Abstract
Very little is known about lipid transfer proteins from flax (Linum usitatissimum L.). In the present work, three genes encoding a lipid transfer protein (LTP) were isolated from flax, two of which encoded Type-1 and one Type-2 LTPs with molecular masses of about 9 and 7 kDa, respectively. The analysis of deduced amino acid sequence reveals that only Type 2 of the L. usitatissimum leaf specific LTP (LuLTP_Ls) had an N terminal signal peptide consisting of 23 amino acids. The phylogenetic analyses of LuLTP_Ls suggest their closest relatedness with respective proteins from Dimocarpus longan and Vitis vinifera. The gene expression analysis shows that LTP Type 1 genes, which include LuLTP_Ls1 and LuLTP_Ls3, were progressively expressed during leaf development, whereas LuLTP_Ls4 (Type 2) was expressed only at initial and terminal senescence stages of cotyledons. The results suggest that both types of LuLTP_Ls were differentially yet significantly expressed in cotyledons implicating their function in transport and scavenging lipidic skeletons for the benefit of other developing parts of the plant.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- EST:
-
expressed sequence tag
- GAL 1 :
-
galactose inducible 1
- His:
-
histidine
- LuLTP_Ls:
-
Linum usitatissimum lipid transfer protein
- ORF:
-
open reading frame
- RT-PCR:
-
reverse transacriptase - polymerase chain reaction
References
Altschul, S.F., Gish, W., Miller, W., Myers, E.W., Lipman, D.J.: Basic local alignment search tool. — J. mol. Biol. 215: 403–410, 1990.
Browse, J., Warwick, N., Somerville, C.R., Slack, C.R.: Fluxes through the prokaryotic and eukaryotic pathways of lipid synthesis in the ‘16:3’ plant Arabidopsis thaliana. — Biochem. J. 235: 25–31, 1986.
Buhot, N., Douliez, J.P., Jacquemard, A., Marion, D., Tran, V., Maume, B.F., Milat, M.L., Ponchet, M., Mikes, V., Kader, J.C., Blein, J.P.: A lipid transfer protein binds to a receptor involved in the control of plant defence responses. — FEBS Lett. 509: 27–30, 2001.
Cameron, K.D., Teece, M.A., Smart, L.B.: 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, 2006.
Cammue, B. P. A., Thevissen, K., Hendriks, M., Eggermont, K., Goderis, I. J., Proost, P., Broekaert, W. F.: A potent antimicrobial protein from onion seeds showing sequence homology to plant lipid transfer proteins. — Plant. Physiol. 109: 445–455, 1995.
Charvolin, D., Douliez, J., Marion, D., Cohen-Addad, C., Pebay-Peyroula, E: The crystal structure of a wheat nonspecific lipid transfer protein (ns-LTP1) complexed with two molecules of phospholipid at 2.1 Å resolution. - Eur. J. Biochem. 264: 562–568, 1999.
Douliez, J.P., Michon, T., Elmorjani, K., Marion, D.: 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, 2000.
Edqvist, J., Farbos, I.: Characterization of germination-specific lipid transfer proteins from Euphorbia lagascae. — Planta 215: 41–50, 2002.
Eklund, D.M., Edqvis, J.: Localization of nonspecific lipid transfer proteins correlate with programmed cell death responses during endosperm degradation in Euphorbia lagascae seedlings. — J. Plant. Physiol. 132: 1249–1259, 2003.
Fleming, A.J., Mandel, T., Hofman, S., Sterk, P., De Vries, S.C., Kuhlemeier, C.: Expression pattern of a tobacco lipid transfer protein gene within the shoot apex. — Plant. J. 2: 855–862, 1992.
Han, G.W., Lee, J.Y., Song, H.K., Chang, C., Min, K., Moon, J., Shin, D.H., Kopka, M.L., Sawaya, M.R., Yuan, H.S.: Structure 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, 2001.
Hortensteiner, S., Feller, U.: Nitrogen metabolism and remobilization during senescence. — J. exp. Bot. 53: 927–937, 2002.
Kader, J.C.: Lipid-transfer protein in plants. — Annu. Rev. Plant Physiol. Plant. mol. Biol. 47: 627–654, 1996.
Kim, T.H., Kim, M.C., Park, J.H., Han, S.S., Kim, B.R., Moon, B.Y., Cho, S.H.: Differential expression of rice lipid transfer protein gene (LTP) classes in response to abscisic acid, salt, salicylic acid, and the fungal pathogen Magnaporthe grisea. — J. Plant. Biol. 49: 371–375, 2006.
Laemmli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. — Nature 227: 680–685, 1970.
Lascombe, M.B., Bakan, B., Buhot, N., Marion, D., Blein, J.P., Larue, V., Lamb, C., Prange, T.: The structure of “defective in induced resistance” protein of Arabidopsis thaliana, DIR1, reveals a new type of lipid transfer protein. — Protein. Sci. 17: 1522–1530, 2008.
