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

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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.

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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).

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Authors and Affiliations

  1. Department of Food Science and Nutrition, Zhejiang University, Hangzhou, 310029, People’s Republic of China

    Xueyan Ren, Peng Wang, Feng Jiang, Huili Wang, Ting Yu & Xiaodong Zheng

  2. Department of Life Science, Shihezi University, Shihezi, 832003, People’s Republic of China

    Xueyan Ren & Qingjun Kong

  3. Department of Life Science, Tangshan Teacher’s College, Tangshan, 063000, People’s Republic of China

    Feng Jiang

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  1. Xueyan Ren
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  2. Qingjun Kong
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Correspondence to Xiaodong Zheng.

Electronic supplementary material

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11033_2010_169_MOESM1_ESM.doc

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

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

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)

Supplementary Table S1:Primers used in this study. (DOC 25 kb)

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Ren, X., Kong, Q., Wang, P. et al. Molecular cloning of a PR-5 like protein gene from cherry tomato and analysis of the response of this gene to abiotic stresses. Mol Biol Rep 38, 801–807 (2011). https://doi.org/10.1007/s11033-010-0169-0

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