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Molecular characterization of a receptor-like protein kinase gene from pine (Pinus sylvestris L.)

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Abstract

We are developing molecular approaches to study the growth and development of woody plants. As part of our research efforts, we report the molecular cloning and characterization of PsRLK here, a cDNA from the conifer Scots pine (Pinus sylvestris L.) encoding a polypeptide similar to the receptor protein kinases described in angiosperms. A full-length clone was isolated from a cDNA library constructed with poly (A)+ enriched RNA prepared from germinating pine seeds. Characterization of the isolated sequence revealed that it contains multiple leucine-rich repeats in the N-terminal region and a characteristic Ser/Thr protein kinase domain in the C-terminal region. N- and C-terminal conserved domains are separated by a putative membrane spanning sequence. PsRLK protein is encoded by a single gene in the pine genome. A comparison of the pine sequence with the LRR-RLKs from Arabidopsis revealed that PsRLK is phylogenetically related to the LRR XI subfamily members. RT-PCR analyses of transcript abundance in pine tissues suggest that the gene expression pattern of PsRLK reflects the plant body formation programme, with increased levels during development of pine seedlings. The precise localization of PsRLK transcripts revealed that gene expression was restricted to specialized phloem cells suggesting a possible function of the putative receptor-like protein kinase in this particular vascular element.

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Abbreviations

EST:

Expressed sequence tag

LRR:

Leucine-rich repeat

RLK:

Receptor-like kinase

References

  • Allona I, Quinn M, Shoop I, Swope K, St Cyr S, Carlis J, Ried J, Retzel E, Campbell MM, Sederoff R, Whetten RW (1998) Analysis of xylem formation in pine by cDNA sequencing. Proc Natl Acad Sci USA 95:9693–9698

    Article  PubMed  CAS  Google Scholar 

  • Altschul SF, Gish W, Millar W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    Article  PubMed  CAS  Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1987) Current protocols in molecular biology. Green Publishing Associates and Wiley-Interscience, NewYork

    Google Scholar 

  • Ávila-Sáez C, Muñoz-Chapuli R, Plomion C, Frigerio J-M, Cánovas FM (2000) Two genes encoding distinct cytosolic glutamine synthetases are closely linked in the pine genome. FEBS Lett 477:237–243

    Article  PubMed  Google Scholar 

  • Ávila C, Cánovas FM. (2000) An improved and rapid protocol for the isolation of polyA+ from small amounts of Scots pine seedlings. Plant Mol Biol Rep 18:117–122

    Article  Google Scholar 

  • Ávila C, Suárez MF, Gómez-Maldonado J, Cánovas FM (2001) Spatial and temporal expression of two cytosolic glutamine synthetase genes in Scots pine: functional implications on nitrogen metabolism during early stages of conifer development. Plant J 25:93–102

    Article  PubMed  Google Scholar 

  • Baker B, Zambryski P, Staskawicz B, Dinesh-Kumar SP (1997) Signalling in plant–microbe interactions. Science 276:726–733

    Article  PubMed  CAS  Google Scholar 

  • Cánovas FM, Cantón FR, Gallardo F, García-Gutierrez A, de Vicente A (1991) Accumulation of glutamine synthetase during early development of maritime pine (Pinus pinaster) seedlings. Planta 185:372–378

    Article  Google Scholar 

  • Cantón FR, Suárez MF, Josè-Estanyol M, Cánovas FM (1999) Expression analysis of a cytosolic glutamine synthetase gene in cotyledons of Scots pine seedlings: developmental, light regulation and spatial distribution of specific transcripts. Plant Mol Biol 40:623–634

    Article  PubMed  Google Scholar 

  • Cantón FR, Le Provost G, Garcia V, Barré A, Frigerio J-M, Paiva J, Fevereiro P, Avila C, Mouret J-F, de Daruvar A, Cánovas FM, Plomion C (2003) Transcriptome analysis of wood formation in maritime pine. In: Espinel S, Barredo Y, Ritter E (eds) Sustainable forestry, wood products and biotechnology. DFA-AFA Press, Vitoria-Gasteiz, Spain, pp 333–347

