A high-throughput evaluation system for Arabidopsis mutants for defense signaling

Premkumar, A.; Miya, Ayako; Hiratsuka, Kazuyuki; Kawakami, Naoto; Shibuya, Naoto

doi:10.5511/plantbiotechnology.23.459

Cited by 20 publications

(24 citation statements)

References 36 publications

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“…The values were the average of ROS generation observed for 10 individual seedlings. Although the WT seedlings generated ROS extensively after the treatment with N-acetylchitooctaose (GlcNAc) 8 , as reported (14), the seedlings of the cerk1 mutants, cerk1-1 and cerk1-2, did not.…”

Section: Identification Of the Chitin Elicitor Receptor Kinase 1 (Cermentioning

confidence: 95%

“…Plant Growth and Elicitor Treatment. Arabidopsis seeds were surface-sterilized and germinated on Petri plates containing MGRL nutrients supplemented with 0.1% agarose and 1% sucrose (1% glucose in the case of LPS treatment), as reported (14). Seedlings were grown for 8-10 days Ͻ16/8-h light/dark cycle at 22°C (light) or 15°C (dark) and used for elicitor treatment.…”

Section: Methodsmentioning

confidence: 99%

“…We and others reported that chitin oligosaccharide elicitors induce the expression of various defense-related genes in both suspension-cultured rice cells and Arabidopsis seedlings (14,21,22). To investigate whether this response is impaired in cerk1-1, we performed RT-PCR experiments (Fig.…”

Section: Genome-wide Gene Response Induced By the Chitin Elicitor Wasmentioning

confidence: 99%

“…To see whether such a deficiency in elicitor response is specific to chitin oligosaccharide, the responses of WT as well as mutant seedlings to another potent elicitor, bacterial lipopolysaccharide (LPS) from Pseudomonas aeruginosa (14,15), was examined. As shown in Fig.…”

Section: Identification Of the Chitin Elicitor Receptor Kinase 1 (Cermentioning

confidence: 99%

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CERK1, a LysM receptor kinase, is essential for chitin elicitor signaling in Arabidopsis

Miya

Premkumar

Shinya

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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Chitin is a major component of fungal cell walls and serves as a microbe-associated molecular pattern (MAMP) for the detection of various potential pathogens in innate immune systems of both plants and animals. We recently showed that chitin elicitor-binding protein (CEBiP), plasma membrane glycoprotein with LysM motifs, functions as a cell surface receptor for chitin elicitor in rice. The predicted structure of CEBiP does not contain any intracellular domains, suggesting that an additional component(s) is required for signaling through the plasma membrane into the cytoplasm. Here, we identified a receptor-like kinase, designated CERK1, which is essential for chitin elicitor signaling in Arabidopsis. The KO mutants for CERK1 completely lost the ability to respond to the chitin elicitor, including MAPK activation, reactive oxygen species generation, and gene expression. Disease resistance of the KO mutant against an incompatible fungus, Alternaria brassicicola, was partly impaired. Complementation with the WT CERK1 gene showed cerk1 mutations were responsible for the mutant phenotypes. CERK1 is a plasma membrane protein containing three LysM motifs in the extracellular domain and an intracellular Ser/Thr kinase domain with autophosphorylation/myelin basic protein kinase activity, suggesting that CERK1 plays a critical role in fungal MAMP perception in plants.plant immunity ͉ host-pathogen interaction ͉ N-acetylchitooligosaccharide P lants trigger various defense reactions against invading pathogens upon perception of so-called microbe-associated molecular patterns [MAMPs; also known as pathogenassociated molecular patterns (PAMPs)]. MAMP recognition has been implicated to play a major role in the nonhost or basal resistance that makes most plants immune to most potential pathogens (1, 2). In the course of coevolution between hosts and parasites, it seems that pathogens developed various virulence factors to overcome MAMP-mediated defense responses, whereas plants evolved resistance-gene-mediated defense system to detect such factors (3-5). Recent studies also revealed the presence of a striking similarity in the MAMP-mediated immunity between plants and animals (2, 6).Despite the importance of MAMP-mediated immunity in plants, molecular machineries involved in the perception of MAMPs and their signal transduction have been poorly understood. So far, only the receptor molecules for two bacterial MAMPs, flg22 and EF-Tu, have been identified (7,8). Both of these receptors, FLS2 and EFR, are receptor kinases with a leucine-rich repeat in the extracellular domain. However, much less information is available for the perception of fungal MAMPs. Chitin and its fragments, chitin oligosaccharides or N-acetylchitooligosaccharides, are typical fungal MAMPs that trigger various defense responses in both monocots and dicots, indicating the presence of a conserved machinery to perceive these oligosaccharides in a wide range of plant species (9, 10). A recent finding that chitin also induces immune responses in mice (11) suggest...

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Section: Identification Of the Chitin Elicitor Receptor Kinase 1 (Cermentioning

confidence: 95%

Section: Methodsmentioning

confidence: 99%

Section: Genome-wide Gene Response Induced By the Chitin Elicitor Wasmentioning

confidence: 99%

Section: Identification Of the Chitin Elicitor Receptor Kinase 1 (Cermentioning

confidence: 99%

See 2 more Smart Citations

CERK1, a LysM receptor kinase, is essential for chitin elicitor signaling in Arabidopsis

Miya

Premkumar

Shinya

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

1,234

1,067

View full text Add to dashboard Cite

show abstract

“…In addition, the use of a BY-2 cell system would enable us to conduct an assay protocol with multi-well plates for the large-scale experiments (Albert et al 2006;Narusaka et al 2006). On the other hand, unlike the previously reported methods using a sophisticated detection system, the protocol described in this study only requires a conventional luminometer, and may also be suitable to small-scale experiments.…”

Section: Discussionmentioning

confidence: 99%

Non-destructive bioluminescence detection system for monitoring defense gene expression in tobacco BY-2 cells

Watakabe

Ono

Tanaka

et al. 2011

Plant Biotechnology

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The RNA-binding protein FPA regulates flg22-triggered defense responses and transcription factor activity by alternative polyadenylation

Lyons

Iwase

Gänsewig

et al. 2013

Sci Rep

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RNA-binding proteins (RBPs) play an important role in plant host-microbe interactions. In this study, we show that the plant RBP known as FPA, which regulates 3′-end mRNA polyadenylation, negatively regulates basal resistance to bacterial pathogen Pseudomonas syringae in Arabidopsis. A custom microarray analysis reveals that flg22, a peptide derived from bacterial flagellins, induces expression of alternatively polyadenylated isoforms of mRNA encoding the defence-related transcriptional repressor ETHYLENE RESPONSE FACTOR 4 (ERF4), which is regulated by FPA. Flg22 induces expression of a novel isoform of ERF4 that lacks the ERF-associated amphiphilic repression (EAR) motif, while FPA inhibits this induction. The EAR-lacking isoform of ERF4 acts as a transcriptional activator in vivo and suppresses the flg22-dependent reactive oxygen species burst. We propose that FPA controls use of proximal polyadenylation sites of ERF4, which quantitatively limit the defence response output.

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A high-throughput evaluation system for Arabidopsis mutants for defense signaling

Cited by 20 publications

References 36 publications

CERK1, a LysM receptor kinase, is essential for chitin elicitor signaling in Arabidopsis

CERK1, a LysM receptor kinase, is essential for chitin elicitor signaling in Arabidopsis

Non-destructive bioluminescence detection system for monitoring defense gene expression in tobacco BY-2 cells

The RNA-binding protein FPA regulates flg22-triggered defense responses and transcription factor activity by alternative polyadenylation

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