Skip to main content
SpringerLink
Log in

Identification of wheat non-specific lipid transfer proteins involved in chilling tolerance

  • Original Paper
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Key message

Three TaLTPs were found to enhance chilling tolerance of transgenic Arabidopsis, which were characterized by analyzes of promoter-GUS activity, subcellular localization, chromosomal location and transcriptional profile.

Abstract

Non-specific lipid transfer proteins (nsLTP) are abundantly expressed in plants, however, their functions are still unclear. In this study, we primarily characterized the functions of 3 type I TaLTP genes that were localized on chromosomes 3A, 3B, and 5D, respectively. The transcripts of TaLTPIb.1 and TaLTPIb.5 were induced under chilling, wound, and drought conditions, while TaLTPId.1 was only up-regulated by dark treatment. All the 3 TaLTP genes could be stimulated by the in vitro treatment of salicylic acid, while TaLTPId.1 was also positively regulated by methyljasmonic acid. Furthermore, the promoter-reporter assay of TaLTPIb.1 in the transgenic brachypodium showed a typical epidermis-specific expression pattern of this gene cluster. When fused with EGFP, all the 3 proteins were shown to localize on the plasma membrane in transgenic tobacco, although a signal in chloroplasts was also observed for TaLTPId.1. Heterogeneous overexpression of each of the TaLTP genes in Arabidopsis resulted in longer root length compared with wild type plants under chilling condition. These results suggest that type I TaLTPs may have a conserved functionality in chilling tolerance by lipid permeation in the plasma membrane of epidermal cells. On the other hand, the type I TaLTPs may exert functional divergence mainly through regulatory subfunctionalization.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Amid A, Lytovchenko A, Fernie AR, Warren G, Thorlby GJ (2012) The sensitive to freezing3 mutation of Arabidopsis thaliana is a cold-sensitive allele of homomeric acetyl-CoA carboxylase that results in cold-induced cuticle deficiencies. J Exp Bot 63:5289–5299

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Blein JP, Coutos-Thevenot P, Marion D, Ponchet M (2002) From elicitins to lipid-transfer proteins: a new insight in cell signalling involved in plant defence mechanisms. Trends Plant Sci 7:293–296

  • Boutrot F, Chantret N, Gautier MF (2008) Genome-wide analysis of the rice and Arabidopsis non-specific lipid transfer protein (nsLtp) gene families and identification of wheat nsLtp genes by EST data mining. BMC Genom 9:86

    Article  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

  • Bragg JN, Wu J, Gordon SP, Guttman ME, Thilmony R, Lazo GR, Gu YQ, Vogel JP (2012) Generation and characterization of the Western Regional Research Center Brachypodium T-DNA insertional mutant collection. PLoS One 7:e41916

  • Brenchley R, Spannagl M, Pfeifer M, Barker GL, D’Amore R, Allen AM, McKenzie N, Kramer M, Kerhornou A, Bolser D, Kay S, Waite D, Trick M, Bancroft I, Gu Y, Huo N, Luo MC, Sehgal S, Gill B, Kianian S, Anderson O, Kersey P, Dvorak J, McCombie WR, Hall A, Mayer KF, Edwards KJ, Bevan MW, Hall N (2012) Analysis of the bread wheat genome using whole-genome shotgun sequencing. Nature 491:705–710

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Caaveiro JM, Molina A, Gonzalez-Manas JM, Rodriguez-Palenzuela P, Garcia-Olmedo F, Goni FM (1997) Differential effects of five types of antipathogenic plant peptides on model membranes. FEBS Lett 410:338–342

  • Carvalho Ade O, Gomes VM (2007) Role of plant lipid transfer proteins in plant cell physiology—a concise review. Peptides 28:1144–1153

  • Chinnusamy V, Ohta M, Kanrar S, Lee BH, Hong X, Agarwal M, Zhu JK (2003) ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes Dev 17:1043–1054

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743

    Article  PubMed  CAS  Google Scholar 

  • Das P, Ito T, Wellmer F, Vernoux T, Dedieu A, Traas J, Meyerowitz EM (2009) Floral stem cell termination involves the direct regulation of AGAMOUS by PERIANTHIA. Development 136:1605–1611

    Article  PubMed  CAS  Google Scholar 

  • George S, Parida A (2010) Characterization of an oxidative stress inducible nonspecific lipid transfer protein coding cDNA and its promoter from drought tolerant plant prosopis juliflora. Plant Mol Biol Rep 28:32–40

