Register      Login
Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Promoter of the wheat lipid transfer protein, TdLTP4, drives leaf-preferential expression in transgenic Arabidopsis plants

Héla Safi A , Nebras Belgaroui A , Khaled Masmoudi A B and Faiçal Brini https://orcid.org/0000-0002-8435-381X A C
+ Author Affiliations
- Author Affiliations

A Biotechnology and Plant Improvement Laboratory, Centre of Biotechnology of Sfax (CBS)/University of Sfax, BP ‘1177’ 3018, Sfax – Tunisia.

B Present address: College of Food and Agriculture, Arid land department, United Arab Emirates University, UAE.

C Corresponding author. Email: faical.brini@cbs.rnrt.tn

Functional Plant Biology 46(3) 275-285 https://doi.org/10.1071/FP18040
Submitted: 16 February 2018  Accepted: 15 October 2018   Published: 1 November 2018

Abstract

In a previous report, a gene encoding a durum wheat lipid transfer protein, TdLTP4, was characterised as induced by abiotic and biotic stresses. In the present work, we investigated the regulation of the gene TdLTP4. A TdLTP4 promoter (PrTdLTP4) region of around 868-bp was isolated and sequenced. Its analysis revealed the presence of several DNA boxes known to be important mainly in the regulation of genes expressed under abiotic stress (salt and dehydration), abscisic acid (ABA) and pathogen responsiveness. The whole PrTdLTP4 fragment was fused to the reporter gene β-glucuronidase (gusA) and analysed in transgenic Arabidopsis plants. Histochemical assays of transgenic Arabidopsis plants showed that the 868-bp fragment of TdLTP4 gene promoter was found to be sufficient for both spatial and temporal patterns of its expression. Under control conditions, GUS histochemical staining was observed significantly only in young leaves of 8- and 12-day-old plants. Whereas after stress challenge especially with NaCl and mannitol, GUS transcripts expression increased substantially in leaves of 30-day-old transgenic seedlings. Real-time qPCR expression analysis of the gusA gene, confirmed the results of histochemical assays. Taken together these data provide evidence that PrTdLTP4 functions as abiotic-stress-inducible promoter in a heterologous dicot system and could be an excellent tool for future crop improvement.

Additional keywords: abiotic stress, GUS activity, PrTdLTP4 promoter, transgenic Arabidopsis, Triticum durum.


References

Ben Saad R, Fabre D, Mieulet D, Meynard D, Dingkuhn M, Al-Doss A, Guiderdoni E, Hassairi A (2012) Expression of the Aeluropus littoralis AlSAP gene in rice confers broad tolerance to abiotic stresses through maintenance of photosynthesis. Plant, Cell & Environment 35, 626–643.
Expression of the Aeluropus littoralis AlSAP gene in rice confers broad tolerance to abiotic stresses through maintenance of photosynthesis.Crossref | GoogleScholarGoogle 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 in Plant Science 7, 293–296.
From elicitins to lipid-transfer proteins: a new insight in cell signalling involved in plant defence mechanisms.Crossref | GoogleScholarGoogle Scholar |

Canevascini S, Caderas D, Mandel T, Fleming AJ, Dupuis I, Kuhlemeier C (1996) Tissue-specific expression and promoter analysis of the tobacco ltp1 gene. Plant Physiology 112, 513–524.
Tissue-specific expression and promoter analysis of the tobacco ltp1 gene.Crossref | GoogleScholarGoogle Scholar |

Carvalho AO, Gomes VM (2007) Role of plant lipid transfer proteins in plant cell physiology: a concise review. Peptides 28, 1144–1153.
Role of plant lipid transfer proteins in plant cell physiology: a concise review.Crossref | GoogleScholarGoogle Scholar |

Chen H, Nelson RS, Sherwood JL (1994) Enhanced recovery of transformants of Agrobacterium tumefaciens after freeze-thaw transformation and drug selection. BioTechniques 16, 664–668, 670.

Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. The Plant Journal 16, 735–743.
Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.Crossref | GoogleScholarGoogle Scholar |

Fleming AJ, Mandel T, Hofmann S, Sterk P, de Vries SC, Kuhlemeier C (1992) Expression pattern of a tobacco lipid transfer protein gene within the shoot apex. The Plant Journal 2, 855–862.

Gomès E, Sagot E, Gaillard C, Laquitaine L, Poinssot B, Sanejouand Y, Delrop S, Coutos-Thévenot P (2003) Nonspecific lipid transfer protein genes expression in grape (Vitis sp.) cells in response to fungal elicitor treatments. Molecular Plant-Microbe Interactions 16, 456–464.
Nonspecific lipid transfer protein genes expression in grape (Vitis sp.) cells in response to fungal elicitor treatments.Crossref | GoogleScholarGoogle Scholar |

Guiderdoni E, Cordero MJ, Vignols F, Garcia-Garrido JM, Lescot M, Tharreau D, Meynard D, Ferrie’re N, Notteghem J, Deldeny M (2002) Inducibility by pathogen attack and developmental regulation of the rice Ltp1 gene. Plant Molecular Biology 49, 679–695.
Inducibility by pathogen attack and developmental regulation of the rice Ltp1 gene.Crossref | GoogleScholarGoogle Scholar |

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. Journal of Molecular Biology 308, 263–278.
Structural basis of non-specific lipid binding in maize lipid-transfer protein complexes revealed by high-resolution x-ray crystallography.Crossref | GoogleScholarGoogle Scholar |

Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database: 1999. Nucleic Acids Research 27, 297–300.
Plant cis-acting regulatory DNA elements (PLACE) database: 1999.Crossref | GoogleScholarGoogle Scholar |

Hsieh TH, Lee JT, Charng YY, Chan MT (2002) Tomato plants ectopically expressing Arabidopsis CBF1 show enhanced resistance to water deficit stress. Plant Physiology 130, 618–626.
Tomato plants ectopically expressing Arabidopsis CBF1 show enhanced resistance to water deficit stress.Crossref | GoogleScholarGoogle Scholar |

Hughes MA, Dunn MA, Pearce RS, White AJ, Zhang L (1992) An abscisic acid-responsive, low temperature barley gene has homology with a maize phospholipid transfer protein. Plant, Cell & Environment 15, 861–865.
An abscisic acid-responsive, low temperature barley gene has homology with a maize phospholipid transfer protein.Crossref | GoogleScholarGoogle Scholar |

Ito Y, Katsura K, Maruyama K, Taji T, Kobayashi M, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2006) Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice. Plant & Cell Physiology 47, 141–153.
Functional analysis of rice DREB1/CBF-type transcription factors involved in cold-responsive gene expression in transgenic rice.Crossref | GoogleScholarGoogle Scholar |

Iwamoto M, Higo H, Higo K (2004) Strong expression of the rice catalase gene CatB promoter in protoplasts and roots of both monocots and dicots. Plant Physiology and Biochemistry 42, 241–249.
Strong expression of the rice catalase gene CatB promoter in protoplasts and roots of both monocots and dicots.Crossref | GoogleScholarGoogle Scholar |

Jakobsen K, Klemsdal SS, Aalen RB, Bosnes M, Alexander D, Olsen OA (1989) Barley aleurone development: molecular cloning of aleurone-specific cDNAs from immature grains. Plant Molecular Biology 12, 285–293.
Barley aleurone development: molecular cloning of aleurone-specific cDNAs from immature grains.Crossref | GoogleScholarGoogle Scholar |

Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. The EMBO Journal 6, 3901–3907.
GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.Crossref | GoogleScholarGoogle Scholar |

Jung HW, Kim W, Hwang BK (2003) Three pathogen-inducible genes encoding lipid transfer protein from pepper are differentially activated by pathogens, abiotic, and environmental stresses. Plant, Cell & Environment 26, 915–928.
Three pathogen-inducible genes encoding lipid transfer protein from pepper are differentially activated by pathogens, abiotic, and environmental stresses.Crossref | GoogleScholarGoogle Scholar |

Kader JC (1996) Lipid transfer proteins in plants. Annual Review of Plant Physiology and Plant Molecular Biology 47, 627–654.
Lipid transfer proteins in plants.Crossref | GoogleScholarGoogle Scholar |

