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Isolation and characterization of a gene encoding a polyethylene glycol-induced cysteine protease in common wheat

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Abstract

Plant cysteine protease (CP) genes are induced by abiotic stresses such as drought, yet their functions remain largely unknown. We isolated the full-length cDNA encoding a Triticum aestivum CP gene, designated TaCP, from wheat by the rapid amplification of cDNA ends (RACE) method. Sequence analysis revealed that TaCP contains an open reading frame encoding a protein of 362 amino acids, which is 96% identical to barley cysteine protease HvSF42. The TaCP transcript level in wheat seedlings was upregulated during polyethylene glycol (PEG) stress, with a peak appearing around 12 h after treatment. TaCP expression level increased rapidly with NaCl treatment at 48 h. TaCP responded strongly to low temperature (4°C) treatment from 1 h post-treatment and reached a peak of about 40-fold at 72 h. However, it showed only a very slight response to abscisic acid (ABA). More than one copy of TaCP was present in each of the three genomes of hexaploid wheat and its diploid donors. TaCP fused with green fluorescent protein (GFP) was located in the plasma membrane of onion epidermis cells. Transgenic Arabidopsis plants overexpressing TaCP showed stronger drought tolerance and higher CP activity under water-stressed conditions than wild-type Arabidopsis plants. The results suggest that TaCP plays a role in tolerance to water deficit.

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Abbreviations

ABA:

abscisic acid

CaMV:

cauliflower mosaic virus

CP:

cysteine protease

EST:

expressed sequence tag

GFP:

green fluorescent protein

ORF:

open reading frame

PEG:

polyethylene glycol

pI:

isoelectric point

PVP:

polyvinylpyrrolidone

RACE:

rapid amplification of cDNA ends

UTR:

untranslated region

References

  • Bechtold N, Ellis J and Pelletier G 1993 In planta Agrobacteriummediated gene transfer by infiltration of Arabidopsis thaliana plants; C. R. Acad. Sci. Paris 316 1194–1199

    CAS  Google Scholar 

  • Bradfold M M 1976 A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding; Anal. Biochem. 72 248–254

    Article  Google Scholar 

  • Bray E A 1997 Plant responses to water deficit; Trends Plant Sci. 2 48–54

    Article  Google Scholar 

  • Buchanan W V 1997 The molecular biology of leaf senescence; J. Exp. Bot. 48 181–199

    Article  Google Scholar 

  • Cercos M, Santamaria S and Carbonell J 1999 Cloning and characterization of TPE4A, a thiol-protease gene induced during ovary senescence and seed germination in pea; Plant Physiol. 119 1341–1348

    Article  CAS  PubMed  Google Scholar 

  • Church G M and Gilbert W 1984 Genomic sequencing; Proc. Natl. Acad. Sci. USA 81 1991–1995

    Article  CAS  PubMed  Google Scholar 

  • Dellaporta S L, Wood J and Hicks J B 1983 A plant DNA minipreparation version II; Plant Mol. Biol. Rep. 1 19–21

    Article  CAS  Google Scholar 

  • Domoto C, Watanabe H, Abe M, Abe K and Arai S 1995 Isolation and characterization of two distinct cDNA clones encoding corn seed cysteine proteases; Biochim. Biophys. Acta 1263 241–244

    PubMed  Google Scholar 

  • Drake R, John I, Farrell A, Cooper W, Schuch W and Grierson D 1996 Isolation and analysis of cDNA encoding tomato cysteine protease expressed during leaf senescence; Plant Mol. Biol. 30 755–767

    Article  CAS  PubMed  Google Scholar 

  • Guan H C, Lin T H, Mee N Y, Ruey H L, Yih J H, Ming C C and Shu C G C 2002 Molecular characterization of a senescence-associated gene encoding cysteine proteinase and its gene expression during leaf senescence in sweet tomato; Plant Cell Physiol. 43 984–991

    Article  Google Scholar 

  • Guerrero F D, Jones J T and Mullet J E 1990 Turgor-responsive gene transcription and RNA levels increase rapidly when pea shoots are wilted. Sequence and expression of three inducible genes; Plant Mol. Biol. 15 11–26

    Article  CAS  PubMed  Google Scholar 

  • Harrak H, Azelmat S, Baker E N and Tabaeizadeh Z 2001 Isolation and characterization of a gene encoding a drought-induced cysteine protease in tomato (Lycopersicon esculentum); Genome 44 368–374

    Article  CAS  PubMed  Google Scholar 

  • Ingram J and Bartels D 1996 The molecular basis of dehydration tolerance in plants; Annu. Rev. Plant Physiol. Plant Mol. Biol. 47 377–403

    Article  CAS  PubMed  Google Scholar 

  • Jones J T and Mullet J E 1995 A salt-inducible and dehydration-inducible pea gene, Cyp15a, encodes a cell-wall protein with sequence similarity to cysteine proteases; Plant Mol. Biol. 28 1055–1065

    Article  CAS  PubMed  Google Scholar 

  • Kazuo S and Kazuo Y S 1997 Gene expression and signal transduction in water-stress response; Plant Physiol. 115 327–334

