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Molecular Biology Reports

, Volume 38, Issue 2, pp 801–807 | Cite as

Molecular cloning of a PR-5 like protein gene from cherry tomato and analysis of the response of this gene to abiotic stresses

  • Xueyan Ren
  • Qingjun Kong
  • Peng Wang
  • Feng Jiang
  • Huili Wang
  • Ting Yu
  • Xiaodong Zheng
Article

Abstract

LePR-5, a putative PR5 like protein gene was amplified from a cherry tomato (Lycopersicon esculentum), which encodes a precursor protein of 250 amino acid residues, and shares high degrees of homology with a number of other PR5 genes. Expression of LePR-5 in different tomato organs was analyzed with Semi-quantitative RT–PCR, showing that LePR-5 expressed at different levels in leaves, stems, roots, flowers and fruits. In addition, expression of LePR-5 under different abiotic stresses was carried out at different time points. Three of the four tested abiotic stimuli, ethophen, salicylic acid and methyl jasmonate, triggered a significant induction of LePR-5 after treatment. However, LePR-5 was weaker induced by abscisic acid than by others. The positive responses of LePR-5 to the three abiotic stimuli suggested that LePR-5 may play an important role in response to abiotic stresses, and it may also be involved in plant defense system against pathogens. In addition, different expression patterns between tomato fruit and seedling suggested that LePR-5 may play a distinctive role in the defensive system protecting tomato fruit and seedling.

Keywords

Molecular cloning Abiotic stress PR-5 like protein Cherry tomato Pathogenesis-related proteins 

Notes

Acknowledgments

This research was supported by the grants from the National Key Project of Scientific and Technical Supporting Programs Funded by Ministry of Science & Technology of China (2009badb9b09)and the National Natural Science Foundation Foundation of China (30771514) and the Ph.D. Programs Foundation of Ministry of Education of China (20090101120079).

Supplementary material

11033_2010_169_MOESM1_ESM.doc (33 kb)
Supplementary Fig. S1. Nucleotide and deduced amino acid sequences of LePR-5. The nucleic acid sequence of LePR-5 is presented on the top line, and the derived one-letter amino acid sequence is shown below. The stop code is marked with an asterisk. Primer sites used for cDNA clone, and RT–PCR were indicated by arrows with the respective primer names. Numbers on the left represents nucleotide and amino acid positions. The predicted N-terminal signal sequence is double underlined. (DOC 33 kb)
11033_2010_169_MOESM2_ESM.doc (41 kb)
Supplementary Fig. S2. Alignment of LePR-5 with N. tobacum (NtTLPx 03913), L. esculentum (LePR5: AY257487, LePR-5X : AY093595, LeNP24: M213461). Amino acid residues that are identical in all five sequences are shaded in dark, while well conserved residues are shaded in gray. The arrows indicate amino acid residues contained in the acidic cleft of NtTLP, LePR5, LePR-5, LePR-5x and LeNP24. Asterisk (*) show all 16 conserved cysteine residues among the aligned proteins. Suspension points (…) indicate sequence gaps to optimize the alignment. (DOC 41 kb)
11033_2010_169_MOESM3_ESM.doc (24 kb)
Supplementary Fig. S3. Phylogenetic tree of the thaumatin/PR5-like proteins was generated using the DNAman software. The amino acid sequences of PR5 used for construction of the tree are all deposited in the GenBank database under the following accession numbers (in parentheses): AsTLP-1 (L39774); AsTLP-2 (L39775); AsTLP-3 (L39776); AsTLP-4 (L39777); TaTLP (×97687.1); ZzPR5 (FJ550342); CaPR-5b (AJ010501); GmOLPb (AB370233); NtOLP (A16780); SnOLP (AF450276); StTLP (AF548357); GmOLPa (AB11625); AtPR5 (U83490); AtTLP (NM_106160); AtPR5-2 (NM_106161); CeTLP (AB031870); PaTLP (U32440); PpPsTL (AB036069). At, A. thaliana; Le, L. esculentum; Nt, N. tobacum; Gm, Glycine max, St, S. tuberosum; Sn, S. nigrum; Ca, Cicer arietinum; Zz, Zingiber zerumbet; Ta, Triticum aestivum; As, Avena sativa; Pa, Prunus avium; Pp, Pyrus pyrifolia; Ce, Cestrum elegans. (DOC 24 kb)
11033_2010_169_MOESM4_ESM.doc (24 kb)
Supplementary Table S1:Primers used in this study. (DOC 25 kb)

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Xueyan Ren
    • 1
    • 2
  • Qingjun Kong
    • 2
  • Peng Wang
    • 1
  • Feng Jiang
    • 1
    • 3
  • Huili Wang
    • 1
  • Ting Yu
    • 1
  • Xiaodong Zheng
    • 1
  1. 1.Department of Food Science and NutritionZhejiang UniversityHangzhouPeople’s Republic of China
  2. 2.Department of Life ScienceShihezi UniversityShiheziPeople’s Republic of China
  3. 3.Department of Life ScienceTangshan Teacher’s CollegeTangshanPeople’s Republic of China

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