Kanneganti V, Gupta AK (2008) Overexpression of OsiSAP8, a member of stress associated protein (SAP) gene family of rice confers tolerance to salt, drought and cold stress in transgenic tobacco and rice. Plant Molecular Biology 66, 445–462.
Overexpression of OsiSAP8, a member of stress associated protein (SAP) gene family of rice confers tolerance to salt, drought and cold stress in transgenic tobacco and rice.Crossref | GoogleScholarGoogle Scholar |

Kim TH, Park JH, Kim MC, Cho SH (2008) Cutin monomer induces expression of the rice OsLTP5 lipid transfer protein gene. Journal of Plant Physiology 165, 345–349.
Cutin monomer induces expression of the rice OsLTP5 lipid transfer protein gene.Crossref | GoogleScholarGoogle Scholar |

Kurek I, Stoger E, Dulberger R, Christou P, Breiman A (2002) Overexpression of the wheat FK506-binding protein 73 (FKBP73) and the heat-induced wheat FKBP77 in transgenic wheat reveals different functions of the two isoforms. Transgenic Research 11, 373–379.
Overexpression of the wheat FK506-binding protein 73 (FKBP73) and the heat-induced wheat FKBP77 in transgenic wheat reveals different functions of the two isoforms.Crossref | GoogleScholarGoogle Scholar |

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

Lemieux B (1996) Molecular genetics of epicuticular wax biosynthesis. Trends in Plant Science 1, 312–318.
Molecular genetics of epicuticular wax biosynthesis.Crossref | GoogleScholarGoogle Scholar |

Lescot M, Déhais P, Thijs G, Marchal K, Moreau Y, Van de Peer Y, Rouzé P, Rombauts S (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Research 30, 325–327.
PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences.Crossref | GoogleScholarGoogle Scholar |

Linnestad C, Lönneborg A, Kalla R, Olsen OA (1991) Promoter of a lipid transfer protein gene expressed in barley aleurone cells contains similar myb and myc recognition sites as the maize Bz-McC allele. Plant Physiology 97, 841–843.
Promoter of a lipid transfer protein gene expressed in barley aleurone cells contains similar myb and myc recognition sites as the maize Bz-McC allele.Crossref | GoogleScholarGoogle Scholar |

Liu ZZ, Wang JL, Huang X, Xu WH, Liu ZM, Fang RX (2003) The promoter of a rice glycine-rich protein gene, Osgrp-2, confers vascular-specific expression in transgenic plants. Planta 216, 824–833.

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔC T method. Methods 25, 402–408.
Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔC T method.Crossref | GoogleScholarGoogle Scholar |

Maldonado AM, Doerner P, Dixon RA, Lamb CJ, Cameron RK (2002) A putative lipid transfer protein involved in systemic resistance signaling in Arabidopsis. Nature 419, 399–403.
A putative lipid transfer protein involved in systemic resistance signaling in Arabidopsis.Crossref | GoogleScholarGoogle Scholar |

Maruyama K, Todaka D, Mizoi J, Yoshida T, Kidokoro S, Matsukura S, Takasaki H, Sakurai T, Yamamoto YY, Yoshiwara K, Kojima M, Sakakibara H, Shinozaki K, Yamaguchi-Shinozaki K (2012) Identification of cis-acting promoter elements in cold- and dehydration-induced transcriptional pathways in Arabidopsis, rice, and soybean. DNA Research 19, 37–49.
Identification of cis-acting promoter elements in cold- and dehydration-induced transcriptional pathways in Arabidopsis, rice, and soybean.Crossref | GoogleScholarGoogle Scholar |

Michiels A, Tucker M, Van den Ende W, Van Laere A (2003a) Chromosomal walking of flanking regions from short known sequences in GC-rich plant genomic DNA. Plant Molecular Biology Reporter 21, 295–302.
Chromosomal walking of flanking regions from short known sequences in GC-rich plant genomic DNA.Crossref | GoogleScholarGoogle Scholar |

