Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (9): 1781-1789.doi: 10.3864/j.issn.0578-1752.2022.09.007

• PLANT PROTECTION • Previous Articles     Next Articles

The Role and Mechanism of Linoleyl Ethanolamide in Plant Resistance Against Botrytis cinerea in Tomato

SHAO ShuJun(),HU ZhangJian,SHI Kai*()   

  1. College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058
  • Received:2021-11-06 Revised:2021-12-10 Online:2022-05-01 Published:2022-05-19
  • Contact: Kai SHI E-mail:ssjun@zju.edu.cn;kaishi@zju.edu.cn

RichHTML

33

PDF

196

Abstract:

【Background】Gray mold caused by Botrytis cinerea is one of the important diseases of tomato and causes significant yield losses up to 30%-40%. Nowadays, chemical pesticide is usually used in tomato production, which is effective but increases the risk of food safety and results in environmental pollution. N-acylethanolamines (NAEs) are a kind of naturally lipid bioactive compounds in plants, which have been identified to have a variety of immune functions in mammals, however, its function and the underlying mechanism in plant immunity are still unclear.【Objective】The objective of this study is to investigate the effects of NAEs on tomato plant defense against B. cinerea infection, and to provide a basis for the development of green control technology of tomato gray mold.【Method】The B. cinerea was cultured in medium containing NAE 18:0, NAE 18:2, NAE 22:5, respectively, to evaluate their effects on B. cinerea growth. Tomato ‘Moneymaker’ plants were infected by B. cinerea with or without exogenous NAE 18:0, NAE 18:2, NAE 22:5, and disease index and fluorescence parameters of tomato leaves were measured. qRT-PCR was used to analyze the relative gene expression of B. cinerea Actin in tomato leaves that infected by B. cinerea with or without NAE 18:2 treatment. Transcript abundance of defense-related genes (e.g. PI I, PR-1, NPR1, Nr, ACO1, PYR1a), and contents of plant hormones (e.g. JA, SA, ETH, ABA, IAA) were measured. Fluorescence parameters of tomato leaves and the relative gene expression of B. cinerea Actin were analyzed in ethylene-insensitive mutant infected by B. cinerea with NAE 18:2.【Result】The growth of B. cinerea was not affected by exogenous NAEs treatment during in vitro culture. Exogenous application of NAEs could significantly improve the resistance of tomato plants to B. cinerea, and alleviate the decrease of photosystem II photochemical efficiency (ΦPSII) caused by B. cinerea infection. NAE 18:2 had the best effect on tomato plant defense against B. cinerea among the NAEs, which obviously reduced the disease index and the Actin transcript level of B. cinerea by 60%. The expression levels of PI I, PR-1, NPR1, Nr and ACO1 could be induced by B. cinerea infection but not by NAE 18:2 treament. The expression levels of PI I, Nr and ACO1 were up-regulated when plants were pre-treated by NAE 18:2 before B. cinerea infection, and the expression level of ACO1 was the highest. Compared to the control, the contents of SA, JA, IAA and ETH in the leaves were increased significantly after B. cinerea infection, while only the contents of ETH were further increased when pre-treated by NAE 18:2. Moreover, exogenous NAE 18:2 pre-treatment could not improve the defense against B. cinerea in the ETH-insensitive mutant nr.【Conclusion】Exogenous NAE18:2 treatment can increase leaf photosynthesis, transcript abundance of defense-related genes, and the content of plant hormone ETH. It induce the resistance of tomato plants to gray mold, which may depend on the ETH signaling pathway.

Key words: tomato, N-acylethanolamines (NAE), linoleyl ethanolamide (NAE 18:2), gray mold, Botrytis cinerea, ethylene (ETH)

Table 1

Specific primers designed for qRT-PCR"

基因Gene 前引物Forward primer (5′-3′) 后引物Reverse primer (5′-3′)
Actin TGGTCGGAATGGGACAGAAG CTCAGTCAGGAGAACAGGGT
B. c Actin ACTCATATGTTGGAGATGAAGCGCA AATGTTACCATACAAATCCTTACGGA
PI I GAAGTAATTAAGCAGCCACAATATG GCCCCCCTTATTTTTTCC
PR-1 ATCTCATTGTTACTCACTTGTC AACGAGCCCGACCA
NPR1 GGGAAAGATAGCAGCACG GTCCACACAAACACACACATC
PYR1a GAGAACACCGGTTGAGGAAT TGTGACTCGCATCACGACTA
Nr TTTGCGTGTCCAGGTTAGAG GGAGGCATAGGTAGCAGAGG
ETR2 CTAGTGCATCTAGGGTGCGA TCCATCCATTCCAACTCTCA
ACO1 GCGCCACTCTATTGTGGTTA TGCATCACTTCCTGGATTGT