  • Guo L, Yang H, Zhang X, Yang S (2013) Lipid transfer protein 3 as a target of MYB96 mediates freezing and drought stress in Arabidopsis. J Exp Bot 64:1755–1767

  • Han GW, Lee JY, Song HK, Chang C, Min K, Moon J, Shin DH, Kopka ML, Sawaya MR, Yuan HS, Kim TD, Choe J, Lim D, Moon HJ, Suh SW (2001) Structural basis of non-specific lipid binding in maize lipid-transfer protein complexes revealed by high-resolution X-ray crystallography. J Mol Biol 308:263–278

  • Huang MD, Chen TL, Huang AH (2013) Abundant type III lipid transfer proteins in Arabidopsis tapetum are secreted to the locule and become a constituent of the pollen exine. Plant Physiol 163:1218–1229

  • Hull GA, Devic M (1995) The beta-glucuronidase (gus) reporter gene system. Gene fusions; spectrophotometric, fluorometric, and histochemical detection. Methods Mol Biol 49:125–141

    PubMed  CAS  Google Scholar 

  • Jang CS, Yim WC, Moon JC, Hung JH, Lee TG, Lim SD, Cho SH, Lee KK, Kim W, Seo YW, Lee BM (2008) Evolution of non-specific lipid transfer protein (nsLTP) genes in the Poaceae family: their duplication and diversity. Mol Genet Genomics 279:481–497

  • Jung HW, Kim KD, Hwang BK (2005) Identification of pathogen-responsive regions in the promoter of a pepper lipid transfer protein gene (CALTPI) and the enhanced resistance of the CALTPI transgenic Arabidopsis against pathogen and environmental stresses. Planta 221:361–373

  • Kader JC (1996) Lipid-transfer proteins in plants. Annu Rev Plant Physiol Plant Mol Biol 47:627–654

    Article  PubMed  CAS  Google Scholar 

  • Kristensen AK, Brunstedt J, Nielsen KK, Roepstorff P, Mikkelsen JD (2000) Characterization of a new antifungal non-specific lipid transfer protein (nsLTP) from sugar beet leaves. Plant Sci 155:31–40

    Article  PubMed  CAS  Google Scholar 

  • Kumar S, Nei M, Dudley J, Tamura K (2008) MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Brief Bioinform 9:299–306

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Lai WC, Wang HC, Chen GY, Yang JC, Korinek M, Hsieh CJ, Nozaki H, Hayashi K, Wu CC, Wu YC, Chang FR (2011) Using the pER8:GUS reporter system to screen for phytoestrogens from Caesalpinia sappan. J Nat Prod 74:1698–1706

    Article  PubMed  CAS  Google Scholar 

  • Lee JY, Min K, Cha H, Shin DH, Hwang KY, Suh SW (1998) Rice non-specific lipid transfer protein: the 1.6 A crystal structure in the unliganded state reveals a small hydrophobic cavity. J Mol Biol 276:437–448

  • Lee SB, Go YS, Bae HJ, Park JH, Cho SH, Cho HJ, Lee DS, Park OK, Hwang I, Suh MC (2009) Disruption of glycosylphosphatidylinositol-anchored lipid transfer protein gene altered cuticular lipid composition, increased plastoglobules, and enhanced susceptibility to infection by the fungal pathogen Alternaria brassicicola. Plant Physiol 150:42–54

  • Lei L, Chen L, Shi X, Li Y, Wang J, Chen D, Xie F (2014) A nodule-specific lipid transfer protein AsE246 participates in transport of plant-synthesized lipids to symbiosome membrane and is essential for nodule organogenesis in Chinese milk vetch. Plant Physiol 164:1045–1058

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Maldonado AM, Doerner P, Dixon RA, Lamb CJ, Cameron RK (2002) A putative lipid transfer protein involved in systemic resistance signalling in Arabidopsis. Nature 419:399–403

  • Mollet JC, Park SY, Nothnagel EA, Lord EM (2000) A lily stylar pectin is necessary for pollen tube adhesion to an in vitro stylar matrix. Plant Cell 12:1737–1750

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Nelson BK, Cai X, Nebenfuhr A (2007) A multicolored set of in vivo organelle markers for co-localization studies in Arabidopsis and other plants. Plant J 51:1126–1136