Michiels A, Van den Ende W, Tucker M, Van Riet L, Van Laere A (2003b) Extraction of high-quality genomic DNA from latex-containing plants. Analytical Biochemistry 315, 85–89.
Extraction of high-quality genomic DNA from latex-containing plants.Crossref | GoogleScholarGoogle Scholar |

Molina A, Garcia-Olmedo F (1993) Developmental and pathogen induced expression of three barley genes encoding lipid transfer protein. The Plant Journal 4, 983–991.
Developmental and pathogen induced expression of three barley genes encoding lipid transfer protein.Crossref | GoogleScholarGoogle Scholar |

Nielsen KK, Nielsen JE, Madrid SM, Mikkelsen JD (1996) New antifungal proteins from sugar beet (Beta vulgaris L.) showing homology to non-specific lipid transfer proteins. Plant Molecular Biology 31, 539–552.
New antifungal proteins from sugar beet (Beta vulgaris L.) showing homology to non-specific lipid transfer proteins.Crossref | GoogleScholarGoogle Scholar |

Romero C, Bellés J, Vayá J, Serrano R, Culiáñez-Macià F (1997) Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance. Planta 201, 293–297.
Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiotropic phenotypes include drought tolerance.Crossref | GoogleScholarGoogle Scholar |

Rozen S, Skaletsky HJ (2000) Primer3 on the WWW for general users and for biologist programmers. Methods in Molecular Biology (Clifton, N.J.) 132, 365–386.

Safi H, Saibi W, Alaoui MM, Hmyene A, Masmoudi K, Hanin M, Brini F (2015) A wheat lipid transfer protein (TdLTP4) promotes tolerance to abiotic and biotic stress in Arabidopsis thaliana. Plant Physiology and Biochemistry 89, 64–75.
A wheat lipid transfer protein (TdLTP4) promotes tolerance to abiotic and biotic stress in Arabidopsis thaliana.Crossref | GoogleScholarGoogle Scholar |

Salminen TA, Eklund DM, Joly V, Blomqvist K, Matton DP, Edqvist J (2018) Deciphering the evolution and development of the cuticle by studying lipid transfer proteins in mosses and liverworts. Plants 7, 6
Deciphering the evolution and development of the cuticle by studying lipid transfer proteins in mosses and liverworts.Crossref | GoogleScholarGoogle Scholar |

Samuel D, Liu YJ, Cheng CS, Lyu PC (2002) Solution structure of plant nonspecific lipid transfer protein-2 from rice (Oryza sativa). Journal of Biological Chemistry 277, 35267–35273.
Solution structure of plant nonspecific lipid transfer protein-2 from rice (Oryza sativa).Crossref | GoogleScholarGoogle Scholar |

Sohal AK, Pallas JA, Jenkins GI (1999) The promoter of a Brassica napus lipid transfer protein gene is active in a range of tissues and stimulated by light and viral infection in transgenic Arabidopsis. Plant Molecular Biology 41, 75–87.
The promoter of a Brassica napus lipid transfer protein gene is active in a range of tissues and stimulated by light and viral infection in transgenic Arabidopsis.Crossref | GoogleScholarGoogle Scholar |

Sterk P, Booij H, Schellekens GA, Van Kammen A, De Vries SC (1991) Cell-specific expression of the carrot EP2 lipid transfer protein gene. The Plant Cell 3, 907–921.
Cell-specific expression of the carrot EP2 lipid transfer protein gene.Crossref | GoogleScholarGoogle Scholar |

Tapia G, Morales-Quintana L, Parra C, Berbel A, Alcorta M (2013) Study of nsLTPs in Lotus japonicus genome reveal a specific epidermal cell member (LjLTP10) regulated by drought stress in aerial organs with a putative role in cutin formation. Plant Molecular Biology 82, 485–501.
Study of nsLTPs in Lotus japonicus genome reveal a specific epidermal cell member (LjLTP10) regulated by drought stress in aerial organs with a putative role in cutin formation.Crossref | GoogleScholarGoogle Scholar |

