Molecular characterization and subcellular localization of salt-inducible lipid transfer proteins in rice (2024)

Biologia plantarum 61:501-510, 2017|DOI:10.1007/s10535-016-0671-x

Molecular characterization and subcellular localization of salt-inducible lipid transfer proteins in rice

K.-C. Lin¹, T.-M. Wu^1,2, N. N. P. Chandrika¹, S.-J. Chou³, C.-Y. Hong^1,*

¹Department of Agricultural Chemistry, National Taiwan University, Taipei, Taiwan

²Department of Aquaculture, National Pingtung University of Science and Technology, Pingtung, Taiwan

³Institute of Plant and Microbial Biology, Academia Sinica, Taipei, Taiwan

Rice (Oryza sativa L.) is a salt-sensitive species. Salt stress can cause injury to the plant cellular membrane. Plant lipid transfer proteins (LTPs) are abundant lipid binding proteins that are important in membrane vesicle biogenesis and trafficking, however, the biological importance of LTPs on salt-stress response in rice remains unclear. Therefore, salt-responsive rice LTPs were identified and characterized in this study. Microarray analysis showed seven genes positively regulated by salinity, including five Ltp genes (LtpII.3, LtpII.5, LtpII.6, LtpV.1, and LtpV.2) and two Ltp-like (LtpL; LtpL1, and LtpL2) genes. Amino acid alignment revealed that all these Ltp and LtpL genes contained the N-terminal signal peptide. Apart from LtpL1, all salt-inducible Ltp genes had the conserved eight cysteine residue motifs backbone. Verification of gene expression to different stimuli in rice seedlings revealed that salt-regulated Ltp genes differentially responded to drought, cold, H₂O₂, abscisic acid (ABA) and CaCl₂. Furthermore, the expression of Ltp and LtpL genes was tissue-specifically regulated by ABA-dependent and independent pathway. In silico analysis of a 1.5-kb 5'-upstream region of these genes showed regulatory cis-elements associated with ABA, calcium, and cold/drought responses. Three LtpII subfamily genes, including LtpII.3, LtpII.5, and LtpII.6, were strictly expressed in flowers and seeds, and LtpIII.1 mRNA strongly accumulated in stem tissue. Subcellular localization analysis of LTP-DsRed fusion proteins revealed that the five LTPs and two LTPLs localized at the endoplasmic reticulum. The results provide new clues to further understanding the biological functions of Ltp genes.

Keywords: abscisic acid; calcium; cold; drought; gene expression; Oryza sativa; subcellular localization; salinity

Subjects: lipid transfer protein; salinity; abscsic acid; calcium; cold stress; drought; gene expression; subcellular localization; rice

Received: February 5, 2016; Revised: June 17, 2016; Accepted: July 12, 2016; Published: September 1, 2017 Show citation

