Isolation of novel rice (Oryza sativa L.) multiple stress responsive MAP kinase gene, OsMSRMK2, whose mRNA accumulates rapidly in response to environmental cues

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Abstract

In search for components of MAPK (mitogen-activated protein kinase) cascades in rice (Oryza sativa L. cv. Nipponbare), we identified a single copy gene called OsMSRMK2 from jasmonic acid (JA) treated rice seedling leaf cDNA library. This gene has a conserved protein kinase domain, including a MAPK family signature, and encodes a 369 amino acid polypeptide with a predicted molecular mass of 42995.43 and a pI of 5.48. OsMSRMK2 did not show constitutive expression in leaves and was induced within 15 min in response to wounding by cut. Using in vitro system, we show that the expression of OsMSRMK2 mRNA was potently enhanced within 15 min by signalling molecules, protein phosphatase inhibitors, ultraviolet irradiation, fungal elicitor, heavy metals, high salt and sucrose, and drought. OsMSRMK2 expression was further modulated by co-application of JA, salicylic acid, and ethylene and required de novo synthesized protein factor(s) in its transient regulation. Moreover, high (37 °C) and low temperatures (12 °C) and environmental pollutants—ozone and sulfur dioxide—differentially regulate the OsMSRMK2 mRNA accumulation in leaves of intact plants. Present results demonstrating dramatic transcriptional and transient regulation of the OsMSRMK2 expression by diverse biotic/abiotic stresses, a first report for any rice (or plant) MAPK to date, suggest a role for OsMSRMK2 in rice defense/stress response pathways.

Section snippets

Materials and methods

Plant material and in vitro system. Rice (O. sativa L. cv. Nipponbare) was grown under white fluorescent light (wavelength 390–500 nm, 150μmolm−2s−1, 12 h light period/day) at 25 °C and 70% relative humidity as described previously [5], [6], [7]. For in vitro experiments, the middle portions of fully expanded leaves (approximately 2 cm long segments cut with sterile scissor) from two-week-old seedlings were used for all treatments under continuous light (150μmolm−2s−1). Leaf segments placed in open

Identification of the OsMSRMK2 gene

To isolate rice genes involved in the early steps of rice plant defense/stress response pathways, we embarked on isolating specific genes induced in response to JA, a crucial endogenous signalling molecule [1], [2], [4], [5], [6], using differential cloning and identified a novel MAPK gene, OsMSRMK2 from japonica-type rice (cv. Nipponbare) seedling leaves. The OsMSRMK2 cDNA is 1356-bp long and contains an open reading frame of 1110 nucleotides capable of encoding a 369 amino acid polypeptide

Concluding remarks

Results described and discussions thereof clearly show that OsMSRMK2: (a) is a novel member of the MAPKs from japonica-type rice, (b) is induced by and thus perceives global signalling molecules and biotic and abiotic stresses on rice leaves and hence triggered in response to rice defense/stress cues, (c) is negatively regulated by a de novo synthesized protein factor, and very importantly (d) gives expression patterns at the transcript level that have not been seen (or shown) in dicots.

Acknowledgements

R.R. is a close collaborator of the RLABB and presently works at AIST supported by the Japan Society for Promotion of Science. We thank Drs. A. Kubo and H. Saji (National Institute of Environmental Studies, Tsukuba, Japan) for use of the ozone and sulfur dioxide fumigation facilities.

References (36)

  • N Shibuya et al.

    Oligosaccharide signalling for defence responses in plant

    Physiol. Mol. Plant Pathol.

    (2001)
  • B.J Feys et al.

    Interplays of signaling pathways in plant disease resistance

    Trends Genet.

    (2000)
  • S.A Goff et al.

    A draft sequence of the rice genome (Oryza sativa L. ssp. japonica)

    Science

    (2002)
  • R Rakwal et al.

    Separation of proteins from stressed rice (Oryza sativa L.) leaf tissues by two-dimensional polyacrylamide gel electrophoresis: induction of pathogenesis related and cellular protectant proteins by jasmonic acid, ultraviolet irradiation and copper chloride

    Electrophoresis

    (1999)
  • J.W Widmann et al.

    Mitogen-activated protein kinase: conservation of a three-kinase module from yeast to human

    Physiol. Rev.

    (1999)
  • H.J Schaeffer et al.

    Mitogen-activated protein kinases: specific messages from ubiquitous messengers

    Mol. Cell. Biol.

    (1999)
  • S Usami et al.

    Cutting activates a 46-kDa protein kinase in plants

    Proc. Natl. Acad. Sci. USA

    (1995)
  • S Seo et al.

    Tobacco MAP kinase: a possible mediator in wound signal transduction pathways

    Science

    (1995)
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    These authors contributed equally to this work.

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