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
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.
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
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
Atiqur RAHMAN, Md. Hasan Sofiur RAHMAN, Md. Shakil UDDIN, Naima SULTANA, Shirin AKHTER, Ujjal Kumar NATH, Shamsun Nahar BEGUM, Md. Mazadul ISLAM, Afroz NAZNIN, Md. Nurul AMIN, Sharif AHMED, Akbar HOSAIN. Advances in DNA methylation and its role in cytoplasmic male sterility in higher plants[J]. >Journal of Integrative Agriculture, 2024, 23(1): 1-19.