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

G. Tapia1, L. Morales-Quintana2, C. Parra2, A. Berbel3, M. Alcorta1
1Unidad de Recursos Genéticos, Instituto de Investigaciones Agropecuarias, INIA-Quilamapu, Chillán, Chile
2Instituto de Biología Vegetal y Biotecnología, Universidad de Talca, Talca, Chile
3Instituto de Biología Molecular y Celular de Plantas, Valencia, Spain

Tóm tắt

The cuticle is the first defense against pathogens and the second way water is lost in plants. Hydrophobic layers covering aerial plant organs from primary stages of development form cuticle, including major classes of aliphatic wax components and cutin. Extensive research has been conducted to understand cuticle formation mechanisms in plants. However, many questions remain unresolved in the transport of lipid components to form cuticle. Database studies of the Lotus japonicus genome have revealed the presence of 24 sequences classified as putative non-specific lipid transfer proteins (nsLTPs), which were classified in seven groups; four groups were selected because of their expression in aerial organs. LjLTP8 forms a cluster with DIR1 in Arabidopsis thaliana while LjLTP6, LjLTP9, and LjLTP10 were grouped as type I LTPs. In silico studies showed a high level of structural conservation, and substrate affinity studies revealed palmitoyl-CoA as the most likely ligand for these LTPs, although the Lyso-Myristoyl Phosphatidyl Choline, Lyso-myristoyl phosphatidyl glycerol, and Lyso-stearyl phosphatidyl choline ligands also showed a high affinity with the proteins. The LjLTP6 and LjLTP10 genes were expressed in both the stems and the leaves under normal conditions and were highly induced during drought stress. LjLTP10 was the most induced gene in shoots during drought. The gene was only expressed in the epidermal cells of stems, primordial leaves, and young leaflets. LjLTP10 was positively regulated by MeJA but repressed by abscisic acid (ABA), ethylene, and H2O2, while LjLTP6 was weakly induced by MeJA, repressed by H2O2, and not affected by ABA and ethylene. We suggest that LjLTP10 is involved in plant development of stem and leaf cuticle, but also in acclimation to tolerate drought stress in L. japonicus.

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Tài liệu tham khảo

Abagyan RA, Totrov MM, Kuznetsov DA (1994) Icm: a new method for protein modeling and design: applications to docking and structure prediction from the distorted native conformation. J Comput Chem 15:488–506

Acuña H, Inostroza L, Sanchez M, Tapia G (2010) Drought-tolerant naturalized populations of Lotus tenuis for constrained environments. Acta Agric Scand Sect B Soil Plant Sci 60:174–181

Acuña H, Ortega F, Seguel I, Barrientos L (2012) Introduction, collection and characterization of Lotus spp. germplasm in Chile. J Environ Sci Eng 1:533–552

Ahn S, Kim J, Pyee J, Park H (2009) Biochemical characterization of the lipid-binding properties of a broccoli cuticular wax-associated protein, WAX9D, and its application. BMB Rep 42:367–372

Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

Betti M, Pérez-Delgado C, García-Calderón M, Díaz P, Monza J, Márquez AJ (2012) Cellular stress following water deprivation in the model legume Lotus japonicus. Cells 1:1089–1106

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 Genomics 9:86

Bringe K, Schumacher CFA, Schmitz-Eiberger MA, Steiner U, Oerke EC (2006) Ontogenetic variation in chemical and physical characteristics of adaxial apple leaf surfaces. Phytochemistry 67:161–170

Broun P, Poindexter P, Osborne E, Jiang CZ, Riechmann JL (2004) WIN1, a transcriptional activator of epidermal wax accumulation in Arabidopsis. Proc Natl Acad Sci USA 101:4706–4711

Cameron KD, Teece MA, Smart LB (2006) Increased accumulation of cuticular wax and expression of lipid transfer protein in response to periodic drying events in leaves of tree tobacco. Plant Physiol 140:176–183

Chae K, Kieslich CA, Morikis D, Kim SC, Lord EM (2009) A gain-of-function mutation of arabidopsis lipid transfer protein 5 disturbs pollen tube tip growth and fertilization. Plant Cell 21:3902–3914

