Abstract
This study was to investigate the responses of phospholipase D (PLD) and lipoxygenase (LOX) to mechanical wounding in postharvest cucumber (Cucumis sativus L. cv. Biyu-2) fruits. Membrane-associated Ca2+ content, activities and gene expression of PLD and LOX, and contents of phosphatidylcholine (PC), phosphatidylinositol (PI), and phosphatidic acid (PA) were determined in cucumber fruits following mechanical wounding. Results show that PLD and LOX activities increased with the PLD and LOX mRNAs which are upregulated upon wounding, while membrane-associated Ca2+ content decreased. Accompanying with the increase of PLD and LOX activities, accumulation of PA and losses of PC and PI were observed in all fruits, but there were differences of degrees between wounded and control fruits. Results suggest that PLD and LOX might be the main hydrolytic enzymes of phospholipids in postharvest cucumber fruits participating in the mechanical wounding injury. The activation of PLD and LOX might be the result of gene expression, which could be stimulated by the Ca2+ flowing from the membrane to the cytoplasm upon receiving the wounding signals.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bargmann, B.O.R., Munnik, T., 2006. The role of phospholipase D in plant stress responses. Current Opinion in Plant Biology, 9(5):515–522. [doi:10.1016/j.pbi.2006.07.011]
Bargmann, B.O.R., Laxalt, A.M., Ter Riet, B., Testerink, C., Merquiol, E., Mosblech, A., Leon-Reyes, A., Pieterse, C.M.J., Haring, M.A., Heilmann, I., et al., 2009. Reassessing the role of phospholipase D in the Arabidopsis wounding response. Plant Cell & Environment, 32(7): 837–850. [doi:10.1111/j.1365-3040.2009.01962.x]
Bush, D.S., 1995. Calcium regulation in plant-cells and its role in signaling. Annual Review of Plant Physiology and Plant Molecular Biology, 46(1):95–122. [doi:10.1146/annurev.pp.46.060195.000523]
Conconi, A., Miguel, M., Browse, J.A., Ryan, C.A., 1996. Intracellular levels of free linolenic and linoleic acids increase in tomato leaves in response to wounding. Plant Physiology, 111(3):797–803.
Creelman, R.A., Tierney, M.L., Mullet, J.E., 1992. Jasmonic acid/methyl jasmonate accumulate in wounded soybean hypocotyls and modulate wound gene expression. Proceedings of the National Academy of Sciences, 89(11): 4938–4941. [doi:10.1073/pnas.89.11.4938]
de Bruxelles, G.L., Roberts, M.R., 2001. Signals regulating multiple responses to wounding and herbivores. Critical Reviews in Plant Sciences, 20(5):487–521. [doi:10.1016/S0010-8545(02)00029-2]
Deschene, A., Paliyath, G., Lougheed, E.C., Dumbroff, E.B., Thompson, J.E., 1991. Membrane deterioration during postharvest storage of broccoli florets: modulation by temperature and controlled atmosphere storage. Postharvest Biology and Technology, 1(1):19–31. [doi:10.1016/0925-5214(91)90016-5]
de Vrije, T., Munnik, T., 1997. Activation of phospholipase D by calmodulin antagonists and mastoparan in carnation petals. Journal of Experimental Botany, 48(9):1631–1637. [doi:10.1093/jxb/48.9.1631]
Durigan, M.F.B., Mattiuz, B.H., 2007. Effect of mechanical injuries on the quality of watermelon stored at room temperature. Horticultura Brasileira, 25(2):296–300. [doi: 10.1590/S0102-05362007000200033]
Feng, J.L., Wang, K., Liu, X., Chen, S.N., Chen, J.S., 2009. The quantification of tomato microRNAs response to viral infection by stem-loop real-time RT-PCR. Gene, 437(1–2):14–21. [doi:10.1016/j.gene.2009.01.017]
Gil-Salas, F.M., Colyer, A., Boonham, N., Cuadrado, I.M., Janssen, D., 2009. Resistance screening against cucumber vein yellowing virus using a real-time (TaqMan) RT-PCR assay in cucumber (Cucumis sativus). Crop Protection, 28(1):109–112. [doi:10.1016/j.cropro.2008.09.005]
Khatoon, H., Talat, S., Younus, H., 2008. Phospholipase D from Allium sativum bulbs: a highly active and thermal stable enzyme. International Journal of Biological Macromolecules, 42(4):380–385. [doi:10.1016/j.ijbiomac.2008.01.007]
Lee, S., Suh, S., Kim, S., Crain, R.C., Kwak, J.M., Nam, H.G., Lee, Y., 1997. Systemic elevation of phosphatidic acid and lysophospholipid levels in wounded plants. The Plant Journal, 12(3):547–556. [doi:10.1046/j.1365-313X.1997.00547.x]
Lee, S.H., Ahn, S.J., Im, Y.J., Cho, K., Chung, G.C., Cho, B.H., Han, O., 2005. Differential impact of low temperature on fatty acid unsaturation and lipoxygenase activity in fig leaf gourd and cucumber roots. Biochemical and Biophysical Research Communications, 330(4):1194–1198. [doi:10.1016/j.bbrc.2005.03.098]
Lein, W., Saalbach, G., 2001. Cloning and direct G-protein regulation of phospholipase D from tobacco. Biochimica et Biophysica Acta (BBA)-Molecular and Cell Biology of Lipids, 1530(2–3):172–183. [doi:10.1016/S1388-1981(00)00182-7]
Li, L., Jiao, X.Z., 1980. Measurement of protein content by the agency of protein stain Coomassie Brilliant Blue G250. Plant Physiology Communications, 6:52–55.
