Please wait a minute...
Journal of Integrative Agriculture  2016, Vol. 15 Issue (2): 326-338    DOI: 10.1016/S2095-3119(15)61135-2
Physiology·Biochemistry·Cultivation·Tillage Advanced Online Publication | Current Issue | Archive | Adv Search |
Effects of abiotic stress and hormones on the expressions of five 13-CmLOXs and enzyme activity in oriental melon (Cucumis melo var. makuwa Makino)
 LIU Jie-ying, ZHANG Chong, SHAO Qi, TANG Yu-fan, CAO Song-xiao, GUO Xiao-ou, JIN Ya-zhong, QI Hong-yan
1、Key Laboratory of Protected Horticulture of Liaoning Province/Key Laboratory of Protected Horticulture, Ministry of Education/
College of Horticulture, Shenyang Agricultural University, Shenyang 110866, P.R.China
2、College of Agriculture, Heilongjiang Bayi Agricultural University, Daqing 163319, P.R.China
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
摘要  Lipoxygenases (LOXs) are a group of non-heme, iron-containing enzymes and extensively involved in plant growth and development, ripening and senescence, stress responses, biosynthesis of regulatory molecules and defense reaction. In our previous study, 18 LOXs in melon genome were screened and identified, and five 13-LOX genes (CmLOX08, CmLOX10, CmLOX12, CmLOX13 and CmLOX18) were predicted to locate in chloroplast. Phylogenetic analysis result showed that the five genes have high homology with jasmonic acid (JA) biosynthesis-related LOXs from other plants. In addition, promoter analysis revealed that motifs of the five genes participate in gene expression regulated by hormones and stresses. Therefore, we analyzed the expressions of the five genes and LOX activity in leaves of four-leaf stage seedlings of oriental melon cultivar Yumeiren under abiotic stress: wounding, cold, high temperature and hydrogen peroxide (H2O2), and signal molecule treatments: methyl jasmonate (MeJA), abscisic acid (ABA) and salicylic acid (SA). Real time qPCR revealed that wounding and H2O2 induced the expressions of all the five genes. Only CmLOX08 was induced by cold while only CmLOX13 was suppressed by high temperature. ABA induced the expressions of CmLOX10 and CmLOX12 while inhibited CmLOX13 and CmLOX18. MeJA increased the 3 genes expressions except CmLOX08 and CmLOX13, whereas SA decreased the effect, apart from CmLOX12. All the abiotic stresses and signal molecules treatments increased the LOX activity in leaves of oriental melon. In summary, the results suggest that the five genes have diverse functions in abiotic stress and hormone responses, and might participate in defense response. The data generated in this study will be helpful in subcellular localization and transgenic experiment to understand their precise roles in plant defense response.

Abstract  Lipoxygenases (LOXs) are a group of non-heme, iron-containing enzymes and extensively involved in plant growth and development, ripening and senescence, stress responses, biosynthesis of regulatory molecules and defense reaction. In our previous study, 18 LOXs in melon genome were screened and identified, and five 13-LOX genes (CmLOX08, CmLOX10, CmLOX12, CmLOX13 and CmLOX18) were predicted to locate in chloroplast. Phylogenetic analysis result showed that the five genes have high homology with jasmonic acid (JA) biosynthesis-related LOXs from other plants. In addition, promoter analysis revealed that motifs of the five genes participate in gene expression regulated by hormones and stresses. Therefore, we analyzed the expressions of the five genes and LOX activity in leaves of four-leaf stage seedlings of oriental melon cultivar Yumeiren under abiotic stress: wounding, cold, high temperature and hydrogen peroxide (H2O2), and signal molecule treatments: methyl jasmonate (MeJA), abscisic acid (ABA) and salicylic acid (SA). Real time qPCR revealed that wounding and H2O2 induced the expressions of all the five genes. Only CmLOX08 was induced by cold while only CmLOX13 was suppressed by high temperature. ABA induced the expressions of CmLOX10 and CmLOX12 while inhibited CmLOX13 and CmLOX18. MeJA increased the 3 genes expressions except CmLOX08 and CmLOX13, whereas SA decreased the effect, apart from CmLOX12. All the abiotic stresses and signal molecules treatments increased the LOX activity in leaves of oriental melon. In summary, the results suggest that the five genes have diverse functions in abiotic stress and hormone responses, and might participate in defense response. The data generated in this study will be helpful in subcellular localization and transgenic experiment to understand their precise roles in plant defense response.
Keywords:  oriental melons       lipoxygenase       abiotic stress       signal molecules       gene expression  
Received: 05 January 2015   Accepted:
Fund: 

This study was financially supported by the National Natural Science Foundation of China (31272154).

