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Journal of Zhejiang University-SCIENCE B

, Volume 19, Issue 8, pp 596–609 | Cite as

An oriental melon 9-lipoxygenase gene CmLOX09 response to stresses, hormones, and signal substances

  • Li-jun Ju
  • Chong Zhang
  • Jing-jing Liao
  • Yue-peng Li
  • Hong-yan Qi
Article

Abstract

In plants, lipoxygenases (LOXs) play a crucial role in biotic and abiotic stresses. In our previous study, five 13-LOX genes of oriental melon were regulated by abiotic stress but it is unclear whether the 9-LOX is involved in biotic and abiotic stresses. The promoter analysis revealed that CmLOX09 (type of 9-LOX) has hormone elements, signal substances, and stress elements. We analyzed the expression of CmLOX09 and its downstream genes—CmHPL and CmAOS—in the leaves of four-leaf stage seedlings of the oriental melon cultivar “Yumeiren” under wound, hormone, and signal substances. CmLOX09, CmHPL, and CmAOS were all induced by wounding. CmLOX09 was induced by auxin (indole acetic acid, IAA) and gibberellins (GA3); however, CmHPL and CmAOS showed differential responses to IAA and GA3. CmLOX09, CmHPL, and CmAOS were all induced by hydrogen peroxide (H2O2) and methyl jasmonate (MeJA), while being inhibited by abscisic acid (ABA) and salicylic acid (SA). CmLOX09, CmHPL, and CmAOS were all induced by the powdery mildew pathogen Podosphaera xanthii. The content of 2-hexynol and 2-hexenal in leaves after MeJA treatment was significantly higher than that in the control. After infection with P. xanthii, the diseased leaves of the oriental melon were divided into four levels—levels 1, 2, 3, and 4. The content of jasmonic acid (JA) in the leaves of levels 1 and 3 was significantly higher than that in the level 0 leaves. In summary, the results suggested that CmLOX09 might play a positive role in the response to MeJA through the hydroperoxide lyase (HPL) pathway to produce C6 alcohols and aldehydes, and in the response to P. xanthii through the allene oxide synthase (AOS) pathway to form JA.

Key words

9-Lipoxygenase (9-LOX) Hydroperoxide lyase (HPL) Allene oxide synthase (AOS) Green leaf volatile Jasmonic acid 

薄皮甜瓜9-脂氧合酶(9-LOX)类型的CmLOX09对逆境、激素和信号类物质的响应

中文概要

目的

研究CmLOX09及其下游基因CmHPLCmAOS 对逆境、激素和信号类物质的响应,进一步测定茉莉酸甲酯(MeJA)处理后叶片中绿叶挥发物的含量以及接种白粉病菌后叶片中茉莉酸含量的逆境、激素和信号类物质的响应,进一步测定 茉莉酸甲酯(MeJA)处理后叶片中绿叶挥发物的含量以及接种白粉病菌后叶片中茉莉酸含量的变化,探讨脂氧合酶(LOX)响应这两种胁迫的可能途径

创新点

通过对CmLOX09 启动子中顺式作用原件的分析预测,首次研究薄皮甜瓜9-LOX类型的CmLOX09 对机械损伤、激素、信号类物质以及生物胁迫的响应

方法

利用Plant CARE 软件对CmLOX09 启动子响应元 件进行预测分析(图S1);利用荧光定量聚合酶 链反应(qRT-PCR)技术分析甜瓜在机械损伤、激素、信号类物质以及生物胁迫处理后叶片中 CmLOX09CmHPLCmAOS 的表达模式;利用 气相色谱-质谱连用仪(GC-MS)测定叶片中绿 叶挥发物的含量(图4);利用高效液相色谱- 串联质谱法(HPLC-MS/MS)分析和测定叶片中 茉莉酸的含量(图6)。

结论

本研究结果显示:CmLOX09 参与机械损伤、激 素、信号类物质及白粉病菌的防御反应(图1~3, 5)。9-LOX 类型的CmLOX09 可能通过氢过氧化 物裂解酶(HPL)途径产生的绿叶挥发物(GLV) 来响应MeJA(图4),并通过丙二烯合酶(AOS) 途径产生的茉莉酸来响应真菌胁迫(图6)。综 上所述,9-LOX 类型的CmLOX09 可能在生物和 非生物胁迫反应中起重要作用。

关键词

9-脂氧合酶 氢过氧化物裂解酶 丙二烯合酶 绿叶挥发物 茉莉酸 

CLC number

S652.2 

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Supplementary material

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An oriental melon 9-lipoxygenase gene CmLOX09 response to stresses, hormones, and signal substances