    Google Scholar 

  • Clark SE, Williams RW, Meyerowitz EM (1997) The CLAVATA1 gene encodes a putative receptor kinase that controls shoot and floral meristem size in Arabidopsis. Cell 59:575–585

    Article  Google Scholar 

  • Clark SE (1997) Organ formation at the vegetative shoot meristem Department of Biology. Plant Cell 9:1067–1076

    Article  PubMed  CAS  Google Scholar 

  • Claros MG, Cánovas FM (1998) Rapid high quality RNA preparation from pine seedlings. Plant Mol Biol Rep 16:9–18

    Article  CAS  Google Scholar 

  • Deeken R, Kaldenhoff R (1997) Light-repressible receptor protein kinase: a novel photoregulated gene from Arabidopsis thaliana. Planta 202:497–486

    Article  Google Scholar 

  • Dellaporta SL, Wood J, Hicks JB (1983) Isolation of DNA from higher plants. Plant Mol Biol Rep 4:19–21

    Article  Google Scholar 

  • Fiers M, Golemiec E, Xu J, van der Geest L, Heidstra R, Stiekema W, Liu C-M (2005) The 14-amino acid CLV3, CLE19, and CLE40 peptides trigger consumption of the root meristem in Arabidopsis through a CLAVATA2-dependent pathway. Plant Cell 17:2542–2553

    Article  PubMed  CAS  Google Scholar 

  • Gómez-Maldonado J, Cánovas FM, Avila C (2004) Molecular analysis of the 5′ upstream region of a gibberellin-inducible cytosolic glutamine synthetase gene (GS1b) expressed in pine vascular tissue. Planta 218:1036–1045

    Article  PubMed  CAS  Google Scholar 

  • Hanks SK, Quinn AM (1991) Protein kinase catalytic domain sequence database: identification of conserved features of primary structure and classification of family members. Methods Enzymol 200:38–62

    PubMed  CAS  Google Scholar 

  • Hong SW, Jon JH, Kwak JM, Nam HG (1997) Identification of a receptor-like protein kinase gene rapidly induced by abscisic acid, dehydration, high salt, and cold treatments in Arabidopsis thaliana. Plant Physiol 113:1203–1212

    Article  PubMed  CAS  Google Scholar 

  • Higgins DG, Thompson JD, Gibson TJ (1996) Using CLUSTAL for multiple sequence alignments. Methods Enzymol 266:383–402

    PubMed  CAS  Google Scholar 

  • Hunter T (1995) Protein kinases and phosphatases: the ying and yang of protein phosphorylation and signaling. Cell 80:225–236

    Article  PubMed  CAS  Google Scholar 

  • Jeong S, Trotochaud AE, Clark SE (1999) The Arabidopsis CLAVATA2 gene encodes a receptor-like protein required for the stability of the CLAVATA1 receptor-like kinase. Plant Cell 11:1925–1933

    Article  PubMed  CAS  Google Scholar 

  • Kobe B, Deisenhofer J (1994) The leucine-rich repeat: a versatile binding motif. Trends Biochem Sci 1:415–421

    Article  Google Scholar 

  • Kobe B, Deisenhofer J (1995) Proteins with leucine-rich repeats. Curr Opin Struct Biol 5:409–416

    Article  PubMed  CAS  Google Scholar 

  • Kobe B, Kajava AV (2001) The leucine-rich repeat as a protein recognition motif. Curr Opin Struct Biol 11:725–732

    Article  PubMed  CAS  Google Scholar 

  • Langdale JA (1993) In situ hybridization. In: Freeling M, Walbot V (eds) The Maize Handbook. Springer, Berlin Heidelberg New York, pp 165–180

    Google Scholar 

  • Li J, Chory J (1997) A putative leucine-rich repeat receptor kinase involved in brassinosteroid signal transduction. Cell 90:929–938

    Article  PubMed  CAS  Google Scholar 

  • Li J, Wen JQ, Lease KA, Doke JT, Tax FE, Walker JC (2002) BAK1, an Arabidopsis LRR receptor-like protein kinase, interacts with BRI1 and modulates brassinosteroid signaling. Cell 110:213–222

    Article  PubMed  CAS  Google Scholar 

  • Muschietti J, Eyal Y, McCormick S (1998) Pollen tube localization implies role in pollen–pistil interactions for the tomato receptor-like protein-kinases LePRK1 and LePRK2. Plant Cell 10:319–330

