Identification of putative phosphoproteins in wheat spikes induced by Fusarium graminearum

Lina Ding,Guoxing Yang,Ruiying Yang,Peng Li,Jun Cao,Yang Zhou

doi:10.1007/s00425-015-2441-y

Abstract

Main conclusionPhosphorylation and dephosphorylation events were initiated in wheat scab resistance. The putative FHB-responsive phosphoproteins are mainly involved in three functional groups and contain at least one tyrosine, serine, or threonine phosphorylation site.Fusarium head blight (FHB), caused by Fusarium graminearum, is a severe disease in wheat. Protein phosphorylation plays an important role in plant–pathogen interactions, however, a global analysis of protein phosphorylation in response to FHB infection remains to be explored. To study the effect of FHB on the phosphorylation state of wheat proteins, proteins extracted from spikes of a resistant wheat cultivar after 6 h of inoculation with F. graminearum or sterile H2O were separated by two-dimensional gel electrophoresis, and then the immunodetection of putative phosphoproteins was conducted by Western blotting using specific anti-phosphotyrosine antibody, anti-phosphothreonine antibody and anti-phosphoserine antibody. A total of 35 phosphorylated signals was detected and protein identities of 28 spots were determined. Functional categorization showed that the putative FHB-responsive phosphoproteins were mainly involved in defense/stress response, signal transduction, and metabolism. The phosphorylation status of proteins associated with signaling pathways mediated by salicylic acid, calcium ions, small GTPase, as well as with detoxification, reactive oxygen species scavenging, antimicrobial compound synthesis, and cell wall fortification was regulated in wheat spikes in response to F. graminearum infection. The present study reveals dynamics of wheat phosphoproteome in response to F. graminearum infection and suggests an important role of protein Ser/Thr/Tyr phosphorylation in fundamental mechanisms of wheat scab resistance.Electronic supplementary materialThe online version of this article (doi:10.1007/s00425-015-2441-y) contains supplementary material, which is available to authorized users.

Highlights

Fusarium head blight (FHB) or scab, caused by Fusarium graminearum, is a devastating disease in wheat (Triticum aestivum L.) and has been identified as a major factor limiting wheat production in many parts of world (Bai and Shannar 1994)
To study the effect of FHB on the phosphorylation state of wheat proteins, proteins extracted from spikes of a resistant wheat cultivar after 6 h of inoculation with F. graminearum or sterile H2O were separated by two-dimensional gel electrophoresis, and the immunodetection of putative phosphoproteins was conducted by Western blotting using specific anti-phosphotyrosine antibody, anti-phosphothreonine antibody and anti
One hundred micrograms of each protein extract of wheat spikes was separated by 2-DE and transferred to polyvinylidene fluoride (PVDF) membranes, phosphorylated signals were detected by immunostaining using anti-pTyr antibody, anti-pThr antibody and anti-pSer antibody respectively (Fig. 1)

Summary

Introduction

Fusarium head blight (FHB) or scab, caused by Fusarium graminearum, is a devastating disease in wheat (Triticum aestivum L.) and has been identified as a major factor limiting wheat production in many parts of world (Bai and Shannar 1994). Planta (2016) 243:719–731 graminearum is a semi-biotroph. During the early stages of infection of detached barley leaves by F. graminearum, hyphal growth occurs without host cell necrosis and the fungus behaves like a biotroph. During the infection of wheat spikes, F. graminearum produces cell-wall-degrading enzymes to facilitate penetration (Jaroszuk-Sciseł and Kurek 2012). The trichothecene mycotoxins produced by F. graminearum and F. culmorum (which are called FHB) are known to inhibit protein synthesis and may have a role in pathogenesis, leading to a reduction in grain yield and quality (Boenisch and Schafer 2011; Scherm et al 2013). There is an effect of chemical control, breeding for FHB-resistant cultivars is still the best means to control this disease (Kollers et al 2013; Lu et al 2013; Niwa et al 2014)

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