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TMT-based quantitative membrane proteomics reveals in-depth investigation of PRRs involved in the perception of MSP1 in rice leaves

ABSTRACT: Pathogen secreted pathogen-associated molecular patterns (PAMPs) play a key role in PAMPs triggered immunity (PTI) in plant for the recognition of pathogens. In rice, fewer PAMPs and their pattern recognition receptors (PRRs) have been characterized during rice-Magnaporthe oryzae interaction. Particularly, recent study was identified M. oryzae snodprot1 homolog known as MSP1, however, the molecular mechanism of MSP1 induced PTI is currently elusive. Therefore, we were generated the MSP1 overexpressed transgenic rice with their subcellular localization in the apoplastic (with signal sequence) and cytoplasmic (without signal sequence) regions, respectively, to examine the functional role of MSP1 in rice. Here, we employed a tandem mass tag (TMT)-based quantitative membrane proteomic analysis to decipher the potential interaction PRRs and MSP1-induced downstream signaling. This approach led to the identification of 8,033 proteins and sequential statistical analysis were identified 2,226 differentially modulated proteins. Of these, 20 plasma membrane localized receptor like kinases (RLKs) with the increased abundance in response to MSP1. Moreover, activation of proteins related to the protein degradation and modification, calcium signaling, transcription factor, redox, and MAPK signaling were characterized. Taken together, our results indicated that potential PRR candidates involved in response to blast disease and thus suggesting the overview mechanism of the MSP1-induced signaling in rice leaves.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Oryza Sativa (rice)

TISSUE(S): Leaf

SUBMITTER: Sun Tae Kim

LAB HEAD: Sun Tae Kim

PROVIDER: PXD032689 | Pride | 2022-08-12

REPOSITORIES: Pride

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TMT-based quantitative membrane proteomics identified PRRs potentially involved in the perception of MSP1 in rice leaves.

Min Cheol Woo CW Jang Jeong Woo JW Lee Gi Hyun GH Gupta Ravi R Yoon Jinmi J Park Hyun Ji HJ Cho Hye Sun HS Park Sang Ryeol SR Kwon Soon-Wook SW Cho Lae-Hyeon LH Jung Ki-Hong KH Kim Yu-Jin YJ Wang Yiming Y Kim Sun Tae ST

Journal of proteomics 20220730

Pathogen-associated molecular patterns (PAMPs) play a key role in triggering PAMPs triggered immunity (PTI) in plants. In the case of the rice-Magnaporthe oryzae pathosystem, fewer PAMPs and their pattern recognition receptors (PRRs) have been characterized. Recently, a M. oryzae snodprot1 homolog protein (MSP1) has been identified that functions as PAMP and triggering the PTI responses in rice. However, the molecular mechanism underlying MSP1-induced PTI is currently elusive. Therefore, we gene ...[more]

PMID: 35914717

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Project description:Rare sugars are monosaccharides that are rarely present in nature. One of the rare sugars, D-psicose, had a negative effect on shoot growth of rice. This negative effect to rice growth was observed previously with D-allose, but not observed in other tested rare sugars. To clarify the response to D-psicose in rice at the gene expression level, we performed a microarray analysis using the Agilent Rice Oligo Microarray (44k, custom-made; Agilent Technologies, Redwood City, CA, USA). As a result, treatment of D-psicose caused high induction of many defense-related genes including pathogenesis-related (PR) genes in rice. An Agilent Rice Oligo Microarray (44k, custom-made; Agilent Technologies, Redwood City, CA, USA) was used for the microarray analysis. By using the RNeasy plant mini kit (Qiagen, Hilden, Germany), total RNA was extracted from shoots of two-leaf stage rice plants that had been treated with 0.5 mM D-psicose or no sugar for 2 days. For each biological replicate, material from at least five rice plants was pooled to provide a single sample for RNA extraction. We performed 3 biological replicates for each treatment. All microarray procedures and data analyses were performed according to the manufacturer’s instructions. RNA integrity was checked using an Agilent 2100 Bioanalyzer (Agilent Technologies, Redwood City, CA, USA). Total RNA (400 ng) was labeled with Cy3 or Cy5 using an Agilent Low RNA Input Fluorescent Linear Amplification Kit (Agilent Technologies). Fluorescently labeled targets were hybridized to Agilent Rice Oligo Microarrays for 17 h at 65°C. After hybridization, wash processes were performed according to the manufacturer’s instructions, and hybridized microarrays were scanned using an Agilent Microarray Scanner (G2505B and software G2565BA; Agilent Technologies). Feature extraction software (version 9.1; Agilent Technologies) was used to delineate and measure the signal intensity of each spot in the array, and to normalize intensities. The background was measured around each spot as local background, calculated by the Feature Extraction software. Statistical data extraction processes were performed according to the manufacturer’s instructions.

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