Chae K, Gonong BJ, Kim SC, Kieslich CA, Morikis D, Balasubramanian S, Lord EM (2010) A multifaceted study of stigma/style cysteine-rich adhesin (SCA)-like Arabidopsis lipid transfer proteins (LTPs) suggests diversified roles for these LTPs in plant growth and reproduction. J Exp Bot 61:4277–4290

Champigny MJ, Shearer H, Mohammad A, Haines K, Neumann M, Thilmony R, He S-H, Fobert P, Dengler N, Cameron RK (2011) Localization of DIR1 at the tissue, cellular and subcellular levels during systemic acquired resistance in Arabidopsis using DIR1:GUS and DIR1:EGFP reporters. BMC Plant Biol 11:125

Chanda B, Xia Y, Mandal MK, Yu K, Sekine KT, Gao QM, Selote D, Hu Y, Stromberg A, Navarre DA, Kachroo A, Kachroo P (2011) Glycerol-3-phosphate is a critical mobile inducer of systemic immunity in plants. Nat Genet 43:421–427. doi:10.1038/ng.798

Charvolin D, Douliez J-P, Marion D, Cohen-Addad C, Pebay-Pey-roula E (1999) The crystal structure of a wheat non specific lipid transfer protein (ns–LTP1) complexed with two molecules of phospholipid at 2.1 Å resolution. Eur J Biochem 264:562–568

Cheng HC, Cheng PT, Peng P, Lyu PC, Sun YJ (2004) Lipid binding in rice nonspecific lipid transfer protein-1 complexes from Oryza sativa. Protein Sci 13:2304–2315

Czechowski T, Stitt M, Altmann T, Udvardi MK, Scheible WR (2005) Genome-wide identification and testing of superior reference genes for transcript normalization in Arabidopsis. Plant Physiol 139:5–17

Da Silva VA, Landon C, Industri B, Marais A, Marion D, Ponchet M, Vovelle M (2005) Solution structure of a tobacco lipid transfer protein exhibiting new biophysical and biological features. Proteins 59:356–367

Diaz P, Borsani O, Márquez AJ, Monza J (2005) Osmotically induced proline accumulation in Lotus corniculatus leaves is affected by light and nitrogen source. Plant Growth Regul 46:223–232

Douliez JP, Michon T, Elmorjani K, Marion D (2000) Structure, biological and technological functions of lipid transfer proteins and indolines, the major lipid binding proteins from cereal kernels. J Cereal Sci 32:1–20

Duan H, Schuler MA (2005) Differential expression and evolution of the Arabidopsis CYP86A subfamily. Plant Physiol 137:1067–1081

Edstam MM, Viitanen L, Salminen TA, Edqvist J (2011) Evolutionary history of the non-specific lipid transfer proteins. Mol Plant 4:947–964

Edwards D, Abbott GD, Raven JA (1996) Cuticles of early land plants: a palaeoecophysiological evaluation. In: Kerstiens G (ed) Plant cuticles: an integrated functional approach. Bios Scientific Publishers, Oxford, pp 1–32

Fauth M, Schweizer P, Buchala A, Markstädter C, Riederer M, Kato T, Kauss H (1998) Cutin monomers and surface wax constituents elicit H2O2 in conditioned cucumber hypocotyl segments and enhance the activity of other H2O2 elicitors. Plant Physiol 117:1373–1380

Ferrándiz C, Gu Q, Martienssen R, Yanofsky MF (2000) Redundant regulation of meristem identity and plant architecture by FRUITFULL, APETALA1 and CAULIFLOWER. Development 127:725–734

Halgren TA (1996) Merck molecular force field. I. Basis, form, scope, parameterization, and performance of MMFF94. J Comp Chem 17:490–519

Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41:95–98

Hamilton JA (2004) Fatty acid interactions with proteins: what X-ray crystal and NMR solution structures tell us. Prog Lipid Res 43:177–199

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

Høgslund N, Radutoiu S, Krusell L, Voroshilova V, Hannah MA, Goffard N, Sanchez DH, Lippold F, Ott T, Sato S, Tabata S, Liboriussen P, Lohmann GV, Schauser L, Weiller GF, Udvardi MK, Stougaard J (2009) Dissection of symbiosis and organ development by integrated transcriptome analysis of lotus japonicus mutant and wild-type plants. PLoS ONE 4(8):e6556. doi:10.1371/journal.pone.0006556