Corresponding Authors:  QI Hong-yan, Tel: +86-24-88342305, E-mail: hyqiaaa@126.com,syauhongyan@hotmail.com     E-mail:  hyqiaaa@126.com,syauhongyan@hotmail.com
About author:  LIU Jie-ying, E-mail: 943832507@qq.com;* These authors contributed equally to this work.

Cite this article: 

LIU Jie-ying, ZHANG Chong, SHAO Qi, TANG Yu-fan, CAO Song-xiao, GUO Xiao-ou, JIN Ya-zhong, QI Hong-yan. 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). Journal of Integrative Agriculture, 15(2): 326-338.

Andreou A, Feussner I. 2009. Lipoxygenases-structure andreaction mechanism. Phytochemistry, 70, 1504-1510

Axelrod B. 1981. Lipoxygenase from soybeans. Methods inEnzymology, 71, 441-451

Bate N J, Rothstein S J. 1998. C6-volatiles derived from thelipoxygenase pathway induce a subset of defense-relatedgenes. The Plant Journal, 16, 561-569

Bell E, Creelman R A, Mullet J E.1995. A chloroplastlipoxygenase is required for wound-induced jasmonic acidaccumulation in Arabidopsis. Proceedings of the NationalAcademy of Sciences of the United States of America, 92,8675-8679

Bhardwaj P K, Kaur J, Sobti R C, Ahuja P S, Kumar S.2011. Lipoxygenase in Caragana jubata responds to lowtemperature, abscisic acid, methyl jasmonate and salicylicacid. Gene, 483, 49-53

Bradford M M. 1976. A rapid and sensitive method for thequantitation of micro-gram quantities of protein utilizing theprinciple of protein dye binding. Analytical Biochemistry,72, 248-254

Browse J. 2005. Jasmonate: An oxylipin signal with many rolesin plants. Vitamins and Hormones, 72, 431-456

Cho K, Kim Y C, Woo J C, Rakwal R, Agrawal G K, Yoeun S, HanO. 2012. Transgenic expression of dual positional maizelipoxygenase-1 leads to the regulation of defense-relatedsignaling molecules and activation of the antioxidativeenzyme system in rice. Plant Science, 185, 238-245

Christensen S A, Nemchenko A, Borrego E, Murray I, Sobhy IS, Bosak L, DeBlasio S, Erb M, Robert C A M, Vaughn K A,Herrfurth C, Tumlinson J, Feussner I, Jackson D, TurlingsT C J, Engelberth J, Nansen C, Meeley R, Kolomiets MV. 2013. The maize lipoxygenase, ZmLOX10, mediatesgreen leaf volatile, jasmonate and herbivore-induced plantvolatile production for defense against insect attack. ThePlant Journal, 74, 59-73

Cipollini D, Enright S, Traw M B, Bergelson J. 2004.Salicylic acid inhibits jasmonic acid-induced resistanceof Arabidopsis thaliana to Spodoptera exigua. MolecularEcology, 13, 1643-1653

Demmig-Adams B, Cohu C M, Amiard V, Zadelhoff G, VeldinkG A, Muller O, Adams W W. 2013. Emerging trade-offs- impact of photoprotectants (PsbS, xanthophylls, andvitamin E) on oxylipins as regulators of development anddefense. New Phytologist, 197, 720-729

Dong X. 1998. SA, JA, ethylene, and disease resistance inplants. Current Opinion in Plant Biology, 1, 316-323

Engelberth J, Alborn H T, Schmelz E A, Tumlinson J H. 2004.Airborne signals prime plants against insect herbivoreattack. Proceedings of the National Academy of Sciencesof the United States of America, 101, 1781-1785

Engelberth J, Seidl-Adams I, Schultz J C, Tumlinson JH. 2007. Insect elicitors and exposure to green leafyvolatiles differentially upregulate major octadecanoids andtranscripts of 12-oxo phytodienoic acid reductases in Zeamays. Molecular Plant-Microbe Interactions, 20, 707-716

Feussner I, Wasternack C. 2002. The lipoxygenase pathway.Annual Review of Plant Biology, 53, 275-297