Fig. 1

Effects of N-acylethanolamines treatment on in vitro B. cinerea hyphal growth"

Fig. 2

Effects of exogenous N-acylethanolamines on defense to tomato gray mold"

Fig. 3

Effects of exogenous NAE 18:2 on the transcript abundance of defense-related genes (A) and the endogenous hormone content (B, C) under B. cinerea infection in tomato plants"

Fig. 4

Effects of exogenous NAE 18:2 on nr tomato plant defense to gray mold"

[1] PANTHEE D R, CHEN F. Genomics of fungal disease resistance in tomato. Current Genomics, 2010, 11(1): 30-39.
doi: 10.2174/138920210790217927
[2] SINGH V K, SINGH A K, KUMAR A. Disease management of tomato through PGPB: Current trends and future perspective. 3 Biotech, 2017, 7: 255.
doi: 10.1007/s13205-017-0896-1
[3] KUEHL F, JACOB T, GANLEY O, ORMOND R, MEISINGER M. The identification of N-(2-hydroxyethyl)-palmitamide as a naturally occurring anti-inflammatory agent. Journal of the American Chemical Society, 1957, 79(20): 5577-5578.
doi: 10.1021/ja01577a066
[4] CHAPMAN K D, VENABLES B, MARKOVIC R, BLAIR R W, BETTINGER C. N-acylethanolamines in seeds. Quantification of molecular species and their degradation upon imbibition. Plant Physiology, 1999, 120(4): 1157-1164.
doi: 10.1104/pp.120.4.1157
[5] VENABLES B J, WAGGONER C A, CHAPMAN K D. N- acylethanolamines in seeds of selected legumes. Phytochemistry, 2005, 66(16): 1913-1918.
doi: 10.1016/j.phytochem.2005.06.014
[6] BLANCAFLOR E B, CHAPMAN K D. Similarities between endocannabinoid signaling in animal systems and N-acylethanolamine metabolism in plants//BALUSKA F, MANCUSO S, VOLKMANN D. Communication in Plants:Neuronal Aspects of Plant Life. Florence, Italy: Springer, 2006: 205-219.
[7] CHAPMAN K D. Occurrence, metabolism, and prospective functions of N-acylethanolamines in plants. Progress in Lipid Research, 2004, 43(4): 302-327.
doi: 10.1016/j.plipres.2004.03.002
[8] TRIPATHY S, VENABLES B, CHAPMAN K D. N-acylethanolamines in signal transduction of elicitor perception. Attenuation of alkalinization response and activation of defense gene expression. Plant Physiology, 1999, 121(4): 1299-1308.
doi: 10.1104/pp.121.4.1299
[9] BLANCAFLOR E B, HOU G, CHAPMAN K D. Elevated levels of N-lauroylethanolamine, an endogenous constituent of desiccated seeds, disrupt normal root development in Arabidopsis thaliana seedlings. Planta, 2003, 217(2): 206-217.
doi: 10.1007/s00425-003-0985-8
[10] MOTES C M, PECHTER P, YOO C M, WANG Y S, CHAPMAN K D, BLANCAFLOR E B. Differential effects of two phospholipase D inhibitors, 1-butanol and N-acylethanolamine, on in vivo cytoskeletal organization and Arabidopsis seedling growth. Protoplasma, 2005, 226(3/4): 109-123.
doi: 10.1007/s00709-005-0124-4
[11] TEASTER N D, MOTES C M, TANG Y H, WIANT W C, COTTER M Q, WANG Y S, KILARU A, VENABLES B J, HASENSTEIN K H, GONZALEZ G, BLANCAFLOR E B, CHAPMAN K D. N- acylethanolamine metabolism interacts with abscisic acid signaling in Arabidopsis thaliana seedlings. The Plant Cell, 2007, 19(8): 2454-2469.
doi: 10.1105/tpc.106.048702
[12] KIM S C, KANG L, NAGARAJ S, BLANCAFLOR E B, MYSORE K S, CHAPMAN K D. Mutations in Arabidopsis fatty acid amide hydrolase reveal that catalytic activity influences growth but not sensitivity to abscisic acid or pathogens. The Journal of Biological Chemistry, 2009, 284(49): 34065-34074.