References

  1. Alam MM, Nahar K, Hasanuzzaman M, et al., 2014. Exogenous jasmonic acid modulates the physiology, antioxidant defense and glyoxalase systems in imparting drought stress tolerance in different Brassica species. Plant Biotechnol Rep, 8(3):279–293. https://doi.org/10.1007/s11816-014-0321-8 CrossRefGoogle Scholar
  2. Ameye M, Audenaert K, de Zutter N, et al., 2015. Priming of wheat with the green leaf volatile Z-3-hexenyl acetate enhances defense against Fusarium graminearum but boosts deoxynivalenol production. Plant Physiol, 167(4):1671–1684. https://doi.org/10.1104/pp.15.00107 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Andreou A, Feussner I, 2009. Lipoxygenases—structure and reaction mechanism. Phytochemistry, 70(13-14):1504–1510. https://doi.org/10.1016/j.phytochem.2009.05.008 CrossRefPubMedGoogle Scholar
  4. Bae KS, Rahimi S, Kim YJ, et al., 2016. Molecular characterization of lipoxygenase genes and their expression analysis against biotic and abiotic stresses in Panax ginseng. Eur J Plant Pathol, 145(2):331–343. https://doi.org/10.1007/s10658-015-0847-9 CrossRefGoogle Scholar
  5. Bell E, Creelman RA, Mullet JE, 1995. A chloroplast lipoxygenase is required for wound-induced jasmonic acid accumulation in Arabidopsis. Proc Natl Acad Sci USA, 92(19):8675–8679. https://doi.org/10.1073/pnas.92.19.8675 CrossRefPubMedGoogle Scholar
  6. Bhardwaj PK, Kaur J, Sobti RC, et al., 2011. Lipoxygenase in Caragana jubata responds to low temperature, abscisic acid, methyl jasmonate and salicylic acid. Gene, 483(1-2):49–53. https://doi.org/10.1016/j.gene.2011.05.014 CrossRefPubMedGoogle Scholar
  7. Birkett MA,Campbell CAM, Chamberlain K, et al., 2000. New roles for cis-jasmone as an insect semiochemical and in plant defense. Proc Natl Acad Sci USA, 97(16):9329–9334. https://doi.org/10.1073/pnas.160241697 CrossRefPubMedGoogle Scholar
  8. Brash AR, 1999. Lipoxygenases: occurrence, functions, catalysis, and acquisition of substrate. J Biol Chem, 274(34):23679–23682. https://doi.org/10.1074/jbc.274.34.23679 CrossRefPubMedGoogle Scholar
  9. Chauvin A, Lenglet A, Wolfender JL, et al., 2016. Paired hierarchical organization of 13-lipoxygenases in Arabidopsis. Plants, 5(2):16. https://doi.org/10.3390/plants5020016 CrossRefPubMedCentralGoogle Scholar
  10. Christensen SA, Kolomiets MV, 2011. The lipid language of plant–fungal interactions. Fungal Genet Biol, 48(1):4–14. https://doi.org/10.1016/j.fgb.2010.05.005 CrossRefPubMedGoogle Scholar
  11. Christensen SA, Nemchenko A, Borrego E, et al., 2013. The maize lipoxygenase, ZmLOX10, mediates green leaf volatile, jasmonate and herbivore-induced plant volatile production for defense against insect attack. Plant J, 74(1):59–73. https://doi.org/10.1111/tpj.12101 CrossRefPubMedGoogle Scholar
  12. Ding H, Lai JB, Wu Q, et al., 2016. Jasmonate complements the function of Arabidopsis lipoxygenase3 in salinity stress response. Plant Sci, 244:1–7. https://doi.org/10.1016/j.plantsci.2015.11.009 CrossRefPubMedGoogle Scholar
  13. Durrant WE, Dong X, 2004. Systemic acquired resistance. Annu Rev Phytopathol, 42:185–209. https://doi.org/10.1146/annurev.phyto.42.040803.140421 CrossRefPubMedGoogle Scholar
  14. Feussner I, Wasternack C, 2002. The lipoxygenase pathway. Annu Rev Plant Biol, 53:275–297. https://doi.org/10.1146/annurev.arplant.53.100301.135248 CrossRefPubMedGoogle Scholar
  15. Gao XQ, Stumpe M, Feussner I, et al., 2008. A novel plastidial lipoxygenase of maize (Zea mays) ZmLOX6 encodes for a fatty acid hydroperoxide lyase and is uniquely regulated by phytohormones and pathogen infection. Planta, 227(2):491–503. https://doi.org/10.1007/s00425-007-0634-8 CrossRefPubMedGoogle Scholar