Feys B J, Parker J E. 2000. Interplay of signaling pathways inplant disease resistance. Trends in Genetics, 16, 449-455

Garcia-Marcos A, Pacheco R, Manzano A, Aguilar E, TenlladoF. 2013. Oxylipin biosynthesis genes positively regulateprogrammed cell death during compatible infections with thesynergistic pair potato Virus X-Potato Virus Y and TomatoSpotted Wilt Virus. Journal of Virology, 87, 5769-5783

Glazebrook J. 2005. Contrasting mechanisms of defenseagainst biotrophic and necrotrophic pathogens. AnnualReviews of Phytopathology, 43, 205-227

Hall T A. 1999. BioEdit: A user-friendly biological sequencealignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95-98

Halitschke R, Baldwin I T. 2003. Antisense LOX expressionincreases herbivore performance by decreasing defenseresponse and inhibiting growth-related transcriptionalreorganization in Nicotiana attenuata. The Plant Journal,36, 794-807

Halitchke R, Ziegler J, Keinänen M, Baldwin I T. 2004. Silencingof hydroperoxide lyase and allene oxide synthase revealssubstrate and defense signaling crosstalk in NicotianaAttenuata. The Plant Journal, 40, 35-46

Heitz T, Bergey D R, Ryan C A. 1997. A gene encoding achloroplast targeted lipoxygenase in tomato leaves istransiently induced by wounding, systemin, and methyljasmonate. Plant Physiology, 114, 1085-1093

Hu T Z, Zeng H, Hu Z L, Qv X X, Chen G P. 2013. Overexpressionof the tomato 13-lipoxygenase gene Tomloxd increasesgeneration of endogenous jasmonic acid and resistance tocladosporium fulvum and high temperature. Plant MolecularBiology Reporter, 31, 1141-1149

Hwang I S, Hwang B K. 2010. The pepper 9-lipoxygenase geneCaLOX1 functionsin defense and cell death responses tomicrobial pathogens. Plant Physiology, 152, 948-967

Jia Q L, Gong Z H, Li D W. 2012. Cloning and expressioncharacterization of chloroplast-targeted 13-lipoxygenasegene (CaLOX2) in Capsicum annuum L. Journal ofAgricultural Biotechnology, 20, 126-134 (in Chinese)

Liavonchanka A, Feussner I. 2006. Lipoxygenases: Occurrence,functions and catalysis. Journal of Plant Physiology, 163,348-357

Melan M, Dong X, Endara M E, Davis K R, Ausuble F M,Peterman T K. 1993. An Arabidopsis thaliana lipoxygenasegene can be induced by pathogens, abscisic acid, andmethyl jasmonate. Plant Physiology, 101, 441-450

Montillet J L, Chamnongpol S, Rustérucci C, Dat J, van deCotte B, Agnel J P, Battesti C, Inzé D, Breusegem F V,Triantaphylidès C. 2005. Fatty acid hydroperoxides andH2O2 in the execution of hypersensitive cell death in tobaccoleaves. Plant Physiology, 138, 1516-1526

Mur L A J, Kenton P, Atzorn R, Miersch O, Wasternack C.2006. The outcomes of concentration-specific interactionsbetween salicylate and jasmonate signaling include synergy, antagonism and the activation of cell death. PlantPhysiology, 140, 249-262

Nemchenko A, Kunze S, Feussner I, Kolomiets M. 2006.Duplicate maize 13-lipoxygenase genes are differentiallyregulated by circadian rhythm, cold stress, wounding,pathogen infection, and hormonal treatments. Journal ofExperimental Botany, 57, 3767-3779

Page R D M. 1996. TreeView: An application to displayphylogenetic trees on personal computers. ComputerApplications in the Biosciences, 12, 357-358

Pena-Cortes H, Albtrecht T, Prat S, Weiler W W, Willmitzer L.1993. Aspirin prevents wound-induced gene expressionin tomato leaves by blocking jasmonic acid biosynthesis.Planta, 191,123-128

Porta H, Figueroa-Balderas R E, Rocha-Sosa M. 2008.Wounding and pathogen infection induce a chloroplasttargetedlipoxygenase in the common bean (Phaseolusvulgaris L.). Planta, 227, 363-373

Royo J, Vancanneyt G, Perez A G, Sanz C, Stormann K,Rosahl S, Sanchez-Serrano J J. 1996. Characterizationof three potato lipoxygenases with distinct enzymaticactivities and different organspecific and wound-regulatedexpression patterns. The Journal of Biological Chemistry,271, 21012-21019