doi: 10.1074/jbc.M109.059022
[13] ZHANG Y, GUO W M, CHEN S M, HAN L, LI Z M. The role of N-lauroylethanolamine in the regulation of senescence of cut carnations (Dianthus caryophyllus). Journal of Plant Physiology, 2007, 164(8): 993-1001.
doi: 10.1016/j.jplph.2006.07.003
[14] CHAPMAN K D, TRIPATHY S, VENABLES B, DESOUZA A D. N-acylethanolamines: Formation and molecular composition of a new class of plant lipids. Plant Physiology, 1998, 116(3): 1163-1168.
doi: 10.1104/pp.116.3.1163
[15] KANG L, WANG Y S, UPPALAPATI S R, WANG K, TANG Y, VADAPALLI V, VENABLES B J, CHAPMAN K D, BLANCAFLOR E B, MYSORE K S. Overexpression of a fatty acid amide hydrolase compromises innate immunity in Arabidopsis. The Plant Journal, 2008, 56(2): 336-349.
doi: 10.1111/j.1365-313X.2008.03603.x
[16] SHRESTHA A, SCHIKORA A. AHL-priming for enhanced resistance as a tool in sustainable agriculture. FEMS Microbiology Ecology, 2020, 96(12): fiaa226.
doi: 10.1093/femsec/fiaa226
[17] HU Z J, SHAO S J, ZHENG C F, SUN Z H, SHI J Y, YU J Q, QI Z Y, SHI K. Induction of systemic resistance in tomato against Botrytis cinerea by N-decanoyl-homoserine lactone via jasmonic acid signaling. Planta, 2018, 247(5): 1217-1227.
doi: 10.1007/s00425-018-2860-7
[18] EL OIRDI M, ABD EL RAHMAN T, RIGANO L, EL HADRAMI A, RODRIGUEZ M C, DAAYF F, VOJNOV A, BOUARAB K. Botrytis cinerea manipulates the antagonistic effects between immune pathways to promote disease development in tomato. The Plant Cell, 2011, 23(6): 2405-2421.
doi: 10.1105/tpc.111.083394
[19] 蔡银杰, 周小林, 杨献娟, 曹均尧, 冒锦富. 大棚番茄灰霉病发生的影响因子初步研究. 中国植保导刊, 2007, 27(10): 21-23.
CAI Y J, ZHOU X L, YANG X J, CAO J Y, MAO J F. A preliminary analysis on the factors affecting Botrytis cinema in green house. China Plant Protection, 2007, 27(10): 21-23. (in Chinese)
[20] SUN Y J, GENG Q W, DU Y P, YANG X H, ZHAI H. Induction of cyclic electron flow around photosystem I during heat stress in grape leaves. Plant Science, 2017, 256: 65-71.
doi: 10.1016/j.plantsci.2016.12.004
[21] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 2001, 25(4): 402-408.
doi: 10.1006/meth.2001.1262
[22] WU J Q, HETTENHAUSEN C, MELDAU S, BALDWIN I T. Herbivory rapidly activates MAPK signaling in attacked and unattacked leaf regions but not between leaves of Nicotiana attenuata. The Plant Cell, 2007, 19(3): 1096-1122.
doi: 10.1105/tpc.106.049353
[23] ZHANG S, LI X, SUN Z H, SHAO S J, HU L F, YE M, ZHOU Y H, XIA X J, YU J Q, SHI K. Antagonism between phytohormone signalling underlies the variation in disease susceptibility of tomato plants under elevated CO2. Journal of Experimental Botany, 2015, 66(7): 1951-1963.
doi: 10.1093/jxb/eru538
[24] ALGER B E. Endocannabinoids: Getting the message across. Proceedings of the National Academy of Sciences of the United States of America, 2004, 101(23): 8512-8513.
[25] KILARU A, TAMURA P, ISAAC G, WELTI R, VENABLES B J, SEIER E. CHAPMAN K D. Lipidomic analysis of N- acylphosphatidylethanolamine molecular species in Arabidopsis suggests feedback regulation by N-acylethanolamines. Planta, 2012, 236(3): 809-824.
doi: 10.1007/s00425-012-1669-z
[26] KEEREETAWEEP J, BLANCAFLOR E B, HORNUNG E, FEUSSNER I, CHAPMAN K D. Ethanolamide oxylipins of linolenic acid negatively regulates Arabidopsis seedling development. The Plant Cell, 2013, 25(10): 3824-3840.