Livak, K.J., Thomas, D., Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 −ΔΔCr method. Methods, 25(4):402–408. [doi:10.1006/meth.2001.1262]
Mao, L.C., Wang, G.Z., Zhu, C.G., Pang, H.Q., 2007a. Involvement of phospholipase D and lipoxygenase in response to chilling stress in postharvest cucumber fruits. Plant Science, 172(2):400–405. [doi:10.1016/j.plantsci.2006.10.002]
Mao, L.C., Pang, H.Q., Wang, G.Z., Zhu, C.G., 2007b. Phospholipase D and lipoxygenase activity in cucumber fruit in response to chilling stress. Postharvest Biology and Technology, 44(1):42–47. [doi:10.1016/j.postharvbio.2006.11.009]
Martínez-Romero, D., Serrano, M., Riquelme, F., 1999. Effects of postharvest putrescine and calcium treatments on reducing mechanical damage and polyamines and ABA levels during lemon storage. Journal of the Science of Food and Agriculture, 79(12):1589–1595. [doi:10.1002/(SICI)1097-0010(199909)79:12<1589::AID-JSFA403>3.0.CO;2-J]
Narvaez-Vasquez, J., Florin-Christensen, J., Ryan, C.A., 1999. Positional specificity of a phospholipase A activity induced by wounding, systemin and oligosaccharide elicitors in tomato leaves. The Plant Cell Online, 11(11): 2249–2260. [doi:10.1105/tpc.11.11.2249]
Paliyath, G., Thompson, J.E., 1987. Calcium and calmodulin regulated breakdown of phospholipid by microsomal membranes from bean cotyledons. Plant Physiology, 83(1):63–68. [doi:10.1104/pp.83.1.63]
Paliyath, G., Droillard, M.J., 1992. The mechanisms of membrane deterioration and disassembly during senescence. Plant Physiology and Biochemistry, 30:789–812.
Pinhero, R.G., Paliyath, G., Yada, R.Y., Murr, D.P., 1998. Modulation of phospholipase D and lipoxygenase activities during chilling: relation to chilling tolerance of maize seedlings. Plant Physiology and Biochemistry, 36(3): 213–224. [doi:10.1016/S0981-9428(97)86878-7]
Ryu, S.B., Wang, X., 1996. Activation of phospholipase D and the possible mechanism of activation in wound-induced lipid hydrolysis in castor bean leaves. Biochimica et Biophysica Acta (BBA)-Lipids and Lipid Metabolism, 1303(3):243–250. [doi:10.1016/0005-2760(96)00096-3]
Saltveit, M.E., 2000. Wound-induced changes in phenolic metabolism and tissue browning are altered by heatshock. Postharvest Biology and Technology, 21(1):61–69. [doi: 10.1016/S0925-5214(00)00165-4]
Saravitz, D.M., Siedow, J.N., 1996. The differential expression of wound-inducible lipoxygenase gene in soybean leaves. Plant Physiology, 110(1):287–299. [doi:10.1104/pp.110.1.287]
Todd, J.F., Paliyath, G., Thompson, J.E., 1990. Characteristics of a membrane associated lipoxygenase in tomato fruit. Plant Physiology, 94(3):1225–1232. [doi:10.1104/pp.94.3.1225]
van Linden, V., Sila, D.N., Duvetter, T., Baerdemaeker, J.D., Hendrickx, M., 2008. Effect of mechanical impact-bruising on polygalacturonase and pectinmethylesterase activity and pectic cell wall components in tomato fruit. Postharvest Biology and Technology, 47(1):98–106. [doi: 10.1016/j.postharvbio.2007.06.006]
Wang, X.M., 2000. Multiple forms of PLD in plant: the gene family, catalytic and regulatory properties, and cellular functions. Progress in Lipid Research, 39(2):109–149. [doi:10.1016/S0163-7827(00)00002-3]
Wang, X.M., 2005. Regulatory functions of phospholipase D and phosphatidic acid in plant growth, development and stress responses. Plant Physiology, 139(2):566–573. [doi: 10.1104/pp.105.068809]
Yang, W.L., Yang, Y.H., Yan, X.F., Wang, X., 2008. Determination of phospholipid content in tree leaves by high performance liquid chromatography. Journal of Northeast Forestry University, 36(9):61–62 (in Chinese).
Yapa, P.A.J., Kawasaki, T., Matsumoto, H., 1986. Changes of some membrane-associated enzyme activities and degradation of membrane phospholipids in cucumber roots due to Ca2+ starvation. Plant and Cell Physiology, 27(2): 223–232.
Yuan, H.Y., Chen, L.G., Paliyath, G., Sullivan, A., Murr, D.P., 2005. Characterization of microsomal and mitochondrial phospholipase D activities and cloning of a phospholipase D alpha cDNA from strawberry fruits. Plant Physiology and Biochemistry, 43(6):535–547. [doi:10.1016/j.plaphy.2005.04.003]
Author information
Authors and Affiliations
Corresponding author
Additional information
Project (No. 30771513) supported by the National Natural Science Foundation of China
Rights and permissions
About this article
Cite this article
Zhao, Yy., Qian, Cl., Chen, Jc. et al. Responses of phospholipase D and lipoxygenase to mechanical wounding in postharvest cucumber fruits. J. Zhejiang Univ. Sci. B 11, 443–450 (2010). https://doi.org/10.1631/jzus.B0900357
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1631/jzus.B0900357