  16. Griffiths A, Barry C, Alpuche-Solis AG, et al., 1999. Ethylene and developmental signals regulate expression of lipoxygenase genes during tomato fruit ripening. J Exp Bot, 50(335):793–798. https://doi.org/10.1093/jxb/50.335.793 CrossRefGoogle Scholar
  17. Guo XM, Stotz HU, 2007. Defense against Sclerotinia sclerotiorum in Arabidopsis is dependent on jasmonic acid, salicylic acid, and ethylene signaling. Mol Plant Microbe Interact, 20(11):1384–1395. https://doi.org/10.1094/MPMI-20-11-1384 CrossRefPubMedGoogle Scholar
  18. Halitschke R, Baldwin LT, 2003. Antisense LOX expression increases herbivore performance by decreasing defense responses and inhibiting growth-related transcriptional reorganization in Nicotiana attenuata. Plant J, 36(6):794–807. https://doi.org/10.1046/j.1365-313X.2003.01921.x CrossRefPubMedGoogle Scholar
  19. Heitz T, Bergey DR, Ryan CA, 1997. A gene encoding a chloroplast-targeted lipoxygenase in tomato leaves is transiently induced by wounding, systemin, and methyl jasmonate. Plant Physiol, 114(3):1085–1093. https://doi.org/10.1104/pp.114.3.1085 CrossRefPubMedPubMedCentralGoogle Scholar
  20. Hou YL, Meng K, Han Y, et al., 2015. The persimmon 9-lipoxygenase gene DkLOX3 plays positive roles in both promoting senescence and enhancing tolerance to abiotic stress. Front Plant Sci, 6:1073. https://doi.org/10.3389/fpls.2015.01073 CrossRefPubMedPubMedCentralGoogle Scholar
  21. Hou YL, Bai QY, Meng K, et al., 2018. Overexpression of persimmon 9-lipoxygenase DkLOX3 confers resistance to Pseudomonas syringae pv. tomato DC3000 and Botrytis cinerea in Arabidopsis. Plant Growth Regul, 84(1):179–189. https://doi.org/10.1007/s10725-017-0331-y CrossRefGoogle Scholar
  22. Hu TZ, Zeng H, Hu ZL, et al., 2013. Overexpression of the tomato 13-lipoxygenase gene TomloxD increases generation of endogenous jasmonic acid and resistance to Cladosporium fulvum and high temperature. Plant Mol Biol Rep, 31(5):1141–1149. https://doi.org/10.1007/s11105-013-0581-4 CrossRefGoogle Scholar
  23. Hu TZ, Hu ZL, Zeng H, et al., 2015. Tomato lipoxygenase D involved in the biosynthesis of jasmonic acid and tolerance to abiotic and biotic stress in tomato. Plant Biotechnol Rep, 9(1):37–45. https://doi.org/10.1007/s11816-015-0341-z CrossRefGoogle Scholar
  24. Hwang IS, Hwang BK, 2010. The pepper 9-lipoxygenase gene CaLOX1 functions in defense and cell death responses to microbial pathogens. Plant Physiol, 152(2):948–967. https://doi.org/10.1104/pp.109.147827 CrossRefPubMedPubMedCentralGoogle Scholar
  25. Ivanov I, Heydeck D, Hofheinz K, et al., 2010. Molecular enzymology of lipoxygenases. Arch Biochem Biophys, 503(2):161–174. https://doi.org/10.1016/j.abb.2010.08.016 CrossRefPubMedGoogle Scholar
  26. Kallenbach M, Gilardoni PA, Allmann S, et al., 2011. C12 derivatives of the hydroperoxide lyase pathway are produced by product recycling through lipoxygenase-2 in Nicotiana attenuata leaves. New Phytol, 191(4):1054–1068. https://doi.org/10.1111/j.1469-8137.2011.03767.x CrossRefPubMedGoogle Scholar
  27. Keereetaweep J, Blancaflor EB, Hornung E, et al., 2015. Lipoxygenase-derived 9-hydro(pero)xides of linoleoylethanolamide interact with ABA signaling to arrest root development during Arabidopsis seedling establishment. Plant J, 82(2):315–327. https://doi.org/10.1111/tpj.12821 CrossRefPubMedGoogle Scholar
  28. la Camera S, Balagué C, Göbel C, et al., 2009. The Arabidopsis Patatin-like protein 2 (PLP2) plays an essential role in cell death execution and differentially affects biosynthesis of oxylipins and resistance to pathogens. Mol Plant Microbe Interact, 22(4):469–481. https://doi.org/10.1094/MPMI-22-4-0469 CrossRefPubMedGoogle Scholar