    Article  Google Scholar 

  • Koizumi M, Yamaguchi-Shinozaki K, Tsuji H and Shinozaki K 1993 Structure and expression of two genes that encode distinct drought inducible cysteine proteases in Arabidopsis thaliana; Gene 129 175–182

    Article  CAS  PubMed  Google Scholar 

  • Korthout H A A J, Berecki G, Bruin W, van Duijn B and Wang M 2000 The presence and subcellular localization of caspase 3-like proteinases in plant cells; FEBS Lett. 475 139–144

    Article  CAS  PubMed  Google Scholar 

  • Kusaka K, Tada Y, Shigemi T, Sakamoto M, Nakayashiki H, Tosa Y and Mayama S 2004 Coordinate involvement of cysteine protease and nuclease in the executive phase of plant apoptosis; FEBS Lett. 578 363–367

    Article  CAS  PubMed  Google Scholar 

  • Li Q, Bettany A J, Donnison I, Griffiths C M, Thomas H and Scott I M 2000 Characterisation of a cysteine protease cDNA from lolium multiform leaves and its expression during senescene and cytokinin treatment; Biochim. Biophys. Acta 1492 233–236

    CAS  PubMed  Google Scholar 

  • Lid S E, Gruis D, Jung R, Lorentzen J A, Ananiev E, Chamberlin M, Niu X, Meeley R et al. 2002 The defective kernel1 (dek1) gene required for aleurone cell development in the endosperm of maize grains encodes a membrane protein of the calpain gene superfamily; Proc. Natl. Acad. Sci. USA 99 5460–5465

    Article  CAS  PubMed  Google Scholar 

  • Linthorst H J M, van der Does C, Brederode F T H and Bol J F 1993 Circadian expression and induction by wounding of tobacco genes for cysteine proteinase; Plant Mol. Biol. 21 685–694

    Article  CAS  PubMed  Google Scholar 

  • Livak K J and Schmittgen T D 2001 Analysis of relative gene expression data using real-time quantitative PCR and the 2-(Delta Delta C(T)) method; Methods 25 402–408

    Article  CAS  PubMed  Google Scholar 

  • Masatoshi U, Satoshi T, Hitoshi M, Yoshimi S and Yuzo S 2007 Characterization of cysteine protease induced by oxidative stress in cells of Chlamydomonas sp. strain W80; Physiol. Plantarum 131 519–526

    Article  Google Scholar 

  • Mao X G, Zhang H Y, Tian S J, Chang X P, Jing R L 2010 TaSnRK2.4, an SNF1-type serine/threonine protein kinase of wheat (Triticum aestivum L.), confers enhanced multistress tolerance in Arabidopsis; J. Exp. Bot. 61 683–696

    Article  CAS  PubMed  Google Scholar 

  • Okamoto T and Minamikawa T 1998 A vacuolar cysteine endopeptidase (SH-EP) that digests seed storage globulin: characterization, regulation of gene expression, and posttranslational processing; J. Plant Physiol. 152 675–682

    CAS  Google Scholar 

  • Okamoto T, Shimada T, Hara N I, Nishimura M and Minamikawa T 2003 C-terminal KDEL sequence of a KDEL-tailed cysteine proteinase (sulfhydryl-endopeptidase) is involved in formation of KDEL vesicle in efficient vacuolar transport of sulfhydrylendopeptidase; Plant Physiol. 132 1892–1900

    Article  CAS  PubMed  Google Scholar 

  • Otegui M S, Noh Y S, Martinez D E, Petroff M G V, Staehlin L A and Amasino R M 2005 Senescence-associated vacuoles with intense proteolytic activity develop in leaves of Arabidopsis and soybean; Plant J. 41 831–844

    Article  CAS  PubMed  Google Scholar 

  • Pang X B, Mao X G, Jing R L, Shi J F, Gao T, Chang X P and Li Y F 2007 Analysis of gene expression profile in response to water stress in wheat (Triticum aestivum L.) seedlings; Acta Agron. Sin. 33 333–336

    CAS  Google Scholar 

  • Peng S Q, Zhu J H, Li H L and Tian W M 2008 Cloning and characterization of a novel cysteine protease gene (HbCP1) from Hevea brasiliensis; J. Biosci. 33 681–690

    Article  CAS  PubMed  Google Scholar 

  • Pyo H, Demura T and Fukuda H 2004 Spatial and temporal tracing of vessel differentiation in young Arabidopsis seedlings by the expression of an immature tracheary element-specific promoter; Plant Cell Physiol. 45 1529–1536

    Article  CAS  PubMed  Google Scholar 

  • Ramanjulu S and Bartels D 2002 Drought- and desiccation-induced modulation of gene expression in plants; Plant, Cell Environ. 25 141–151

    Article  CAS  Google Scholar 

  • Schaffer M A and Fischer R L 1988 Analysis of mRNAs that accumulate in response to low temperature indicated a thiol protease gene in tomato; Plant Physiol. 87 431–436