Ryals J, Lawton K A, Delaney T P, Friedrich L, Kessmann H,Neuenschwander U, Uknes S, Vernooij B, Weymann K.1995. Signal transduction in systemic acquired resistance.Proceedings of the National Academy of Sciences of theUnited States of America, 92, 4202-4205

Salzman R A, Brady J A, Finlayson S A, Buchanan C D,Summer E J, Sun F. 2005. Transcriptional profiling ofsorghum induced by methyl jasmonate, salicylic acid, andaminocyclopropane carboxylic acid reveals cooperativeregulation and novel gene responses. Plant Physiology,138, 352-368

Tamura K, Dudley J, Nei M, Kumar S. 2007. MEGA4: Molecularevolutionarygenetics analysis (MEGA) software version 4 0.Molecular Biology and Evolution, 24, 1596-1599

Traw M B, Kim J, Enright S, Cipollini D F, Bergelson J. 2003.Negative cross-talk between salicylate- and jasmonatemediatedpathways in the Wassilewskija ecotype ofArabidopsis thaliana. Molecular Ecology, 12, 1125-1135

Tuteja N. 2007. Abscisic acid and abiotic stress signaling. PlantSignaling & Behavior, 2, 135-138

Vicente J, Cascón T, Vicedo B, García-Agustín P, HambergM, Castresana C. 2012. Role of 9-lipoxygenase andα-dioxygenase oxylipin pathways as modulators of local andsystemic defense. Molecular Plant, 5, 914-928

Wasternack C. 2007. Jasmonates: An update on biosynthesis,signal transduction and action in plant stress response,growth and development. Annals of Botany, 100, 681-697

Wasternack C, Hause B. 2013. Jasmonates: Biosynthesis,perception, signal transduction and action in plant stressresponse, growth and development. An update to the2007 review in Annals of Botany. Annals of Botany, 111,1021-1058

Weichert H, Stenzel I, Berndt E, Wasternack C, FeussnerI. 1999. Metabolic profiling of oxylipins upon salicylatetreatment in barley leaves: Preferential induction of thereductase pathway by salicylate. FEBS Letters, 464,133-137

Yan L H, Zhai Q Z, Wei J N, Li S Y, Wang B, Huang T T, DuM M, Sun J, Kang L, Li C N, Li C Y. 2013. Role of tomatolipoxygenase D in wound-induced jasmonate biosynthesisand plant immunity to insect herbivores. PLoS Genetics,9, e1003964.

Yang X Y, Jiang W J, Yu H J. 2012. The expression profilingof the lipoxygenase (LOX) family genes during fruitdevelopment, abiotic stress and hormonal treatments incucumber (Cucumis sativus L.). International Journal ofMolecular Sciences, 13, 2481-2500

Zhang C, Jin Y Z, Liu J Y, Tang Y F, Cao S X, Qi H Y. 2014.The phylogeny and expression profiles of the lipoxygenase(LOX) family genes in the melon (Cucumis melo L.) genome.Scientia Horticulturae, 170, 94-102
[1] ZHAO Shu-ping, DENG Kang-ming, ZHU Ya-mei, JIANG Tao, WU Peng, FENG Kai, LI Liang-jun.

Optimization of slow-release fertilizer application improves lotus rhizome quality by affecting the physicochemical properties of starch [J]. >Journal of Integrative Agriculture, 2023, 22(4): 1045-1057.