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1. Ahmad, P., Hakeem, K.R., Kumar, A., Ashraf, M., Akram, N.A.: Salt-induced changes in photosynthetic activity and oxidative defense system of three cultivars of mustard (Brassica juncea L.). - Afr. J. Biotechnol. 11: 2694-2703, 2012. Go to original source...
2. Beisson, F., Koo, A.J., Ruuska, S., Schwender, J., Pollard, M., Thelen, J.J., Paddock, T., Salas, J.J., Savage, L., Milcamps, A., Mhaske, V.B., Cho, Y., Ohlrogge, J.B.: Arabidopsis genes involved in acyl lipid metabolism. A 2003 census of the candidates, a study of the distribution of expressed sequence tags in organs, and a web-based database. - Plant Physiol. 132: 681-697, 2003. Go to original source...
3. Boutrot, F., Chantret, N., Gautier, M.F.: 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: 86-104, 2008. Go to original source...
4. Charvolin, D., Douliez, J.P., 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. Go to original source...
5. Christensen, A.H., Quail, P.H.: Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. - Transgen. Res. 5: 213-218, 1996. Go to original source...
6. Crooks, G.E., Hon, G., Chandonia, J.M., Brenner, S.E.: WebLogo: a sequence logo generator. - Genome Res. 14:1188-1190, 2004. Go to original source...
7. Dionisio-Sese, M.L., Tobita, S.: Antioxidant responses of rice seedlings to salinity stress. - Plant Sci. 135:1-9, 1998. Go to original source...
8. Diz, M.S., Carvalho, A.O., Ribeiro, S.F., Da Cunha, M., Beltramini, L., Rodrigues, R., Nascimento, V.V., Machado, O.L., Gomes, V.M.: Characterisation, immunolocalisation and antifungal activity of a lipid transfer protein from chili pepper (Capsicum annuum) seeds with novel α-amylase inhibitory properties. - Physiol. Plant. 142: 233-246, 2011. Go to original source...
9. Edstam, M.M., Laurila, M., Höglund, A., Raman, A., Dahlström, K.M., Salminen, T.A., Edqvist, J., Blomqvist, K: Characterization of the GPI-anchored lipid transfer proteins in the moss Physcomitrella patens. - Plant Physiol. Biochem. 75: 55-69, 2014. Go to original source...
10. Edstam, M.M., Viitanen, L., Salminen, T.A., Edqvist, J.: Evolutionary history of the non-specific lipid transfer proteins. - Mol. Plant 4: 947-964, 2011. Go to original source...
11. Emanuelsson, O., Nielsen, H., Von Heijne, G.: ChloroP, a neural network-based method for predicting chloroplast transit peptides and their cleavage sites. - Protein Sci. 8: 978-984, 1999. Go to original source...
12. Gao, G., Jin, L.P., Xie, K.Y., Qu, D.Y.: The potato StLTPa7 gene displays a complex Ca-associated pattern of expression during the early stage of potato-Ralstonia solanacearum interaction. - Mol. Plant Pathol. 10: 15-27, 2009. Go to original source...
13. García-Garrido, J.M., Menossi, M., Puigdoménech, P., Martínez-Izquierdo, J.A., Delseny, M. Characterization of a gene encoding an abscisic acid-inducible type-2 lipid transfer protein from rice. - FEBS Lett. 428: 193-199, 1998. Go to original source...
14. Gill, S.S., Tuteja, N.: Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. - Plant Physiol. Biochem. 48: 909-930, 2010. Go to original source...
15. Grattan, S.R., Grieve, C.M.: Mineral nutrient acquisition and response by plants grown in saline environments. - In: Pessarakli, M. (ed.): Handbook of Plant and Crop Stress. Pp. 203-229. Marcel Dekker, New York 1999. Go to original source...
16. Gueta-Dahan, Y., Yaniv, Z., Zilinskas, B.A., Ben-Hayyim, G.: Salt and oxidative stress: similar and specific responses and their relation to salt tolerance in citrus. - Planta 203: 460-469, 1997. Go to original source...
