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Cloning and expression analysis of Rsk in Brassica napus induced by Sclerotinia sclerotiorum

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Abstract

Based on the significant differences in the resistance to Sclerotinia sclerotiorum between two Brassica napus cultivars, ZhongR888 and Zhongyou821, near isogenic lines (NILs) were established. The resistance differences between Zhongyou821 and BC5 progeny (the fifth generation from a cross between Zhongyou821 and ZhongR888 with Zhongyou821 as successive backcross parent) was displayed by cDNA-AFLP and differential transcripts-derived fragments (TDFs). The full-length cDNA of 1,707 bp with 1,323 bp open reading frame (ORF, open box), named Rsk was identified by RT-RACE from the resistance related fragment 40-2 in Zhongyou821. It encoded a protein of 440 amino acid residues, which contained a putative extracellular domain, a transmembrane domain, and an intracellular domain. Analysis of the deduced amino acid sequence of Rsk revealed that it had high homology to Arabidopsis thaliana kinase involved in protein binding, and had a conserved region of LRR-RI, indicating it might be a member of leucine-rich repeats, ribonuclease inhibitor-like subfamily. DNA sequences of 1,685 bp and 1,703 bp without an intron were also identified from Zhongyou821 and BC5, respectively. Expression analysis showed that Rsk might play a role in disease resistance pathways.

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References

  • Bachem CWB, Visser RGF (1998) Transcript imaging with cDNA-AFLP: a step-by-step protocol. Plant Mol Biol Rep 16:157–173

    Article  CAS  Google Scholar 

  • Bachem CWB, Van de Hoeven RS, De Bruijn SM, Vreugdenhil D, Zabeau M, Visser RGF (1996) Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: analysis of gene expression during potato tuber development. Plant J 9:745–753

    Article  PubMed  CAS  Google Scholar 

  • Barua UM, Chalmers KJ, Hackett CA, Thomas WTB, Powell W, Waugh R (1993) Identification of RAPD markers linked to a Rhynchosporium secalis resistance locus in barley using near-isogenic lines and bulked segregant analysis. Heredity 71:177–184

    Article  PubMed  CAS  Google Scholar 

  • Bolton MD, Thomma BPHJ, Nelson BD (2006) Sclerotinia sclerotiorum (Lib.) de Bary: biology and molecular traits of a cosmopolitan pathogen. Mol Plant Pathol 7:1–16

    Article  PubMed  CAS  Google Scholar 

  • Bom M, Boland GL (2000) Evaluation of disease forecasting variables for Sclerotinia stem rot (Sclerotinia sclerotiorum) of canola. Can J Plant Sci 80:889–898

    Article  Google Scholar 

  • Burke JM, Rieseberg LH (2003) Fitness effects of transgenic disease resistance in sunflowers. Science 300:1250

    Article  PubMed  CAS  Google Scholar 

  • Dellagi A, Birch PRJ, Heilbronn J, Lyon GD, Toth IK (2000) cDNA-AFLP analysis of differential gene expression in the bacterial plant pathogen Erwinia carotovora. Microbiology 146:165–171

    PubMed  CAS  Google Scholar 

  • Dias BBA, Cunha WG, Morais LS, Vianna GR, Rech EL, de Capdeville G, Aragão FJL (2006) Expression of an oxalate decarboxylase gene from Flammulina sp. in transgenic lettuce (Lactuca sativa) plants and resistance to Sclerotinia sclerotiorum. Plant Pathol 55:187–193

    Article  CAS  Google Scholar 

  • Durrant WE, Rowland O, Piedras P, Hammond-Kosack KE, Jones JDG (2000) cDNA-AFLP reveals a striking overlap in race-specific resistance and wound response gene expression profiles. Plant Cell 12:963–977

    Article  PubMed  CAS  Google Scholar 

  • Gan L, Wu XL, Jin L, Feng SQ, Chen CL, Tang H (2002) The establishment of Sclerotinia Sclerotiorum resistant near isogenic lines. J Wuhan Univ (Natural Science Edition) 6:761–764

    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

    Article  PubMed  CAS  Google Scholar 

  • He KY, Yi B, Fu TD, Tu JX (2005) Genetic analysis of resistance to Sclerotinia sclerotiorum in Brassica napus L. Acta Agron Sinica 31:1495–1499

    Google Scholar 

  • Hu BC, Rimmer SR (1989) Preliminary study of artificial inoculation for resistance (tolerance) to Sclerotinia Sclerotiorum in rapeseed using detached leaves. J Anhui Agric Sci 3:56–58

    Google Scholar 

  • Kim YC, Kim SY, Paek KH, Choi D, Park JM (2006) Suppression of CaCYP1, a novel cytochrome P450 gene, compromises the basal pathogen defense response of pepper plants. Biochem Biophys Res Commun 345:638–645

    Article  PubMed  CAS  Google Scholar 

  • Lawrence GJ, Finnegan EJ, Ayliffe MA, Ellis JG (1995) The L6 gene for flax rust resistance is related to the Arabidopsis bacterial resistance gene RPS2 and the tobacco viral resistance gene N. Plant Cell 7:1195–1206

    Article  PubMed  CAS  Google Scholar 

  • Li QS, McCartney HA, Heran A, Hu BC, Chen FX, Hou SM, Wu XJ, Fei WX (2000) Study on infection conditions of oilseed rape infected by Scterotinia stem rot Erotinia stem rot. J Anhui Agric Sci 28:314–315

