Abstract
Chitin is a linear polysaccharide composed of β (1→4)-linked N-acetylglucosamine (GlcNAc) residues. In fungi, chitin is an important component of the cell wall. Here, we provide a protocol to measure the chitin content of fungal cells using Magnaporthe oryzae as an example.
Keywords: Chitin, Cell wall, GlcNAc, Pathogenicity, Magnaporthe oryzae
Background
Chitin is an important component of the cell wall in fungal pathogens and is well known as a pathogen associated molecule pattern (PAMP). Determining the chitin content of a fungal species is important for studying fungal biology and host-pathogen interactions. The Morgan-Elson method based on colorimetric approach has been adapted in yeast to measure cellular chitin levels (Leloir and Cardini, 1953; Bulik et al., 2003; Baker et al., 2007). However, there is no specific protocol established for Magnaporthe oryzae, which is the causal agent of rice blast, the most important fungal disease in the world. Here, we describe a reliable and simple protocol which was modified from the Morgan-Elson method to test the chitin content of M. oryzae (Song et al., 2010).
Materials and Reagents
Equipment
Software
Procedure
Data analysis
Standard curves are prepared from stocks of 0.1 to 2.0 mM GlcNAc. Each result is presented at least three replicated measurements. The significance of differences between treatments is statistically evaluated using SDs and one-way analysis of variance (ANOVA) in SPSS 2.0 (Chicago, IL, USA). Data for two specific different treatments are compared statistically using ANOVA, followed by an F-test if the ANOVA result is significant at P < 0.05 or P < 0.01.
Notes
Recipes
Acknowledgments
This research was supported by the key program of Natural Science Foundation of China (Grant No. 31530063, ZZ), National Science Foundation for Distinguished Young Scholars of China (Grant No. 31325022 to ZZ), Natural Science Foundation of China (Grant No. 31271998, ZZ), and the especially appointed professorship (Jiangsu, China).
References
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