    Article  PubMed  CAS  Google Scholar 

  • Pérez-Rodríguez J, Suárez MF, Heredia R, Avila C, Breton D, Trontin JF, Filonova L, Bozhkov P, von Arnold S, Harvengt L, Cánovas FM (2006) Expression patterns of two glutamine synthetase genes in zygotic and somatic pine embryos support specific roles in nitrogen metabolism during embryogenesis. New Phytol 169:35–44. DOI 10.1111/j.1469–8137.2005.01551.x

    Google Scholar 

  • Schmidt EDL, Guzzo F, Toonen MAJ, de Vries SC (1997) A leucine-rich repeat containing receptor-like kinase marks somatic plant cells competent to form embryos. Development 124:2049–2062

    PubMed  CAS  Google Scholar 

  • Shiu S-H, Blecker AB (2001) Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases. Proc Natl Acad Sci USA 98:10763–10768

    Article  PubMed  CAS  Google Scholar 

  • Shiu S-H, Blecker AB (2003) Expansion of the receptor-like kinase/Pelle gene family and receptor-like proteins in Arabidopsis. Plant Physiol 132:530–543

    Article  PubMed  CAS  Google Scholar 

  • Shiu S-H, Karlowski WM, Pan R, Tzeng Y-H, Mayer KFX, Li W-H (2004) Comparative analysis of the receptor-like kinase family in arabidopsis and rice. Plant Cell 16:1220–1234

    Article  PubMed  CAS  Google Scholar 

  • Song W, Wang G, Chen L, Kim H, Pi L, Holsten T, Gardner J, Wang B, Zhai W, Zhu L, Fauquet C, Ronald P (1995) A receptor kinase-like protein-encoded by the rice disease resistance gene, Xa21. Science 270:1084–1086

    Article  Google Scholar 

  • Suárez MF, Avila C, Gallardo F, Cantón FR, García-Gutiérrez A, Claros MG, Cánovas FM (2002) Molecular and enzymatic analysis of ammonium assimilation in woody plants. J Exp Bot 53:891–904

    Article  PubMed  Google Scholar 

  • Suzaki T, Sato M, Ashikari M, Miyoshi M, Nagato Y, Hirano H-Y (2004) The gene FLORAL ORGAN NUMBER1 regulates floral meristem size in rice and encodes a leucine-rich repeat receptor kinase orthologous to Arabidopsis CLAVATA1. Development 131:5649–5657

    Article  PubMed  CAS  Google Scholar 

  • Torii KU, Mitsukawa N, Oosumi T, Matsuura Y, Yokoyama R, Whittier RF, Komeda Y (1996) The Arabidopsis ERECTA gene encodes a putative receptor protein kinase with extracellular leucine-rich repeats. Plant Cell 8:735–746

    Article  PubMed  CAS  Google Scholar 

  • Trotochaud AE, Jeong S, Clark SE (2000) CLAVATA3, a multimeric ligand for the CLAVATA1 receptor-kinase. Science 289:613–617

    Article  PubMed  CAS  Google Scholar 

  • Van der Knaap E, Sauter M, Wilford R, Kende H (1996) Identification of a gibberellin induced receptor-like kinase in deep-water rice. Plant Physiol 112:1397–1397

    Article  PubMed  Google Scholar 

  • Walker JC (1993) Receptor-like protein kinase genes of Arabidopsis thaliana. Plant J 3:451–456

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank Remedios Crespillo for her excellent technical assistance. We are also grateful for the research facilities at the Molecular Biology Laboratory, Research Services, Universidad de Málaga. This work has been supported by Grants (PB98–1396 and BMC2003–04772) from Ministerio de Ciencia y Tecnología (Spain).

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Correspondence to Francisco M. Cánovas.

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Ávila, C., Pérez-Rodríguez, J. & Cánovas, F.M. Molecular characterization of a receptor-like protein kinase gene from pine (Pinus sylvestris L.). Planta 224, 12–19 (2006). https://doi.org/10.1007/s00425-005-0184-x

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  • DOI: https://doi.org/10.1007/s00425-005-0184-x

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