Hoh F, Pons JL, Gautier MF, de Lamotte F, Dumas C (2005) Structure of a liganded type 2 non-specific lipid-transfer protein from wheat and the molecular basis of lipid binding. Acta Crystallogr D Biol Crystallogr 61:397–406

Horvath BM, Bachem CWB, Trindade LM, Oortwijn MEP, Visser RGF (2002) Expression analysis of a family of nsLTP genes tissue specifically expressed throughout the plant and during potato tuber life cycle. Plant Physiol 129:1494–1506

Jenks MA, Rashotte AM, Tuttle HA, Feldman KA (1996) Mutants in Arabidopsis thaliana altered in epicuticular wax and leaf morphology. Plant Physiol 110:377–385

Jones DT (1999) Protein secondary structure prediction based on position specific scoring matrices. J Mol Biol 292:195–202

Jorgensen WL, Chandrasekhar J, Madura JD, Impey RW, Klein ML (1983) Comparison of simple potential functions for simulating liquid water. J Chem Phys 79:926–935

José-Estanyol M, Gomis-Rüth FX, Puigdomènech P (2004) The eight-cysteine motif, a versatile structure in plant proteins. Plant Physiol Biochem 42:355–365

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 Environ 26:915–928

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

Kader JK (1997) Lipid transfer proteins: a puzzling family of plant proteins. Trends Plant Sci 2:66–70

Kandel S, Morant M, Benveniste I, Blee E, Werck-Reichhart D, Pinot F (2005) Cloning, functional expression, and characterization of CYP709C1, the first sub-terminal hydroxylase of long chain fatty acid in plants. Induction by chemicals and methyl jasmonate. J Biol Chem 280:35881–35889

Kielbowicz-Matuk A, Rey P, Rorat T (2008) Theorgan-dependent abundance of a Solanum lipid transfer proteinis up-regulated up on osmotic constraints and associated with cold acclimation ability. J Exp Bot 59:2191–2203

Kim TH, Park JH, Kim MC, Cho SH (2008) Cutin monomer induces expression of the rice OsLTP5 lipid transfer protein gene. J Plant Physiol 165:345–349

Kosma DK, Bourdenx B, Bernard A, Parsons EP, Lü S, Joubès J, Jenks MA (2009) The impact of water deficiency on leaf cuticle lipids of Arabidopsis. Plant Physiol 151:1918–1929

Kumar S, Tamura K, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics analysis and sequence alignment. Brief Bioinform 5:150–163

Laskoswki RA, MacArthur MW, Moss DS, Thorton JM (1993) PROCHECK: a program to check the stereo chemical quality of protein structures. J Appl Cryst 26:283–291

Lerche MH, Poulsen FH (1998) Solution structure of barley lipid transfer protein complexed with palmitate. Two different binding modes of palmitate in the homologous maize and barley nonspecific lipid transfer proteins. Protein Sci 7:2490–2498

Lerche MH, Kragelund BB, Bech LM, Poulsen FM (1997) Barley lipid-transfer protein complexed with palmitoyl CoA: the structure reveals a hydrophobic binding site that can expand to fit both large and small lipid-like ligands. Structure 5:291–306

Li H, Pinot F, Sauveplane V, Werck-Reichhart D, Diehl P, Schreiber L, Franke R, Zhang P, Chen L, Gao Y, Liang W, Zhang D (2010) Cytochrome P450 family member CYP704B2 catalyzes the {omega}-hydroxylation of fatty acids and is required for anther cutin biosynthesis and pollen exine formation in rice. Plant Cell 22:173–190

Liu PP, von Dahl CC, Park SW, Klessig DF (2011) Interconnection between methyl salicylate and lipid-based long-distance signaling during the development of systemic acquired resistance in Arabidopsis and tobacco. Plant Physiol 155:1762–1768

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using realtime quantitative PCR and the 2∆∆C (T) Method. Methods 25:402–408