  29. León J, Royo J, Vancanneyt G, et al., 2002. Lipoxygenase H1 gene silencing reveals a specific role in supplying fatty acid hydroperoxides for aliphatic aldehyde production. J Biol Chem, 277(1):416–423. https://doi.org/10.1074/jbc.M107763200 CrossRefPubMedGoogle Scholar
  30. Liavonchanka A, Feussner I, 2006. Lipoxygenases: occurrence, functions and catalysis. Plant Physiol, 163(3):348–357. https://doi.org/10.1016/j.jplph.2005.11.006 CrossRefGoogle Scholar
  31. Lim CW, Han SW, Hwang IS, et al., 2015. The pepper lipoxygenase CaLOX1 plays a role in osmotic, drought and high salinity stress response. Plant Cell Physiol, 56(5):930–942. https://doi.org/10.1093/pcp/pcv020 CrossRefPubMedGoogle Scholar
  32. Liu JY, Zhang C, Shao Q, et al., 2016. Effects of abiotic stress and hormones on the expressions of five 13-CmLOXs and enzyme activity in oriental melon (Cucumis melo var. makuwa Makino). J Integr Agric, 15(2):326–338. https://doi.org/10.1016/S2095-3119(15)61135-2 CrossRefGoogle Scholar
  33. Liu WN, Hildebrand DF, Grayburn WS, et al., 1991. Effects of exogenous auxins on expression of lipoxygenases in cultured soybean embryos. Plant Physiol, 97(3):969–976. https://doi.org/10.1104/pp.97.3.969 CrossRefPubMedPubMedCentralGoogle Scholar
  34. Lv JY, Rao JP, Zhu YM, et al., 2014. Cloning and expression of lipoxygenase genes and enzyme activity in ripening persimmon fruit in response to GA and ABA treatments. Postharvest Biol Technol, 92:54–61. https://doi.org/10.1016/j.postharvbio.2014.01.015 CrossRefGoogle Scholar
  35. Maccarrone M, van Zadelhoff G, Veldink GA, et al., 2000. Early activation of lipoxygenase in lentil (Lens culinaris) root protoplasts by oxidative stress induces programmed cell death. Eur J Biochem, 267(16):5078–5084. https://doi.org/10.1046/j.1432-1327.2000.01564.x CrossRefPubMedGoogle Scholar
  36. Marmey P, Jalloul A, Alhamdia M, et al., 2007. The 9-lipoxygenase GhLOX1 gene is associated with the hypersensitive reaction of cotton Gossypium hirsutum to Xanthomonas campestris pv malvacearum. Plant Physiol Biochem, 45(8):596–606. https://doi.org/10.1016/j.plaphy.2007.05.002 CrossRefPubMedGoogle Scholar
  37. Maschietto V, Marocco A, Malachova A, et al., 2015. Resistance to Fusarium verticillioides and fumonisin accumulation in maize inbred lines involves an earlier and enhanced expression of lipoxygenase (LOX) genes. J Plant Physiol, 188:9–18. https://doi.org/10.1016/j.jplph.2015.09.003 CrossRefPubMedGoogle Scholar
  38. Mostofa MG, Hossain MA, Fujita M, 2015. Trehalose pretreatment induces salt tolerance in rice (Oryza sativa L.) seedlings: oxidative damage and co-induction of antioxidant defense and glyoxalase systems. Protoplasma, 252(2):461–475. https://doi.org/10.1007/s00709-014-0691-3 CrossRefPubMedGoogle Scholar
  39. Neill SJ, Desikan R, Clarke A, et al., 2002. Hydrogen peroxide and nitric oxide as signalling molecules in plants. J Exp Bot, 53(372):1237–1247. https://doi.org/10.1093/jexbot/53.372.1237 CrossRefPubMedGoogle Scholar
  40. Nemchenko A, Kunze S, Feussner I, et al., 2006. Duplicate maize 13-lipoxygenase genes are differentially regulated by circadian rhythm, cold stress, wounding, pathogen infection, and hormonal treatments. J Exp Bot, 57(14):3767–3779. https://doi.org/10.1093/jxb/erl137 CrossRefPubMedGoogle Scholar
  41. Park JH, Halitschke R, Kim HB, et al., 2002. A knock-out mutation in allene oxide synthase results in male sterility and defective wound signal transduction in Arabidopsis due to a block in jasmonic acid biosynthesis. Plant J, 31(1):1–12. https://doi.org/10.1046/j.1365-313X.2002.01328.x CrossRefPubMedGoogle Scholar
  42. Porta H, Figueroa-Balderas RE, Rocha-Sosa M, 2008. Wounding and pathogen infection induce a chloroplasttargeted lipoxygenase in the common bean (Phaseolus vulgaris L.). Planta, 227(2):363–373. https://doi.org/10.1007/s00425-007-0623-y CrossRefPubMedGoogle Scholar