[2] ZHANG Yan-mei, AO De, LEI Kai-wen, XI Lin, Jerry W SPEARS, SHI Hai-tao, HUANG Yan-ling, YANG Fa-long. Dietary copper supplementation modulates performance and lipid metabolism in meat goat kids[J]. >Journal of Integrative Agriculture, 2023, 22(1): 214-221.
[3] JIANG Yong, MA Xin-yan, XIE Ming, ZHOU Zheng-kui, TANG Jing, CHANG Guo-bin, CHEN Guo-hong, HOU Shui-sheng. Dietary threonine deficiency affects expression of genes involved in lipid metabolism in adipose tissues of Pekin ducks in a genotype-dependent manner[J]. >Journal of Integrative Agriculture, 2022, 21(9): 2691-2699.
[4] RONG Hao, YANG Wen-jing, XIE Tao, WANG Yue, WANG Xia-qin, JIANG Jin-jin, WANG You-ping. Transcriptional profiling between yellow- and black-seeded Brassica napus reveals molecular modulations on flavonoid and fatty acid content[J]. >Journal of Integrative Agriculture, 2022, 21(8): 2211-2226.
[5] LI Zhi-qi, Xie Qian, YAN Jia-hui, CHEN Jian-qing, CHEN Qing-xi. Genome-wide identification and characterization of the abiotic-stress-responsive lipoxygenase gene family in diploid woodland strawberry (Fragaria vesca)[J]. >Journal of Integrative Agriculture, 2022, 21(7): 1982-1996.
[6] AN Feng, ZHANG Kang, ZHANG Ling-kui, LI Xing, CHEN Shu-min, WANG Hua-sen, CHENG Feng. Genome-wide identification, evolutionary selection, and genetic variation of DNA methylation-related genes in Brassica rapa and Brassica oleracea[J]. >Journal of Integrative Agriculture, 2022, 21(6): 1620-1632.
[7] FAN Xiao-xue, BIAN Zhong-hua, SONG Bo, XU Hai. Transcriptome analysis reveals the differential regulatory effects of red and blue light on nitrate metabolism in pakchoi (Brassica campestris L.)[J]. >Journal of Integrative Agriculture, 2022, 21(4): 1015-1027.
[8] LIU Cong, LI De-xiong, HUANG Xian-biao, Zhang Fu-qiong, Xie Zong-zhou, Zhang Hong-yan, Liu Ji-hong. Manual thinning increases fruit size and sugar content of Citrus reticulata Blanco and affects hormone synthesis and sugar transporter activity[J]. >Journal of Integrative Agriculture, 2022, 21(3): 725-735.
[9] DU Qiao-li, FANG Yuan-peng, JIANG Jun-mei, CHEN Mei-qing, LI Xiang-yang, XIE Xin. Genome-wide identification and characterization of the JAZ gene family and its expression patterns under various abiotic stresses in Sorghum bicolor[J]. >Journal of Integrative Agriculture, 2022, 21(12): 3540-3555.
[10] DUAN Yao-ke, HAN Rong, SU Yan, WANG Ai-ying, LI Shuang, SUN Hao, GONG Hai-jun. Transcriptional search to identify and assess reference genes for expression analysis in Solanum lycopersicum under stress and hormone treatment conditions[J]. >Journal of Integrative Agriculture, 2022, 21(11): 3216-3229.
[11] LI Sheng-lan, TAN Ting-ting, FAN Yuan-fang, Muhammad Ali RAZA, WANG Zhong-lin, WANG Bei-bei, ZHANG Jia-wei, TAN Xian-ming, CHEN Ping, Iram SHAFIQ, YANG Wen-yu, YANG Feng. Response of leaf stomatal and mesophyll conductance to abiotic stress factors[J]. >Journal of Integrative Agriculture, 2022, 21(10): 2787-2804.
[12] Kashif NOOR, Hafiza Masooma Naseer CHEEMA, Asif Ali KHAN, Rao Sohail Ahmad KHAN. Expression profiling of transgenes (Cry1Ac and Cry2A) in cotton genotypes under different genetic backgrounds[J]. >Journal of Integrative Agriculture, 2022, 21(10): 2818-2832.
[13] WANG Pei-pei, WANG Zhao-ke, GUAN Le, Muhammad Salman HAIDER, Maazullah NASIM, YUAN Yong-bing, LIU Geng-sen, LENG Xiang-peng. Versatile physiological functions of the Nudix hydrolase family in berry development and stress response in grapevine[J]. >Journal of Integrative Agriculture, 2022, 21(1): 91-112.
[14] GUO Bing-bing, LI Jia-ming, LIU Xing, QIAO Xin, Musana Rwalinda FABRICE, WANG Peng, ZHANG Shao-ling, WU Ju-you. Identification and expression analysis of the PbrMLO gene family in pear, and functional verification of PbrMLO23[J]. >Journal of Integrative Agriculture, 2021, 20(9): 2410-2423.
[15] SHI Bei-bei, WANG Juan, GAO Hai-feng, ZHANG Xiao-juan, WANG Yang, MA Qing. The TaFIM1 gene mediates wheat resistance against Puccinia striiformis f. sp. tritici and responds to abiotic stress[J]. >Journal of Integrative Agriculture, 2021, 20(7): 1849-1857.
No Suggested Reading articles found!