17. Guiderdoni, E., Cordero, M.J., Vignols, F., García-Garrido, J.M., Lescot, M., Tharreau, D., Meynard, D., Ferrière, N., Notteghem, J.L., Delseny, M.: Inducibility by pathogen attack and developmental regulation of the rice Ltp1 gene. - Plant mol. Biol. 49: 683-699, 2002. Go to original source...
18. Guo, L., Yang, H., Zhang, X., Yang S.: Lipid transfer protein 3 as a target of MYB96 mediates freezing and drought stress in Arabidopsis. - J. exp. Bot. 64: 1755-1767, 2013. Go to original source...
19. Higo, K., Ugawa, Y., Iwamoto, M., Korenaga, T.: Plant cis-acting regulatory DNA elements (PLACE) database. - Nucl. Acids Res. 27: 297-300, 1999. Go to original source...
20. Heinemann, B., Andersen, K.V., Nielsen, P.R., Bech, L.M., Poulsen, F.M.: Structure in solution of a four-helix lipid binding protein. - Protein Sci. 5: 13-23, 1996. Go to original source...
21. Holmberg, N., Bülow, L.: Improving stress tolerance in plants by gene transfer. - Trends Plant Sci. 3: 61-66, 1998. Go to original source...
22. Jang, C.S., Lee, H.J., Chang, S.J., Seo, Y.W.: Expression and promoter analysis of the TaLTP1 gene induced by drought and salt stress in wheat (Triticum aestivum L.). - Plant Sci. 167: 995-1001, 2004. Go to original source...
23. Jose-Estanyol, M., Gomis-Ruth, F.X., Puigdomenech, P.: The eight-cysteine motif, a versatile structure in plant proteins. - Plant Physiol. Biochem. 42: 355-365, 2004. Go to original source...
24. Kader, J.C.: Lipid-transfer proteins in plants. - Annu. Rev. Plant Physiol. Plant mol. Biol. 47: 627-654, 1996. Go to original source...
25. Kader, J.C.: Lipid transfer proteins: a puzzling family of plant proteins. - Trends Plant Sci. 2: 66-70, 1997. Go to original source...
26. Kaplan, B., Davydov, O., Knight, H., Galon, Y., Knight, M.R., Fluhr, R., Fromm, H.: Rapid transcriptome changes induced by cytosolic Ca2+ transients reveal ABRE-related sequences as Ca2+-responsive cis elements in Arabidopsis. - Plant Cell 18: 2733-2748, 2006. Go to original source...
27. Kawahara, Y., De la Bastide, M., Hamilton, J.P., Kanamori, H., McCombie, W.R., Ouyang, S., Schwartz, D.C., Tanaka, T., Wu, J., Zhou, S., Childs, K.L., Davidson, R.M., Lin, H., Quesada-Ocampo, L., Vaillancourt, B., Sakai, H., Lee, S.S., Kim, J., Numa, H., Itoh, T., Buell, C.R., Matsumoto, T.: Improvement of the Oryza sativa Nipponbare reference genome using next generation sequence and optical map data. - Rice 6: 4, 2013. Go to original source...
28. Kim, T.H., Park, J.H., Kim, M.C., Cho, SH: cutin monomer induces expression of the rice OsLTP5 lipid transfer protein gene. - J. Plant Physiol. 165: 345-349, 2008. Go to original source...
29. Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., Lopez, R., Thompson, J.D., Gibson, T.J., Higgins, D.G.: Clustal W and Clustal X version 2.0. - Bioinformatics 23: 2947-2948, 2007. Go to original source...
30. Liu, F., Zhang, X., Lu, C., Zeng, X., Li, Y., Fu, D., Wu, G.: Non-specific lipid transfer proteins in plants: presenting new advances and an integrated functional analysis. - J. exp. Bot. 66: 5663-5681, 2015. Go to original source...
31. Liu, X., Shangguan, Y., Zhu, J., Lu, Y., Han, B.: The rice OsLTP6 gene promoter directs anther-specific expression by a combination of positive and negative regulatory elements. - Planta 238: 845-857, 2013. Go to original source...
32. Madrid, S.M.: The barley lipid transfer protein is targeted into the lumen of the endoplasmic reticulum. - Plant Physiol. Biochem. 29: 659-703, 1991.
