinating M. oryzae conidia allowed the lipid dynamics to be clearly tracked during this process. We also achieved double and multiple fluorescent staining conidia by combining BODIPY with the red fluorescent protein mCherry and other fluorescent dyes, such as Calcofluor white and DAPI, in conidia, mycelia, and sexual structures of M. oryzae. These results indicate that BODIPY is an ideal fluorescent dye for staining fungal lipids and provide a method for the study of the lipid dynamics and lipid metabolism in plant pathogenic fungi.
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影响因子：
2.9
作者：
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通讯作者：
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