Luo B, Xue XY, Hu WL, Wang LJ, Chen XY (2007) An ABC transporter gene of Arabidopsis thaliana, AtWBC11, is involved in cuticle development and prevention of organ fusion. Plant Cell Physiol 48:1790–1802

Luthy R, Bowie JU, Eisenberg D (1992) Assessment of protein models with three-dimensional profiles. Nature 356:83–85

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

Miyamoto K, Shimizu T, Mochizuki S, Nishizawa Y, Minami E, Nojiri H, Yamane H, Okada K (2012) Stress-induced expression of the transcription factor RERJ1 is tightly regulated in response to jasmonic acid accumulation in rice. Protoplasma. doi:10.1007/s00709-012-0400-z

Moons A, Keyser AD, Van Montagu M (1997) A group 3 LEA cDNA of rice, responsive to abscisic acid, but not to jasmonic acid, shows variety-specic differences in salt stress response. Gene 191:197–204

Munemasa S, Hossain MA, Nakamura Y, Mori IC, Murata Y (2011) The Arabidopsis calcium-dependent protein kinase, CPK6, functions as a positive regulator of methyl jasmonate signaling in guard cells. Plant Physiol 155:553–561

Nadakuduti SS, Pollard M, Kosma DK, Allen C Jr, Ohlrogge JB, Barry CS (2012) Pleiotropic phenotypes of the sticky peel mutant provide new insight into the role of CUTIN DEFICIENT2 in epidermal cell function in tomato. Plant Physiol 159:945–960

Nemethy G, Gibson KD, Palmer KA, Yoon CN, Paterlini G, Zagari A, Rumsey S, Scheraga HA (1992) Energy parameters in polypeptides.10. Improved geometrical parameters and non bonded interactions for use in the Ecepp/3 algorithm, with application to proline-containing peptides. J Phys Chem 96:6472–6484

Nieuwland J, Feron R, Huisman BAH, Fasolino A, Hilbers CW, Derksen J, Mariani C (2005) Lipid transfer proteins enhance cell wall extension in tobacco. Plant Cell 17:2009–2019

Ostergaard J, Vergnolle C, Schoentgen F, Kader JC (1993) Acyl-binding/lipid-transfer proteins from rape seedlings, a novel category of proteins interacting with lipids. Biochim Biophys Acta 13:109–117

Panikashvili D, Savaldi-Goldstein S, Mandel T, Yifhar T, Franke RB, Hofer R, Schreiber L, Chory J, Aharoni A (2007) The Arabidopsis DESPERADO/AtWBC11 transporter is required for cutin and wax secretion. Plant Physiol 145:1345–1360

Phillips JC, Braun R, Wang W, Gumbart J, Tajkhorshid E, Villa E, Chipot C, Skeel RD, Kalé L, Schulten K (2005) Scalable molecular dynamics with NAMD. J Comput Chem 26:1781–1802

Pighin JA, Zheng H, Balakshin LJ, Goodman IP, Western TL, Jetter R, Kunst L, Samuels AL (2004) Plant cuticular lipid export requires an ABC transporter. Science 306:702–704

Pii Y, Astegno A, Peroni E, Zaccardelli M, Pandolfini T, Crimi M (2009) The Medicago truncatula N5 gene encoding a root-specific lipid transfer protein is required for the symbiotic interaction with Sinorhizobium meliloti. Mol Plant Microbe Interact 22:1577–1587

Pinot F, Beneviste I, Salaün JP, Durst F (1998) Methyl jasmonate induces lauric acid omega-hydroxylase activity and accumulation of CYP94A1 transcripts but does not affect epoxide hydrolase activities in Vicia sativa seedlings. Plant Physiol 1181:1481–1486

Pons JL, de Lamotte F, Gautier MF, Delsuc MA (2003) Refined solution structure of a liganded type 2 wheat nonspecific lipid transfer protein. J Biol Chem 278:14249–14256

Pyee J, Kolattukudy PE (1995) The gene for the major cuticular wax-associated protein and three homologous genes from broccoli (Brassica oleracea) and their expression patterns. Plant J 7:49–59

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

Rizhsky L, Liang H, Shuman J, Shulaev V, Davletova S, Mittler R (2004) When defense pathways collide: the response of Arabidopsis to a combination of drought and heat stress. Plant Physiol 134:1683–1696

