State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China.
College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China.
Molecules. 2018 Jun 30;23(7):1594. doi: 10.3390/molecules23071594.
Rice blast is one of the most serious diseases affecting rice yield which is caused by , a model organism for studies on plant pathogenic fungi. Lipids stored in 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 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 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 . 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.
稻瘟病是影响水稻产量的最严重疾病之一，由 引起，它是研究植物病原真菌的模式生物。已表明，储存于 细胞中的脂质对于形成附着胞膨压和病原菌致病能力至关重要。尼罗红染色是研究植物病原真菌中脂质动态的常用方法。然而，这种染料的缺点包括其广谱性、水溶性差和易淬灭。硼二吡咯甲川（BODIPY）是一种新型荧光染料，与尼罗红相比，其发射波长不同，并且具有许多理想的光谱和化学性质。在这项研究中，我们使用 BODIPY 对 细胞中的脂质进行染色，以寻找一种可能替代尼罗红的方法，用于研究植物病原真菌中的脂质动态。我们的数据表明，通过简单常规的程序，BODIPY 能够清晰地标记菌丝体和孢子细胞中的脂质。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 对萌发的 孢子进行染色，可以在这个过程中清楚地跟踪脂质的动态变化。我们还通过将 BODIPY 与红色荧光蛋白 mCherry 以及其他荧光染料（如 Calcofluor white 和 DAPI）结合，在孢子、菌丝体和 有性结构中实现了双荧光和多荧光染色的孢子。这些结果表明，BODIPY 是一种理想的真菌脂质荧光染料，为研究植物病原真菌中的脂质动态和脂质代谢提供了一种方法。