  43. Rancé I, Fournier J, Esquerré-Tugayé MT, 1998. The incompatible interaction between Phytophthora parasitica var. nicotianae race 0 and tobacco is suppressed in transgenic plants expressing antisense lipoxygenase sequences. Proc Natl Acad Sci USA, 95(11):6554–6559. https://doi.org/10.1073/pnas.95.11.6554 CrossRefPubMedGoogle Scholar
  44. Sayegh-Alhamdia M, Marmey P, Jalloul A, et al., 2008. Association of lipoxygenase response with resistance of various cotton genotypes to the bacterial blight disease. J Phytopathol, 156(9):542–549. https://doi.org/10.1111/j.1439-0434.2008.01409.x CrossRefGoogle Scholar
  45. Shen JY, Tieman D, Jones JB, et al., 2014. A 13-lipoxygenase, TomloxC, is essential for synthesis of C5 flavour volatiles in tomato. J Exp Bot, 65(2):419–428. https://doi.org/10.1093/jxb/ert382 CrossRefPubMedPubMedCentralGoogle Scholar
  46. Tang YF, Zhang C, Cao SX, et al., 2015. The effect of CmLOXs on the production of volatile organic compounds in four aroma types of melon (Cucumis melo). PLoS ONE, 10(11):e0143567. https://doi.org/10.1371/journal.pone.0143567 CrossRefPubMedPubMedCentralGoogle Scholar
  47. Wang R, Shen WB, Liu LL, et al., 2008. A novel lipoxygenase gene from developing rice seeds confers dual position specificity and responds to wounding and insect attack. Plant Mol Biol, 66(4):401–414. https://doi.org/10.1007/s11103-007-9278-0 CrossRefPubMedGoogle Scholar
  48. Xin ZJ, Zhang LP, Zhang ZQ, et al., 2014. A tea hydroperoxide lyase gene, CsiHPL1, regulates tomato defense response against Prodenia litura (Fabricius) and Alternaria alternata f. sp. lycopersici by modulating green leaf volatiles (GLVs) release and jasmonic acid (JA) gene expression. Plant Mol Biol Rep, 32(1):62–69. https://doi.org/10.1007/s11105-013-0599-7 CrossRefGoogle Scholar
  49. Yan LH, Zhai QZ, Wei JN, et al., 2013. Role of tomato lipoxygenase D in wound-induced jasmonate biosynthesis and plant immunity to insect herbivores. PLoS Genet, 9(12):e1003964. https://doi.org/10.1371/journal.pgen.1003964 CrossRefPubMedPubMedCentralGoogle Scholar
  50. Yang XY, Jiang WJ, Yu HJ, 2012. The expression profiling of the lipoxygenase (LOX) family genes during fruit development, abiotic stress and hormonal treatments in cucumber (Cucumis sativus L.). Int J Mol Sci, 13(2):2481–2500. https://doi.org/10.3390/ijms13022481 CrossRefPubMedPubMedCentralGoogle Scholar
  51. Zhang C, Jin Y, Liu JY, et al., 2014. The phylogeny and expression profiles of the lipoxygenase (LOX) family genes in the melon (Cucumis melo L.) genome. Scientia Horticulturae, 170:94–102. https://doi.org/10.1016/j.scienta.2014.03.005 CrossRefGoogle Scholar
  52. Zhang C, Shao Q, Cao SX, et al., 2015. Effects of postharvest treatments on expression of three lipoxygenase genes in oriental melon (Cucumis melo var. makuwa Makino). Postharvest Biol Technol, 110:229–238. https://doi.org/10.1016/j.postharvbio.2015.08.024 CrossRefGoogle Scholar
  53. Zhang JH, Jia WS, Yang JC, et al., 2006. Role of ABA in integrating plant responses to drought and salt stresses. Field Crop Res, 97(1):111–119. https://doi.org/10.1016/j.fcr.2005.08.018 CrossRefGoogle Scholar
  54. Zhou GX, Qi JF, Ren N, et al., 2009. Silencing OsHI-LOX makes rice more susceptible to chewing herbivores, but enhances resistance to a phloem feeder. Plant J, 60(4):638–648. https://doi.org/10.1111/j.1365-313X.2009.03988.x CrossRefPubMedGoogle Scholar

Copyright information

© Zhejiang University and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Key Laboratory of Protected Horticulture of Education Ministry and Liaoning Province, College of HorticultureShenyang Agricultural UniversityShenyangChina

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