doi: 10.1105/tpc.113.119024
[27] 杜颖, 付丹妮, 邹益泽, 白雪松, 姜震, 程攻, 纪明山, 祁之秋. 2017年辽宁省番茄灰霉菌对腐霉利的抗药性现状及机制研究. 中国蔬菜, 2018(1): 58-65.
DU Y, FU D N, ZOU Y Z, BAI X S, JIANG Z, CHENG G, JI M S, QI Z Q. Studies on drug resistance of tomato Botrytis cinerea to procymidone at Liaoning Province in 2017. China Vegetables, 2018(1): 58-65. (in Chinese)
[28] ABUQAMAR S, MOUSTAFA K, TRAN L S. Mechanisms and strategies of plant defense against Botrytis cinerea. Critical Reviews in Biotechnology, 2017, 37(2): 262-274.
doi: 10.1080/07388551.2016.1271767
[29] BROOKS D M, BENDER C L, KUNKEL B N. The Pseudomonas syringae phytotoxin coronatine promotes virulence by overcoming salicylic acid-dependent defences in Arabidopsis thaliana. Molecular Plant Pathology, 2005, 6(6): 629-639.
doi: 10.1111/j.1364-3703.2005.00311.x
[30] GRANT M, LAMB C. Systemic immunity. Current Opinion in Plant Biology, 2006, 9(4): 414-420.
doi: 10.1016/j.pbi.2006.05.013
[31] 张燕, 夏更寿, 赖志兵. 植物抗灰霉病分子机制的研究进展. 生物技术通报, 2018, 34(2): 10-24.
doi: 10.13560/j.cnki.biotech.bull.1985.2018-0040
ZHANG Y, XIA G S, LAI Z B. Rencent advances in molecular mechanisms of plant responses against Botrytis cinerea. Biotechnology Bulletin, 2018, 34(2): 10-24. (in Chinese)
doi: 10.13560/j.cnki.biotech.bull.1985.2018-0040
[32] KENDE H. Ethylene biosynthesis. Annual Review of Plant Physiology and Plant Molecular Biology, 1993, 44: 283-307.
doi: 10.1146/annurev.pp.44.060193.001435
[33] YANG S F, HOFFMAN N E. Ethylene biosynthesis and its regulation in higher plants. Annual Review of Plant Physiology, 1984, 35: 155-189.
doi: 10.1146/annurev.pp.35.060184.001103
[34] BROEKAERT W F, DELAURE S L, DE BOLLE M F C, CAMMUE B P A. The role of ethylene in host-pathogen interactions. Annual Review of Phytopathology, 2006, 44: 393-416.
doi: 10.1146/annurev.phyto.44.070505.143440
[35] TSUCHISAKA A, YU G X, JIN H L, ALONSO J M, ECKER J R, ZHANG X M, GAO S, THEOLOGIS A. A combinatorial interplay among the 1-aminocyclopropane-1-carboxylate isoforms regulates ethylene biosynthesis in Arabidopsis thaliana. Genetics, 2009, 183(3): 979-1003.
doi: 10.1534/genetics.109.107102
[36] PENNINCKX I A, THOMMA B P, BUCHALA A, METRAUX J P, BROEKAERT W F. Concomitant activation of jasmonate and ethylene response pathways is required for induction of a plant defensin gene in Arabidopsis. The Plant Cell, 1998, 10(12): 2103-2113.
doi: 10.1105/tpc.10.12.2103
[37] COHN J R, MARTIN G B. Pseudomonas syringae pv. tomato type III effectors AvrPto and AvrPtoB promote ethylene-dependent cell death in tomato. The Plant Journal, 2005, 44(1): 139-154.
doi: 10.1111/j.1365-313X.2005.02516.x
[38] BERROCAL-LOBO M, MOLINA A, SOLANO R. Constitutive expression of ETHYLENE-RESPONSE-FACTOR1 in Arabidopsis confers resistance to several necrotrophic fungi. The Plant Journal, 2002, 29(1): 23-32.
doi: 10.1046/j.1365-313x.2002.01191.x
[39] ABUQAMAR S, CHAI M F, LUO H L, SONG F M, MENGISTE T. Tomato protein kinase 1b mediates signaling of plant responses to necrotrophic fungi and insect herbivory. The Plant Cell, 2008, 20(7): 1964-1983.
doi: 10.1105/tpc.108.059477
[1] ZHANG DongMei, ZHOU XinXin, XIAO GuiLin, ZENG XiangGuo, WANG ChunYan, WANG ZeXian, HAN YongChao. Phenotypic Characteristics of Strawberry Floral Organs in Response to Botrytis cinerea Infection and Methods for Gray Mold Resistance Evaluation [J]. Scientia Agricultura Sinica, 2026, 59(7): 1456-1466.