33. Matz, M.V., Fradkov, A.F., Labas, Y.A., Savitsky, A.P., Zaraisky, A.G., Markelov, M.L., Lukyanov S, A.: Fluorescent proteins from nonbioluminescent Anthozoa species. - Nat. Biotechnol. 17: 969-973, 1999. Go to original source...
34. Mittler, R.: Oxidative stress, antioxidants and stress tolerance. - Trends Plant Sci. 7: 405-410, 2002. Go to original source...
35. Mittova, V., Guy, M., Tal, M., Volokita, M.: Salinity upregulates the antioxidative system in root mitochondria and peroxisomes of the wild salt-tolerant tomato species Lycopersicon pennellii. - J. exp. Bot. 55: 1105-1113, 2004. Go to original source...
36. Munns, R., Tester, M.: Mechanisms of salinity tolerance. - Annu. Rev. Plant Biol. 59: 651-681, 2008. Go to original source...
37. Pagnussat, L., Burbach, C., Baluska, F., De la Canal, L.: An extracellular lipid transfer protein is relocalized intracellularly during seed germination. - J. exp. Bot. 63: 6555-6563, 2012. Go to original source...
38. Potocka, I., Baldwin, T.C., Kurczynska, E.U.: Distribution of lipid transfer protein 1 (LTP1) epitopes associated with morphogenic events during somatic embryogenesis of Arabidopsis thaliana. - Plant Cell Rep. 31: 2031-2045, 2012. Go to original source...
39. Safi, H., Saibi, W., Alaoui, M.M., Hmyene, A., Masmoudi, K., Hanin, M., Brini, F.: A wheat lipid transfer protein (TdLTP4) promotes tolerance to abiotic and biotic stress in Arabidopsis thaliana. - Plant Physiol. Biochem. 89: 64-75, 2015. Go to original source...
40. Salama, K.H.A., Mansour, M.M.F., Ali, F.Z.M., Abou-Hadid, A.F.: NaCl-induced changes in plasma membrane lipids and proteins of Zea mays L. cultivars in their response to salinity. - Acta. Physiol. Plant. 29: 351-359, 2007. Go to original source...
41. Seo, P.J., Lee, S.B., Suh, M.C., Park, M.J., Go, Y.S., Park, C.M.: The MYB96 transcription factor regulates cuticular wax biosynthesis under drought conditions in Arabidopsis. - Plant Cell 23:1138-1152, 2011. Go to original source...
42. Shin, D.H., Lee, J.Y., Hwang, K.Y., Kim, K.K., Suh, S.W.: High resolution crystal structure of the nonspecific lipidtransfer protein from maize seedlings. - Structure 3: 189-199, 1995. Go to original source...
43. Silverstein, K.A., Moskal, W.A.J., Wu, H.A., Underwood, B.A., Graham, M.A., Town, C.D., Van den Bosch, K.A.: Small cysteine-rich peptides resembling antimicrobial peptides have been under-predicted in plants. - Plant J. 51: 262-280, 2007. Go to original source...
44. Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., Kumar, S.: MEGA 5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. - Mol. Biol. Evol. 28: 2731-2739, 2011. Go to original source...
45. Tapia, G., Morales-Quintana, L., Parra, C., Berbel, A., Alcorta, M.: 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 mol. Biol. 82: 485-501, 2013. Go to original source...
46. Torres-Schumann, S., Godoy, J.A., Pintor-Toro, J.A.: A probable lipid transfer protein gene is induced by NaCl in stems of tomato plants. - Plant mol. Biol. 18: 749-757, 1992. Go to original source...
47. Tsuboi, S., Osafune, T., Tsugeki, R., Nishimura, M., Yamada, M.: Nonspecific lipid transfer protein in castor bean cotyledon cells: subcellular localization and a possible role in lipid metabolism. - J. Biochem. 111: 500-508, 1992. Go to original source...
48. Vignols, F., Lund, G., Pammi, S., Tremousaygue, D., Grellet, F., Kader, J.C., Puigdomenech, P., Delseny, M.: Characterization of a rice gene coding for a lipid transfer protein. - Gene 142: 265-270, 1994. Go to original source...