    CAS  Google Scholar 

  • Lindberg RA, Quinn AM, Hunter T (1992) Dual-specificity protein kinases: will any hydroxyl do? Trends Biochem Sci 17:114–119

    Article  PubMed  CAS  Google Scholar 

  • Liu GZ, Pi LY, Walker JC, Ronald PC, Song WY (2002) Biochemical characterization of the kinase domain of the rice disease resistance receptor-like kinase Xa21. J Biol Chem 277:20264–20269

    Article  PubMed  CAS  Google Scholar 

  • Liu RH, Zhao JW, Xiao Y, Meng JL (2005a) Identification of prior candidate genes for Sclerotinia local resistance in Brassica napus using Arabidopsis cDNA microarray and Brassica-Arabidopsis comparative mapping. Sci China C Life Sci 48:460–470

    Article  PubMed  CAS  Google Scholar 

  • Liu S, Wang H, Zhang J, Fitt BDL, Xu Z, Evans N, Liu Y, Yang W, Guo X (2005b) In vitro mutation and selection of doubled-haploid Brassica napus lines with improved resistance to Sclerotinia sclerotiorum. Plant Cell Rep 24:133–144

    Article  PubMed  CAS  Google Scholar 

  • Livingstone DM, Hampton JL, Phipps PM, Grabau EA (2005) Enhancing resistance to Sclerotinia minor in Peanut by expressing a barley oxalate oxidase gene. Plant Physiol 137:1354–1362

    Article  PubMed  CAS  Google Scholar 

  • Mackill DJ, Boman JM (1992) Inheritance of blast resistance in near-isogenic lines of rice. Phytopathology 82:746–749

    Article  Google Scholar 

  • Ohmori T, Murata M, Motoyoshi F (1998) Characterization of disease resistance gene-like sequences in near-isogenic lines of tomato. Theor Appl Genet 96:331–338

    Article  CAS  Google Scholar 

  • Qin L, Prins P, Jones JT, Popeijus H, Smant G, Bakker J, Helder J (2001) GenEST, a powerful bidirectional link between cDNA sequence data and gene expression profiles generated by cDNA-AFLP. Nucleic Acids Res 29:1616–1622

    Article  PubMed  CAS  Google Scholar 

  • Shamrai SN (2003) Plant resistance genes: molecular and genetic organization, function and evolution. Zh Obshch Biol 64:195–214

    PubMed  CAS  Google Scholar 

  • Song WY, Wang GL, Chen LL, Kim HS, Pi LY, Holsten T, Gardner J, Wang B, Zhai WX, Zhu LH, Fauquet C, Ronald P (1995) A receptor kinase-like protein encoded by the rice disease resistance gene, Xa21. Science 270:1804–1806

    Article  PubMed  CAS  Google Scholar 

  • Tang X, Frederick RD, Zhou J, Halterman DA, Jia Y, Martin GB (1996) Initiation of plant disease resistance by physical interaction of AvrPto and Pto kinase. 274:2060–2063

  • Walker JC (1994) Structure and function of the receptor-like protein kinases of higher plants. Plant Mol Biol 26:1599–1609

    Article  PubMed  CAS  Google Scholar 

  • Wang Z, Xian J (2004) Brassinosteroid signal transduction-choices of signals and receptors. Trends Plant Sci 9:91–96

    Article  PubMed  CAS  Google Scholar 

  • Wang ZM, Mackill DJ, Bonman JM (1989) Inheritance of partial resistance to blast in Indica rice cultivars. Corp science 29:848–853

    Article  Google Scholar 

  • Yang B, Srivastava S, Deyholos MK, Kav NNV (2007) Transcriptional profiling of canola (Brassica napus L.) responses to the fungal pathogen Sclerotinia sclerotiorum. Plant Sci 173:156–171

    Article  CAS  Google Scholar 

  • Yoshimura S, Yamanouchi U, Katayose Y, Toki S, Wang Z-X, Kono I, Kurata N, Yano M, Iwata N, Sasaki T (1998) Expression of Xa1, a bacterial blight-resistance gene in rice, is induced by bacterial inoculation. Proc Natl Acad Sci USA 95(4):1663–1668

    Article  PubMed  CAS  Google Scholar 

  • Yu YG, Buss GR, Saghai Maroof MA (1996) Isolation of a superfamily of candidate disease-resistance genes in soybean based on a conserved nucleotide-binding site. Proc Natl Acad Sci USA 93:11751–11756

    Article  PubMed  CAS  Google Scholar 

  • Zhang LN, Niu JS, Yu L (2005) Expression analysis of wheat TaMlo-A1c gene by Semi-QRT-PCR. Acta Botanica Boreali-occidentalia Sinica 25:1368–1371

    CAS  Google Scholar 

Download references

Acknowledgments

We thank Prof. Zaiyun Li for revising this paper, and thank Jiuwei Xu for help and suggestions concerning these investigations. This study was supported by the National Nature Science Foundation of China (grant no. 30170600) and Hubei Nature Science Foundation (grant no. 2001ABD113).

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Correspondence to Li Gan.

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Communicated by L. Kleczkowski.

Supported by the National Nature Science Foundation of China (30170600) and Hubei Nature Science Foundation (2001ABD113).

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Zhang, Y., Hu, Cx., Zhang, Cg. et al. Cloning and expression analysis of Rsk in Brassica napus induced by Sclerotinia sclerotiorum . Acta Physiol Plant 33, 1277–1283 (2011). https://doi.org/10.1007/s11738-010-0658-8

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  • DOI: https://doi.org/10.1007/s11738-010-0658-8

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