Thoma SL, Kaneko Y, Somerville C (1993) An Arabidopsis lipid transfer protein is a cell wall protein. The Plant Journal 3, 427–436.
An Arabidopsis lipid transfer protein is a cell wall protein.Crossref | GoogleScholarGoogle Scholar |

Thoma S, Hecht U, Kippers A, Botella J, De Vries S, Somerville CR (1994) Tissue-specific expression of a gene encoding a cell wall-localized lipid transfer protein from Arabidopsis. Plant Physiology 105, 35–45.
Tissue-specific expression of a gene encoding a cell wall-localized lipid transfer protein from Arabidopsis.Crossref | GoogleScholarGoogle Scholar |

Tittarelli A, Milla L, Vargas F, Morales A, Neupert C, Meisel L, Salvo-G H, Penaloza L, Munoz E, Corcuela E, Silva H (2007) Isolation and comparative analysis of the wheat TaPT2 promoter: identification in silico of new putative regulatory motifs conserved between monocots and dicots. Journal of Experimental Botany 58, 2573–2582.
Isolation and comparative analysis of the wheat TaPT2 promoter: identification in silico of new putative regulatory motifs conserved between monocots and dicots.Crossref | GoogleScholarGoogle Scholar |

Torres-Schumann S, Godoy JA, Pintor-Toro JA (1992) A probable lipid transfer protein gene is induced by NaCl in stems of tomato plants. Plant Molecular Biology 18, 749–757.
A probable lipid transfer protein gene is induced by NaCl in stems of tomato plants.Crossref | GoogleScholarGoogle Scholar |

Tounsi S, Feki K, Saïdi MN, Maghrebi S, Brini F, Masmoudi Kh (2018) Promoter of the TmHKT1;4-A1 gene of Triticum monococcum directs stress inducible, developmental regulated and organ specific gene expression in transgenic Arabidopsis thaliana. World Journal of Microbiology & Biotechnology 34, 99
Promoter of the TmHKT1;4-A1 gene of Triticum monococcum directs stress inducible, developmental regulated and organ specific gene expression in transgenic Arabidopsis thaliana.Crossref | GoogleScholarGoogle Scholar |

Treviño MB, O’Connell MA (1998) Three drought-responsive members of the nonspecific lipid-transfer protein gene family in Lycopersicon pennellii show different developmental patterns of expression. Plant Physiology 116, 1461–1468.
Three drought-responsive members of the nonspecific lipid-transfer protein gene family in Lycopersicon pennellii show different developmental patterns of expression.Crossref | GoogleScholarGoogle Scholar |

Wang C, Xie W, Chi F, Hu W, Mao G, Sun D, Li C, Sun Y (2007) BcLTP, a novel lipid transfer protein in Brassica chinensis, may secrete and combine extracellular CaM. Plant Cell Reports 27, 159–169.
BcLTP, a novel lipid transfer protein in Brassica chinensis, may secrete and combine extracellular CaM.Crossref | GoogleScholarGoogle Scholar |

Xu R, Zhao H, Dinkins RD, Cheng X, Carberry G, Li QQ (2006) The 73 kD subunit of the cleavage and polyadenylation specificity factor (CPSF) complex affects reproductive development in Arabidopsis. Plant Molecular Biology 61, 799–815.
The 73 kD subunit of the cleavage and polyadenylation specificity factor (CPSF) complex affects reproductive development in Arabidopsis.Crossref | GoogleScholarGoogle Scholar |

Yeats TH, Rose JKC (2008) The biochemistry and biology of extracellular plant lipid-transfer proteins (LTPs). Protein Science 17, 191–198.
The biochemistry and biology of extracellular plant lipid-transfer proteins (LTPs).Crossref | GoogleScholarGoogle Scholar |

Yubero-Serrano EM, Moyano E, Medina-Escobar N, Muñoz-Blanco J, Caballero JL (2003) Identification of a strawberry gene encoding anon-specific lipid transfer protein that responds to ABA, wounding and cold stress. Journal of Experimental Botany 54, 1865–1877.
Identification of a strawberry gene encoding anon-specific lipid transfer protein that responds to ABA, wounding and cold stress.Crossref | GoogleScholarGoogle Scholar |