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. 2018 Aug 15;11(1):47.
doi: 10.1186/s12284-018-0240-3.

Proteomic analysis of the defense response to Magnaporthe oryzae in rice harboring the blast resistance gene Piz-t

Dagang Tian  1   2 , Liu Yang  3 , Zaijie Chen  1 , Ziqiang Chen  1 , Feng Wang  1 , Yuanchang Zhou  2 , Yuming Luo  4 , Liming Yang  5 , Songbiao Chen  6   7
Affiliations

Proteomic analysis of the defense response to Magnaporthe oryzae in rice harboring the blast resistance gene Piz-t

Dagang Tian et al. Rice (N Y). .

Abstract

Background: Rice blast (caused by Magnaporthe oryzae) is one of the most destructive diseases of rice. While many blast resistance (R) genes have been identified and deployed in rice cultivars, little is known about the R gene-mediated defense mechanism. We used a rice transgenic line harboring the resistance gene Piz-t to investigate the R gene-mediated resistance response to infection.

Results: We conducted comparative proteome profiling of the Piz-t transgenic Nipponbare line (NPB-Piz-t) and wild-type Nipponbare (NPB) inoculated with M. oryzae at 24, 48, 72 h post-inoculation (hpi) using isobaric tags for relative and absolute quantification (iTRAQ) analysis. Comparative analysis of the response of NPB-Piz-t to the avirulent isolate KJ201 and the virulent isolate RB22 identified 114 differentially expressed proteins (DEPs) between KJ201-inoculated NPB-Piz-t (KJ201-Piz-t) and mock-treated NPB-Piz-t (Mock-Piz-t), and 118 DEPs between RB22-inoculated NPB-Piz-t (RB22-Piz-t) and Mock-Piz-t. Among the DEPs, 56 occurred commonly in comparisons KJ201-Piz-t/Mock-Piz-t and RB22-Piz-t/Mock-Piz-t. In a comparison of the responses of NPB and NPB-Piz-t to isolate KJ201, 93 DEPs between KJ201-Piz-t and KJ201-NPB were identified. DEPs in comparisons KJ201-Piz-t/Mock-Piz-t, RB22-Piz-t/Mock-Piz-t and KJ201-Piz-t/KJ201-NPB contained a number of proteins that may be involved in rice response to pathogens, including pathogenesis-related (PR) proteins, hormonal regulation-related proteins, defense and stress response-related proteins, receptor-like kinase, and cytochrome P450. Comparative analysis further identified 7 common DEPs between the comparisons KJ201-Piz-t/KJ201-NPB and KJ201-Piz-t/RB22-Piz-t, including alcohol dehydrogenase I, receptor-like protein kinase, endochitinase, similar to rubisco large subunit, NADP-dependent malic enzyme, and two hypothetical proteins.

Conclusions: Our results provide a valuable resource for discovery of complex protein networks involved in the resistance response of rice to blast fungus.

Keywords: Piz-t; Proteomic analysis; Resistance gene; Rice blast disease; iTRAQ.

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Competing interests

The authors declare that they have no competing interests.

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Figures

Fig. 1
Fig. 1
Schematic representation of the experimental design. a The workflow for proteomic analysis of the defense response to M. oryzae in rice harboring the blast resistance Piz-t gene using iTRAQ 2D LC-MS/MS technology. b Phenotypes of the wild-type rice cultivar Nipponbare (NPB) and transgenic Nipponbare line harboring the Piz-t gene (NPB-Piz-t) inoculated with M. oryzae isolates KJ201 and RB22, respectively
Fig. 2
Fig. 2
qRT-PCR (a) and western blot (b) confirmation of the three differentially expressed proteins identified by iTRAQ
Fig. 3
Fig. 3
Venn diagrams of DEPs in NPB-Piz-t inoculated with M. oryzae isolates at 24 (a), 48 (b), and 72 hpi (c), respectively. 116: mock-treated NPB-Piz-t; 117: KJ201-inoculated NPB-Piz-t; 118: RB22-inoculated NPB-Piz-t; Up: upregulated proteins; Down: Downregulated proteins
Fig. 4
Fig. 4
GO analysis of DEPs in NPB-Piz-t in response to M. oryzae inoculation. a GO analysis of DEPs specific to comparison KJ201-Piz-t/Mock-Piz-t. b GO analysis of DEPs specific to comparison RB22-Piz-t/Mock-Piz-t. c GO analysis of common DEPs in comparisons KJ201-Piz-t/Mock-Piz-t and RB22-Piz-t/Mock-Piz-t. The results are summarized in three main categories: biological process, cellular component, and molecular function
Fig. 5
Fig. 5
KEGG classification of DEPs in NPB-Piz-t in response to M. oryzae inoculation. a KEGG classification of DEPs specific to comparison KJ201-Piz-t/Mock-Piz-t. b KEGG classification of DEPs specific to comparison RB22-Piz-t/Mock-Piz-t. c KEGG classification of common DEPs in comparisons KJ201-Piz-t/Mock-Piz-t and RB22-Piz-t/Mock-Piz-t
Fig. 6
Fig. 6
PPI networks of DEPs in NPB-Piz-t in response to M. oryzae inoculation. Response to KJ201: DEPs specific to comparison KJ201-Piz-t/Mock-Piz-t; Response to RB22: DEPs specific to comparison RB22-Piz-t/Mock-Piz-t; Co-Response: common DEPs in comparisons KJ201-Piz-t/Mock-Piz-t and RB22-Piz-t/Mock-Piz-t. P-values were calculated based on a -log scale. Fold change indicates the expression level of protein in the networks and are indicated in the white circle above each protein name
Fig. 7
Fig. 7
Expression patterns of DEPs may be involved in rice response to pathogens. a DEPs in NPB-Piz-t in response to M. oryzae inoculation. b DEPs in comparison between NPB-Piz-t and NPB in response to M. oryzae isolate KJ201. 114: KJ201-inoculated NPB; 116: mock-treated NPB-Piz-t; 117: KJ201-inoculated NPB-Piz-t; 118: RB22-inoculated NPB-Piz-t
Fig. 8
Fig. 8
PPI networks of DEPs in comparison between NPB-Piz-t and NPB in response to M. oryzae isolate KJ201. 24 h: 24 hpi; 48 h: 48 hpi; 72 h: 72 hpi
See this image and copyright information in PMC

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