    Article  CAS  PubMed  Google Scholar 

  • Scharrenberg C, Quast S, Haussuhl K, Falk J and Humbeck K 2003 Isolation of senescence related cDNAs from flag leaves of field grown barley plants; Physiol. Plantarum 118 278–288

    Article  CAS  Google Scholar 

  • Sheok S, Dahiya P, Vincent J L and Brewin N J 2005 Modified expression of cysteine protease affects seed germination, vegetative growth and nodule development in transgenic lines of Medicago truncatula; Plant Sci. 169 966–975

    Article  Google Scholar 

  • Solomon M, Belenghi B, Delledonne M, Menachem E and Levine A 1999 The involvement of cysteine proteases and protease inhibitor genes in the regulation of programmed cell death in plants; Plant Cell 11 431–443

    Article  CAS  PubMed  Google Scholar 

  • Stroeher V L, Maclagan J L and Good A G 1997 Molecular cloning of a Brassica napus cysteine protease gene inducible by drought and low temperature stress; Physiol. Plantarum 101 389–397

    Article  CAS  Google Scholar 

  • Subbaiah C C, Kollipara K P and Sachs M M 2000 A Ca2+-dependent cysteine protease is associated with anoxia-induced root tip death in maize; J. Exp. Bot. 51 721–730

    Article  CAS  PubMed  Google Scholar 

  • Tabaeizadeh Z 1998 Drought-induced responses in plant cells; Int. Rev. Cytol. 182 193–247

    Article  CAS  PubMed  Google Scholar 

  • Thomashow M F 1999 Plant cold acclimation: freezing tolerance genes and regulatory mechanisms; Annu. Rev. Plant Physiol. Plant Mol. Biol. 50 571–599

    Article  CAS  PubMed  Google Scholar 

  • Ueda T, Seo S, Ohashi Y and Hashimoto J 2000 Circadian and senescence-enhanced expression of a tobacco cysteine protease gene; Plant Mol. Biol. 44 649–657

    Article  CAS  PubMed  Google Scholar 

  • Wagstaff C, Leverentz M K, Griffiths G, Thomas B and Chanasut U 2002 Cysteine protease gene expression and proteolytic activity during senescence of Alstroemeria petals; J. Exp. Bot. 367 233–240

    Article  Google Scholar 

  • Wang Z, Zang Q W, Guo Z A and Jing R L 2004 A prelinary study on gene expression profile induced by water stress in wheat (Triticum aestivum L.) seedlings; Acta Genet. Sin. 31 842–849

    CAS  PubMed  Google Scholar 

  • Williams J, Bulman M, Huttly A, Phillips A and Neill S 1994 Characterization of a cDNA from Arabidopsis thaliana encoding a potential thiol protease whose expression is induced independented by wilting and abscisic acid; Plant Mol. Biol. 25 259–270

    Article  CAS  PubMed  Google Scholar 

  • Wisniewski K and Zagdanska B 2001 Genotype-dependent proteolytic response of spring wheat to water deficiency; J. Exp. Bot. 52 1455–1463

    Article  CAS  PubMed  Google Scholar 

  • Xiong L, Schumaker K and Zhu J K 2002 Cell signaling during cold, drought, and salt stress; Plant Cell 14 S165–S183

    Article  CAS  PubMed  Google Scholar 

  • Xu C Y, Jing R L, Mao X G, Jia X Y and Chang X P 2007 A wheat (Triticum aestivum) protein phosphatase 2A catalytic subunit gene provides enhanced drought tolerance in tobacco; Ann. Bot. 93 439–450

    Article  Google Scholar 

  • Yan L F, Han J G, Yang Q C, Sun Y and Kang J M 2008 Isolation and characterization of a cDNA encoding a papain-like cysteine protease from alfalfa; DNA Sequence 19 274–281

    CAS  PubMed  Google Scholar 

  • Ye Z H and Varner J E 1996 Induction of cysteine and serine proteases during xylogenesis in Zinnia elegans; Plant Mol. Biol. 30 1233–1246

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. The National Key Facility for Crop Gene Resources and Genetic Improvement, Key Laboratory of Crop Germplasm & Biotechnology, Ministry of Agriculture, Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Qing-Wei Zang, Cai-Xiang Wang, Xu-Yan Li, Zhi-Ai Guo, Rui-Lian Jing & Xiao-Ping Chang

  2. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Jun Zhao

Authors
  1. Qing-Wei Zang
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  2. Cai-Xiang Wang
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  3. Xu-Yan Li
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  4. Zhi-Ai Guo
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  5. Rui-Lian Jing
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  6. Jun Zhao
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  7. Xiao-Ping Chang
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Correspondence to Rui-Lian Jing.

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Zang, QW., Wang, CX., Li, XY. et al. Isolation and characterization of a gene encoding a polyethylene glycol-induced cysteine protease in common wheat. J Biosci 35, 379–388 (2010). https://doi.org/10.1007/s12038-010-0043-1

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  • DOI: https://doi.org/10.1007/s12038-010-0043-1

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