Use of chitin:DNA ratio to assess growth form in fungal cells.
Use of chitin:DNA ratio to assess growth form in fungal cells.
复制标题
DOI：
10.1186/s12915-024-01815-2
复制
发表时间：
2024-01-17
期刊：
影响因子：
5.4
通讯作者：
Fones, Helen N.
中科院分区：
文献类型：
--
作者：
Kovacs-Simon, Andrea;Fones, Helen N.
二形性是某些真菌的一个重要特征，包括重要的植物和人类病原体，它能够在类酵母和菌丝生长形式之间切换。向菌丝生长的转变通常与毒力、致病性、生物膜形成和抗逆性有关。因此，准确和有效地测量真菌生长形态的能力是研究这些真菌，特别是发现潜在的药物靶点的关键。到目前为止，真菌生长形式是通过显微镜进行评估的，这一过程既是劳动密集型的，又是昂贵的。在这里，我们结合量化真菌细胞壁中的甲壳素和细胞核中的DNA来产生一种方法，该方法允许通过计算真菌或受感染宿主组织样本中存在的细胞壁数量和细胞核数量之间的比率来估计真菌细胞的形状。以小麦病原菌ZymosepVictoria tritici为例，证实了几丁质与DNA的比值与真菌细胞的平均极性指数(长/宽)之间存在密切的线性关系。我们展示了该方法在估计受感染的小麦叶片中生长形态的实用性，并使用这一比率区分了两个不同的小麦纹枯病菌菌株的萌发时间。我们还表明，该方法对厚壁孢子的发生是稳健的，厚壁孢子显示出几丁质：DNA的比率，既不同于酵母样的芽生孢子，也不同于菌丝。甲壳素：DNA比率提供了一种简单的方法来确定大量组织样本中的真菌生长形式，减少了需要特定染色或GFP标签来可视化宿主组织中的真菌的劳动密集型显微镜研究的需要。它适用于各种实验条件下的二相性真菌。网上版载有补充材料，可在10.1186/s12915-024-01815-2查阅。
Dimorphism, the ability to switch between a ‘yeast-like’ and a hyphal growth form, is an important feature of certain fungi, including important plant and human pathogens. The switch to hyphal growth is often associated with virulence, pathogenicity, biofilm formation and stress resistance. Thus, the ability to accurately and efficiently measure fungal growth form is key to research into these fungi, especially for discovery of potential drug targets. To date, fungal growth form has been assessed microscopically, a process that is both labour intensive and costly. Here, we unite quantification of the chitin in fungal cell walls and the DNA in nuclei to produce a methodology that allows fungal cell shape to be estimated by calculation of the ratio between cell wall quantity and number of nuclei present in a sample of fungus or infected host tissue. Using the wheat pathogen Zymoseptoria tritici as a test case, with confirmation in the distantly related Fusarium oxysporum, we demonstrate a close, linear relationship between the chitin:DNA ratio and the average polarity index (length/width) of fungal cells. We show the utility of the method for estimating growth form in infected wheat leaves, differentiating between the timing of germination in two different Z. tritici isolates using this ratio. We also show that the method is robust to the occurrence of thick-walled chlamydospores, which show a chitin:DNA ratio that is distinct from either ‘yeast-like’ blastospores or hyphae. The chitin:DNA ratio provides a simple methodology for determining fungal growth form in bulk tissue samples, reducing the need for labour-intensive microscopic studies requiring specific staining or GFP-tags to visualise the fungus within host tissues. It is applicable to a range of dimorphic fungi under various experimental conditions. The online version contains supplementary material available at 10.1186/s12915-024-01815-2.
登录
查看更多内容
影响因子：
4.6
作者：
Fones HN
通讯作者：
Fones HN
影响因子：
--
作者：
Duncan, KE;Howard, RJ
通讯作者：
Howard, RJ
影响因子：
3
作者：
Francisco，Carolina Sardinha;McDonald，Bruce A.;Palma-Guerrero，Javier
通讯作者：
Palma-Guerrero，Javier
影响因子：
2.1
作者：
Guo, Jian-Rong;Schnieder, Frank;Verreet, Joseph-Alexander
通讯作者：
Verreet, Joseph-Alexander
影响因子：
3
作者：
Fones, H. N.;Soanes, D.;Gurr, S. J.
通讯作者：
Gurr, S. J.