Sabala I, Elfstrand M, Farbos I, Clapham D, von Arnold S (2000) Tissue-specific expression of Pa18, a putative lipid transfer protein gene, during embryo development in Norway spruce (Piceaabies). Plant Mol Biol 42:461–478

Sali A, Blundell TL (1993) Comparative protein modelling by satisfaction of spatial restraints. J MolBiol 234:779–815

Sanchez DH, Lippold F, Redestig H, Hannah MA, Erban A, Krämer U, Kopka J, Udvardi MK (2008) Integrative functional genomics of salt acclimatization in the model legume Lotus japonicus. Plant J 53:973–987

Sato Sh, Nakamura Y, Kaneko T, Asamizu E, Kato T, Nakao M, Sasamoto S, Watanabe A, Ono A, Kawashima K, Fujishiro T, Katoh M, Kohara M, Kishida Y, Minami C, Nakayama S, Nakazaki N, Sawano Y, Hatano K, Miyakawa T, Komagata H, Miyauchi Y, Yamazaki H, Tanokura M (2008) Proteinase inhibitor from Ginkgo seeds is a member of the plant nonspecific lipid transfer protein gene family. Plant Physiol 146:1909–1919

Schlenkrich M, Brickman J, MacKerell AD, Karplus M (1996) Empirical potential energy function for phospholipids: criteria for parameter optimization and applications. In: Merz KM, Roux B (eds) Biological membranes: a molecular perspective from computation and experiment. Birkhauser, Boston, pp 31–81

Schnurr J, Shockey J, Browse J (2004) The acyl-CoA synthetase encoded by LACS2 is essential for normal cuticle development in Arabidopsis. Plant Cell 16:629–642

Schweizer P, Jeanguenat A, Whitacre D, Métraux J-P, Mösinger E (1996) Induction of resistance in barley against Erysiphegraminisfsphordei by free cutin monomers. Physiol Mol Plant Pathol 49:103–120

Shah J, Zeier J (2013) Long-distance communication and signal amplification in systemic acquired resistance. Front Plant Sci 4:30

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

Sippl MJ (1993) Recognition of errors in three-dimensional structures of proteins. Proteins 17:355–362

Suh MC, Samuels AL, Jetter R, Kunst L, Pollard M, Ohlrogge J, Beisson F (2005) Cuticular lipid composition, surface structure, and gene expression in Arabidopsis stem epidermis. Plant Physiol 139:1649–1665

Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

Tapia G, Morales-Quintana L, Inostroza L, Acuña H (2011) Molecular characterisation of Ltchi7, a gene encoding a Class III endochitinase induced by drought stress in Lotus spp. Plant Biol (Stuttg) 13:69–77

Tapia G, Acuña H, Inostroza L (2012) Plant drought tolerance: some genetics and agronomics relevant aspects for breeding in forage species. In: Neves DF, Sanz JD (eds) Droughts: new research. Nova Science Publishers, Hauppauge, pp 157–188

Thomson J, Higgins D, Gibson T (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4690

Vieira-Pires RS, Morais-Cabral JH (2010) 3(10) helices in channels and other membrane proteins. J Gen Physiol 136:585–592

Wallace AC, Laskowski RA, Thornton JM (1995) LIGPLOT: a program to generate schematic diagrams of protein–ligand interactions. Protein Eng 8:127–134

Yeats TH, Rose JKC (2008) The biochemistry and biology of extracellular plant lipid-transfer proteins (LTPs). Protein Sci 17:191–198

Yeats TH, Howe KJ, Matas AJ, Buda GJ, Thannhauser TW, Rose JK (2010) Mining the surface proteome of tomato (Solanum lycopersicum) fruit for proteins associated with cuticle biogenesis. J Exp Bot 61:3759–3771

Zhou J, Wang X, Jiao Y, Qin Y, Liu X, He K, Chen C, Ma L, Wang J, Xiong L, Zhang Q, Fan L, Deng XW (2007) Global genome expression analysis of rice in response to drought and high-salinity stresses in shoot, flag leaf, and panicle. Plant Mol Biol 63:591–608