The Plant Pathology Journal

The Korean Society of Plant Pathology (한국식물병리학회)
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Hydrogen Peroxide Prompted Lignification Affects Pathogenicity of Hemi-biotrophic Pathogen Bipolaris sorokiniana to Wheat

  • Poudel, Ajit (Institute of Agricultural Sciences, Banaras Hindu University) ;
  • Navathe, Sudhir (Institute of Agricultural Sciences, Banaras Hindu University) ;
  • Chand, Ramesh (Institute of Agricultural Sciences, Banaras Hindu University) ;
  • Mishra, Vinod K. (Institute of Agricultural Sciences, Banaras Hindu University) ;
  • Singh, Pawan K. (International Maize and Wheat Improvement Center (CIMMYT)) ;
  • Joshi, Arun K. (Borlaug Institute for South Asia (BISA))
  • Received : 2018.09.07
  • Accepted : 2019.04.22
  • Published : 2019.08.01
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Abstract

Spot blotch caused by Bipolaris sorokiniana has spread to more than 9 million ha of wheat in the warm, humid areas of the Eastern Gangetic Plains (EGP) of South Asia and is a disease of major concern in other similar wheat growing regions worldwide. Differential lignin content in resistant and susceptible genotypes and its association with free radicals such as hydrogen peroxide ($H_2O_2$), superoxide ($O_2{^-}$) and hydroxyl radical ($OH^-$) were studied after inoculation under field conditions for two consecutive years. $H_2O_2$ significantly influenced lignin content in flag leaves, whereas there was a negative correlation among lignin and $H_2O_2$ to the Area Under Disease Progress Curve (AUDPC). The production of $H_2O_2$ was higher in the resistant genotypes than susceptible ones. The $O_2{^-}$ and $OH^-$ positively correlated with AUDPC but negatively with lignin content. This study illustrates that $H_2O_2$ has a vital role in prompting lignification and thereby resistance to spot blotch in wheat. We used cluster analysis to separate the resistant and susceptible genotypes by phenotypic and biochemical traits. $H_2O_2$ associated lignin production significantly reduced the number of appressoria and penetration pegs. We visualized the effect of lignin in disease resistance using differential histochemical staining of tissue from resistant and susceptible genotypes, which shows the variable accumulation of hydrogen peroxide and lignin around penetration sites.

Keywords

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Fig. 1. Cluster analysis of 29 wheat genotypes on the basis their response to B. sorokiniana over two years in terms of (A) free radical production and lignin accumulation and, (B) various phenotypic traits such as AUDPC, LN, LS, DH, DM, PY, BM, and TKW. Clades highlighted with red lines are indicating clustering of susceptible genotypes whereas those of blue are resistant. (C) Plants from a resistant group; genotype Yangmai 6. (D) Plants showing typical symptoms of spot blotch seven days after inoculation on susceptible genotype Ciano T79.

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Fig. 2. Relative rates of resistance for the latent period, sporulation, fitness index, and AUDPC in resistant and susceptible genotypes. The relative rate of resistance in 29 wheat genotypes studied is calculated over the susceptible check Ciano T79.

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Fig. 3. Clustering of the correlated and non-correlated variables using the VARCLUS method in SAS v9.2 (A) Clustering of phenotypic traits and lignin content at three days. The components of the disease clustered together, indicating that enhanced resistance correlated to higher yield. (B) Clustering of biochemical traits indicating H2O2 is significantly affecting lignin production.

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Fig. 4. Accumulation of various reactive oxygen species and lignin at different time points in resistant and susceptible wheat genotypes in response to B. sorokiniana infection. (A) Accumulation of superoxide radicals (O2-). The plot is based on mean O2- accumulation in resistant and susceptible genotypes. LSD (0.05) for genotype = 0.0064 and for time = 0.0024. (B) Hydroxyl radical (OH-) production. The plot is based on mean OH- accumulation in resistant and susceptible genotypes. LSD (0.05) for genotype = 19.626 and for time = 7.2889. (C) Hydrogen peroxide (H2O2) production. The plot is based on mean H2O2 accumulation in resistant and susceptible genotypes. LSD (0.05) for genotype = 0.3143 and for time = 0.1167. (D) Lignin accumulation. The plot is based on mean H2O2 accumulation in resistant and susceptible genotypes. LSD (0.05) for genotype = 18.479 and for time = 6.8629. Data from two independent experiments are combined. ANOVA calculates the significant difference in the general linear model (GLM). Asterisks (*) show the significant difference as compared to time 0 DAI.

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Fig. 5. (A) Symptoms of B. sorokiniana on flag leaves over time; (B) Differential accumulation of H2O2 in response to B. sorokiniana visualized by DAB staining in flag leaves under time scan 0 to 14 days after infection (DAI). (C) Lignin deposition in flag leaves in response to B. sorokiniana as visualized by phloroglucinol-HCl staining in a longitudinal section of leaves from 0 to 14 DAI. (D) Lignin deposition in flag leaves in response to B. sorokiniana as visualized by phloroglucinol-HCl staining in the transverse section of leaves from 0 to 14 DAI. The upper panel represents resistant (R) genotype (Cv. Yangmai6) and the lower panel the susceptible (S) genotype (Cv. Ciano T79). Penetration points are marked by yellow arrowheads. Red line shown in the picture indicates size bar.

Table 1. Pedigree and spot blotch response of 29 wheat genotypes used in the experiment

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Table 3. Pearson’s correlation coefficients for disease, fitness parameters and relative rate of resistance for B. sorokiniana

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Table 4. Correlations among H2O2, OH-, O2-, and lignin at different time period before and after inoculation of spot blotch pathogen

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Table 2. Mean values of various disease related traits and relative rates of resistance in 29 wheat genotypes

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