49. Vignols, F., Wigger, M., García Garrido, J.M., Grellet, F., Kader, J.C., Delseny, M.: Rice lipid transfer protein (LTP) genes belong to a complex multigene family and are differentially regulated. - Gene 195: 177-186, 1997. Go to original source...
50. Wahid, A., Gelani, S., Ashraf, M., Foolad, M.: Heat tolerance in plants: an overview. - Environ. exp. Bot. 61: 199-223, 2007. Go to original source...
51. Wang, C., Xie, C., Chi, F., Hu, W., Mao, G., Sun, D., Li, C., Sun, Y.: BcLTP, a novel lipid transfer protein in Brassica chinensis, may secrete and combine extracellular CaM. - Plant Cell Rep. 27: 159-169, 2008. Go to original source...
52. Wang, F., Zang, X., Kabir, M.R., Liu, K., Liu, Z., Ni, Z., Yao, Y., Hu, Z., Sun, Q., Peng, H.A.: Wheat lipid transfer protein 3 could enhance the basal thermotolerance and oxidative stress resistance of Arabidopsis. - Gene 550: 18-26, 2014. Go to original source...
53. Wang, H.W., Hwang, S.G., Karuppanapandian, T., Liu, A., Kim, W., Jang, C.S.: Insight into the molecular evolution of nonspecific lipid transfer proteins via comparative analysis between rice and sorghum. - DNA Res. 19: 179-194, 2012. Go to original source...
54. Wu, T.M., Lin, K.C., Liau, W.S., Chao, Y.Y., Yang, L.H., Chen, S.Y., Lu, C.A., Hong, C.Y.: A set of GFP-based organelle marker lines combined with DsRed-based gateway vectors for subcellular localization study in rice (Oryza sativa, L.). - Plant mol. Biol. 90: 107-115, 2015. Go to original source...
55. Yeats, T.H., Rose, J.K.C.: The biochemistry and biology of extracellular plant lipid-transfer proteins (LTPs). - Protein Sci. 17: 191-198, 2008. Go to original source...
56. Yoo, S.D., Cho, Y.H., Sheen, J.: Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. - Nat. Protocols 2: 1565-1572, 2007. Go to original source...
57. Yoshida, S., Forno, D.A., co*ck, J.H., Gomez, K.A.: Laboratory Manual for Physiological Studies of Rice. 2nd Ed. - The International Rice Research Institute, Los Baños 1972.
58. Yu, G., Hou, W., Du, X., Wang, L., Wu, H., Zhao, L., Kong, L., Wang, H.: Identification of wheat non-specific lipid transfer proteins involved in chilling tolerance. - Plant Cell Rep. 33: 1757-1766, 2014. Go to original source...
59. Yu, K., Soares, J.M., Mandal, M.K., Wang, C., Chanda, B., Gifford, A.N., Fowler, J.S., Navarre, D., Kachroo, A., Kachroo, P.: A feedback regulatory loop between G3P and lipid transfer proteins DIR1 and AZI1 mediates azelaicacid-induced systemic immunity. - Cell Rep. 3: 1266-1278, 2013. Go to original source...
60. Yubero-Serrano, E.M., Moyano, E., Medina-Escobar, N., Muñoz-Blanco, J., Caballero, J.L.: Identification of a strawberry gene encoding a non-specific lipid transfer protein that responds to ABA, wounding and cold stress. - J. exp. Bot. 54: 1865-1877, 2003. Go to original source...
61. Zamani, S., Bybordi, A., Khorshidi, A., Nezami, T.: Effects of NaCl salinity levels on lipids and proteins of canola (Brassica napus L.) cultivars. - Adv. environ. Biol. 4: 397-403, 2010.
62. Zhang, D., Liang, W., Yin, C., Zong, J., Gu, F.: OsC6, encoding a lipid transfer protein, is required for postmeiotic anther development in rice. - Plant Physiol. 154: 49-162, 2010. Go to original source...
63. Zhang, Y., Su, J., Duan, S., Ao, Y., Dai, J., Liu, J., Wang, P., Li, Y., Liu, B., Feng, D., Wang, J., Wang, H.: A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. - Plant Methods 7: 30-43, 2011. Go to original source...