Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 10 (1), 6

OsRLCK 57, OsRLCK107 and OsRLCK118 Positively Regulate Chitin- And PGN-Induced Immunity in Rice

Affiliations

OsRLCK 57, OsRLCK107 and OsRLCK118 Positively Regulate Chitin- And PGN-Induced Immunity in Rice

Zhangqun Li et al. Rice (N Y).

Abstract

Background: The receptor-like cytoplasmic kinase, OsRLCK176 has been reported to participate in both chitin- and PGN-induced immunity in rice. Here, we further researched the function of the homologous proteins, OsRLCK57, OsRLCK107 and OsRLCK118, in chitin- and PGN immunity in rice.

Findings: Silencing of OsRLCK57,OsRLCK107 and OsRLCK118 suppressed chitin- and PGN-induced immunity responses, including reactive oxygen species generation, defense gene expression. Furthermore, OsRLCK107 could interact with OsCERK1 in a MAMP induced way, which suggested a possible physiological relevance of OsRLCKs107 to OsCERK1 pathway.

Conclusions: OsRLCK57, OsRLCK107 and OsRLCK118, positively regulate chitin- and PGN- induced responses in rice, similar to that observed in OsRLCK176.

Keywords: Microbe-Associated Molecular Pattern; Pattern Triggered Immunity; Receptor-Like Cytoplasmic Kinases; Rice.

Figures

Fig. 1
Fig. 1
Characterization of homologous OsRLCK57,OsRLCK107 and OsRLCK118. a OsRLCK57, OsRLCK107, OsRLCK118 is highly homologous with OsRLCK176 and BIK1. The phylogenetic tree was generated by MEGA4. Full length amino acid sequences were selected to generate the bootstrap neighbor-joining phylogenetic tree. Bootstrap probabilities were obtained from 1000 replicates. Scale bar is indicated. b OsRLCK57, OsRLCK107 and OsRLCK118 is plasma membrane localized protein. Subcellular localization of OsRLCK57, OsRLCK107 and OsRLCK118 was visualized using confocal microscope after transiently transformed OsRLCK57-GFP, OsRLCK107-GFP and OsRLCK118-GFP into rice protoplasts. Chl indicated the chlorophyll autofluorescence. Three biological replicates were performed, similar results were obtained. Bar = 10 μm. c Expression identification of RLCKs in different RNAi lines. The relative expression levels OsRLCK57, OsRLCK107 and OsRLCK118 were examined by quantitative RT-PCR in OsRLCK57, OsRLCK107 and OsRLCK118 RNAi lines, respectively. The expression of OsRLCK176 and OsRLCK185 was also examined in these RNAi lines to exclude its effects. Empty vector (1301) transgenic callus cells were used as control. 57i-1 and 57i-14: RNAi lines of OsRLCK57. 107i-5 and 107i-6: RNAi lines of OsRLCK107. 118i-1 and 118i-2: RNAi lines of OsRLCK118. The data represents the mean ± standard deviation (SD) from three biological replicates
Fig. 2
Fig. 2
OsRLCK57, OsRLCK107 and OsRLCK118 are involved in chitin- and PGN-triggered immunity in rice. a RNAi silencing of OsRLCKs compromise the burst of ROS induced by chitin and PGN. OsRLCK57-RNAi, OsRLCK107-RNAi, OsRLCK118-RNAi or empty vector (1301) transgenic callus cells were treated with 100 μg/mL chitin, PGN or sterile water (for mock) for 120 min in dark before assayed. The data represents the mean ± standard deviation (SD) from three biological replicates. Statistically significant differences for OsRLCK-RNAi compared with the 1301 were calculated using Student’s t-test and indicated by asterisks (*P < 0.05). b RNAi silencing of OsRLCKs compromise the induction of PR genes by chitin and PGN. OsPR5, OsPR10, PAL and PBZ1 were determined by quantitative RT-PCR after 6 h treatments with 100 μg/mL chitin, PGN, LPS or sterile water (for mock) in dark. Empty vector (1301) transgenic callus cells were used as control. The data represents the mean ± standard deviation (SD) from three biological replicates. Statistically significant differences for OsRLCK-RNAi compared with the 1301 were calculated using Student’s t-test and indicated by asterisks (*P < 0.05)
Fig. 3
Fig. 3
OsRLCK107 interacts with OsCERK1. a OsRLCK107 interacts with OsCERK1 by BiFC. OsRLCKs-YN and OsCERK1-YC were co-expressed in rice protoplasts, the reconstituted YFP fluorescence was visualized using confocal microscope. Chl indicated the chlorophyll autofluorescence. Positive control: co-expression of bZIP63-YN and bZIP63-YC, negative control: co-expression of OsRLCKs-YN and blank-YC. Bar = 10 μm. b The interaction between OsRLCK107 with OsCERK1 is regulated by chitin and PGN. Protoplasts expressing OsRLCK107 with a C-terminal 3 × FLAG epitope tag were treated with 150 μg/mL chitin、PGN or sterile water (for mock) for 10 min. After treatment, protein was prepared for immunoprecipitation with anti-FLAG GEL (Sigma). The presence of OsCERK1 in the complex was analyzed by OsCERK1 antibody. Protein exacted from wide type (WT) protoplasts was used as negative control. Three biological replicates were performed and similar results were obtained

Similar articles

See all similar articles

Cited by 6 PubMed Central articles

See all "Cited by" articles

References

    1. Akamatsu A, Wong HL, Fujiwara M, Okuda J, Nishide K, Uno K, Imai K, Umemura K, Kawasaki T, Kawano Y, Shimamoto K. An OsCEBiP/OsCERK1-OsRacGEF1-OsRac1 Module is an Essential Early Component of Chitin-Induced Rice Immunity. Cell Host Microbe. 2013;13:465–476. doi: 10.1016/j.chom.2013.03.007. - DOI - PubMed
    1. Ao Y, Li Z, Feng D, Xiong F, Liu J, Li JF, Wang M, Wang J, Liu B, Wang HB. OsCERK1 and OsRLCK176 Play Important Roles in Peptidoglycan and Chitin Signaling in Rice Innate Immunity. Plant J. 2014;80:1072–1084. doi: 10.1111/tpj.12710. - DOI - PubMed
    1. Ausubel FM. Are Innate Immune Signaling Pathways in Plants and Animals Conserved? Nat. Immunol. 2005;6:973–979. doi: 10.1038/ni1253. - DOI - PubMed
    1. Cao Y, Liang Y, Tanaka K, Nguyen CT, Jedrzejczak RP, Joachimiak A, Stacey G (2014) The kinase LYK5 is a major chitin receptor in Arabidopsis and forms a chitin-induced complex with related kinase CERK1. eLife 3:e03766 - PMC - PubMed
    1. Kaku H, Nishizawa Y, Ishii-Minami N, Akimoto-Tomiyama C, Dohmae N, Takio K, Minami E, Shibuya N. Plant Cells Recognize Chitin Fragments for Defense Signaling Through A Plasma Membrane Receptor. Proc. Natl. Acad. Sci. U. S. A. 2006;103:11086–11091. doi: 10.1073/pnas.0508882103. - DOI - PMC - PubMed

LinkOut - more resources

Feedback