    Article  PubMed  CAS  Google Scholar 

  • Paterson AH, Bowers JE, Bruggmann R, Dubchak I, Grimwood J, Gundlach H, Haberer G, Hellsten U, Mitros T, Poliakov A, Schmutz J, Spannagl M, Tang H, Wang X, Wicker T, Bharti AK, Chapman J, Feltus FA, Gowik U, Grigoriev IV, Lyons E, Maher CA, Martis M, Narechania A, Otillar RP, Penning BW, Salamov AA, Wang Y, Zhang L, Carpita NC, Freeling M, Gingle AR, Hash CT, Keller B, Klein P, Kresovich S, McCann MC, Ming R, Peterson DG, Mehboobur R, Ware D, Westhoff P, Mayer KF, Messing J, Rokhsar DS (2009) The Sorghum bicolor genome and the diversification of grasses. Nature 457:551–556

    Article  PubMed  CAS  Google Scholar 

  • Pitzschke A, Datta S, Persak H (2013) Salt stress in Arabidopsis: lipid transfer protein AZI1 and its control by Mitogen-activated protein kinase MPK3. Mol Plant

  • Pyee J, Yu H, Kolattukudy PE (1994) Identification of a lipid transfer protein as the major protein in the surface wax of broccoli (Brassica oleracea) leaves. Arch Biochem Biophys 311:460–468

    Article  PubMed  CAS  Google Scholar 

  • Samuel D, Liu YJ, Cheng CS, Lyu PC (2002) Solution structure of plant nonspecific lipid transfer protein-2 from rice (Oryza sativa). J Biol Chem 277:35267–35273

  • Sanghera GS, Wani SH, Hussain W, Singh NB (2011) Engineering cold stress tolerance in crop plants. Curr Genomics 12:30–43

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  • Shin DH, Lee JY, Hwang KY, Kim KK, Suh SW (1995) High-resolution crystal structure of the non-specific lipid-transfer protein from maize seedlings. Structure 3:189–199

    Article  PubMed  CAS  Google Scholar 

  • Thoma S, Hecht U, Kippers A, Botella J, De Vries S, Somerville C (1994) Tissue-specific expression of a gene encoding a cell wall-localized lipid transfer protein from Arabidopsis. Plant Physiol 105:35–45

  • Tian A, Jiang J, Cao J (2013) Functional analysis of a novel male fertility lipid transfer protein gene in Brassica campestris ssp. chinensis. Plant Mol Biol Rep 31:775–782

  • Tsuboi S, Osafune T, Tsugeki R, Nishimura M, Yamada M (1992) Nonspecific lipid transfer protein in castor bean cotyledon cells: subcellular localization and a possible role in lipid metabolism. J Biochem 111:500–508

    PubMed  CAS  Google Scholar 

  • Vergnolle C, Arondel V, Jolliot A, Kader JC (1992) Phospholipid transfer proteins from higher plants. Methods Enzymol 209:522–530

    Article  PubMed  CAS  Google Scholar 

  • Wang HW, Kwon HJ, Yim WC, Lim SD, Moon JC, Lee BM, Seo YW, Kim W, Jang CS (2010) Expressional diversity of wheat nsLTP genes: evidence of subfunctionalization via cis-regulatory divergence. Genetica 138:843–852

  • Wang HW, Hwang SG, Karuppanapandian T, Liu A, Kim W, Jang CS (2012) Insight into the molecular evolution of non-specific lipid transfer proteins via comparative analysis between rice and sorghum. DNA Res 19:179–194

    Article  PubMed  PubMed Central  Google Scholar 

  • Zachowski A, Guerbette F, Grosbois M, Jolliot-Croquin A, Kader JC (1998) Characterisation of acyl binding by a plant lipid-transfer protein. Eur J Biochem 257:443–448

Download references

Acknowledgments

This work was financially supported by National Natural Science Foundation of China (No. 31200982), 973 Developmental Program (2014CB138100), and Shandong province programs (20133702120002 and BS2013NY006).

Conflict of interest

We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled.

Author information

Authors and Affiliations

  1. State Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, 61 Daizong Street, Tai’an, 271018, Shandong, People’s Republic of China

    Guanghui Yu, Wenqian Hou, Xuye Du, Liang Wang, Hongyan Wu, Lanfei Zhao, Lingrang Kong & Hongwei Wang

Authors
  1. Guanghui Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wenqian Hou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xuye Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Liang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hongyan Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Lanfei Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Lingrang Kong
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hongwei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Lingrang Kong or Hongwei Wang.

Additional information

Communicated by Baochun Li.

G. Yu and W. Hou contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOC 46 kb)

Supplementary material 2 (PPTX 3715 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yu, G., Hou, W., Du, X. et al. Identification of wheat non-specific lipid transfer proteins involved in chilling tolerance. Plant Cell Rep 33, 1757–1766 (2014). https://doi.org/10.1007/s00299-014-1655-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-014-1655-y

Keywords

Search

Navigation