Lee, J.Y., Min, K., Cha, H., Shin, D.H., Hwang, K.Y., Suh, S.W.: Rice nonspecific lipid transfer protein: the 1.6 Å crystal structure in the unliganded state reveals a small hydrophobic cavity. — J. mol. Biol. 276: 437–448, 1998.
Liu, Y.J., Samuel, D., Lin, C.H., Liu, P.C.: Purification and characterization of a novel 7-kDa non-specific lipid transfer protein-2 from rice (Oryza sativa). — Biochem. biophys. Res. Commun. 294: 535–540, 2002.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin phenol reagent. — J. biol. Chem. 193: 265–275, 1951.
Molina, A., Garcia-Olmedo, F.: Developmental and pathogeninduced expression of three barley genes encoding lipid transfer proteins. — Plant. J. 4: 983–991, 1993.
Molina, A., Segura, A., Garcia-Olmedo, F.: Lipid transfer proteins (nsLTPs) from barley and maize leaves are potent inhibitors of bacterial and fungal plant pathogens. — FEBS. Lett. 316: 119–22, 1993.
Nooden, L.D., Guiamet, J.J.: Regulation of assimilation and senescence by the fruit in monocarpic plants. — Physiol. Plant. 77: 267–274, 1989.
Pasquato, N., Berni, R., Folli, C., Folloni, S., Cianci, M., Pantano, S., Zanotti, G.: Crystal structure of peach Pru p 3, the prototypic member of the family of plant non-specifc lipid transfer protein pan-allergens. — J. mol. Biol. 356: 684–694, 2006.
Petersen, T.N., Brunak, S., Von Heijne, G., Nielsen, H.: Signal P 4.0: discriminating signal peptides from transmembrane regions. — Natur. Methods 8: 785–786, 2011.
Quirino, B.F., Noh, Y.S., Himelblau, E., Amasino, R.M.: Molecular aspects of leaf senescence. — Trends Plant. Sci. 5: 278–282, 2000.
Salcedo, G., Sanchez-Monge, R., Barber, D., Diaz-Perales, A.: Plant nonspecific lipid transfer proteins: an interface between plant defence and human allergy. — Biochim. biophys. Acta 1771: 781–791, 2007.
Schmit, M., Simpson, D., Gietl, C.: Programmed cell death in castor bean endosperm is associated with the accumulation and release of a cysteine endopeptidase from ricinosomes. — Proc. nat. Acad. Sci. USA 96: 14159–14164, 1999.
Segura, A., Moreno, M., Garcia-Olmedo, F.: Purification and antipathogenic activity of lipid transfer proteins (LTPs) from the leaves of Arabidopsis and spinach. — FEBS Lett. 332: 243–246, 1993.
Shahri, W., Tahir, I.: Physiological and biochemical changes associated with flower development and senescence in Consolida ajacis Nieuwl cv.Violet blue. — Front. Agr. China 5: 201–208, 2011.
Tamura, K., Dudley, J., Nei, M., Kumar, S.: MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. — Mol. Biol. Evol. 24: 1596–1599, 2007.
Terras, F.R.G., Goderis, I.J., Van Leuven, F., Van der Leyden, J., Camimue, B.P.A., Broekaert, W.F.: In vitro antifungal activity of a radish (Raphanus sativus L.) seed protein homologous to nonspecific lipid transfer proteins. — Plant. Physiol. 100: 1055–1058, 1992.
Wang, S.Y., Wu, J.H., Ng, T.B., Ye, X.Y., Rao, P.F.: A non-specific lipid transfer protein with antifungal and antibacterial activities from the mung bean. — Peptides 25: 1235–1242, 2004.
Zaman, U., Abbasi, A.: Isolation, purification and characterization of a nonspecific lipid transfer protein from Cuminum cyminum. — Phytochemistry 70: 979–987, 2009.
Zottich, U., Da, Cunha. M., Carvalho, A.O., Dias, G.B., Silva, N.C., Santos, I.S., do Nacimento, V.V., Miguel E.C., Machado, O.L., Gomes, V.M.: Purification, biochemical characterization and antifungal activity of a new lipid transfer protein (LTP) from Coffea canephora seeds with α-amylase inhibitor properties. — Biochim. biophys. Acta 1810: 375–383, 2011.
Author information
Authors and Affiliations
Corresponding author
Additional information
Acknowledgments: Financial assistance to the Indian side by the Department of Biotechnology, Govt. of India and Genome Canada and Genome Prairie funding through the TUFGEN project to Raju Datla is acknowledged.
Rights and permissions
About this article
Cite this article
Mhaske, V.A., Datla, R., Qiu, S. et al. Isolation and characterization of genes encoding lipid transfer proteins in Linum usitatissimum . Biol Plant 60, 285–291 (2016). https://doi.org/10.1007/s10535-016-0592-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10535-016-0592-8