[2] WANG YuPing, FU Zhi, SUN JiaYing, MU XiaoMeng, LIU HuiLin, GUO JinYun, SONG WenJing, HOU LeiPing, ZHAO HaiLiang. Evaluation of the Mitigating Effect and Application Efficacy of Melatonin Applied at the Seedling Stage on Short-Term Chilling Stress in Tomato Plants [J]. Scientia Agricultura Sinica, 2026, 59(7): 1523-1535.
[3] WU YuanYuan, LÜ ShuWen, ZHANG ZiJun, WANG Tao, ZHANG YiMing, BU LingChao, ZOU QingDao, JIANG Jing. Mixed Major Gene+Polygene Genetic Analysis of Blossom-End Scar Size in Tomato Fruit [J]. Scientia Agricultura Sinica, 2026, 59(5): 1060-1069.
[4] ZHANG Min, LI Xin, ZHANG Yong, ZHONG DePing, LU XiaoXiao, HE ShuMin, CHEN DongHong, LI Ye, LI RongXia, HUANG ZeJun, WANG XiaoXuan, GUO YanMei, DU YongChen, LIU HongHai, LI JunMing, LIU Lei. Genetic and Interaction Analysis of High Soluble Solid Content Loci in Processing Tomato [J]. Scientia Agricultura Sinica, 2025, 58(9): 1816-1829.
[5] ZHANG YaFeng, DONG WeiJin, LI QiYun, LU Yang, ZHANG ZhengKun, SUI Li. Effect of Interaction Between Beauveria bassiana and Potassium on Tomato Fruit Quality [J]. Scientia Agricultura Sinica, 2025, 58(6): 1131-1144.
[6] WANG ShaoHua, SHEN NianQiao, CHU TianRan, WU YongHan, LI KangNing, SHI YanXia, XIE XueWen, LI Lei, FAN TengFei, LI BaoJu, CHAI ALi. Effects of Tomato-Rice Rotation on Physicochemical Properties and Microbial Communities of Soil with Continuous Cropping Obstacles in Cangnan, Zhejiang [J]. Scientia Agricultura Sinica, 2025, 58(4): 692-703.
[7] ZHANG TianYu, LI Bai, ZANG JinPing, CAO HongZhe, DONG JinGao, XING JiHong, ZHANG Kang. Genome-Wide Identification and Expression Analysis of HMG Family Genes in Botrytis cinerea [J]. Scientia Agricultura Sinica, 2025, 58(4): 704-718.
[8] SU XiaoMei, YANG ZongHui, LIU ShuMei, ZHANG ZongJie, LÜ HongJun, HOU LiXia. Development and Application of A KASP Marker-Based Identification System for Tomato Varieties [J]. Scientia Agricultura Sinica, 2025, 58(22): 4746-4756.
[9] GUO MengZe, ZHANG Lei, SUN PingPing, JIANG Biao, YAN JinQiang, LI ZhengNan. Molecular Characterization and Evolutionary Dynamics of Tomato Leaf Curl New Delhi Virus Isolate from Wax Gourd (Benincasa hispida) in Guangdong [J]. Scientia Agricultura Sinica, 2025, 58(19): 3890-3904.
[10] MA Jia, PENG JieLi, WANG Xu, JIA Nan, LI MengKai, HU Dong. Effects of Streptomyces sp. TOR3209 on Chlorophyll Synthesis and Polyamine Content in Tomato Plants Under Low Temperature Stress [J]. Scientia Agricultura Sinica, 2025, 58(15): 3064-3080.
[11] ZHAO YuLei, XIN JiaLu, LI ChengNan, LI Shan, XIE XuFei, YIN Xiao. Screening of Target Genes Downstream of VviERF045, a Transcription Factor Associated with Gray Mold Resistance in Vitis vinifera [J]. Scientia Agricultura Sinica, 2025, 58(13): 2578-2590.
[12] LI XiaoYan, DU YaDan, HU XiaoTao, LU YiNing, GU XiaoBo. The Influence of Nitrogen Application Under Aerated Drip Irrigation on the Hydraulic Characteristics and Photosynthetic Capacity of Tomato [J]. Scientia Agricultura Sinica, 2025, 58(11): 2225-2238.
[13] SUN ZhaoAn, ZHANG YiWen, JIANG LiHua, LI ZhaoJun, GUO Xin, CAO Hui, MENG FanQiao. Effects of Tomato Grafting and Nitrogen Fertilization on Fertilizer Nitrogen Fate and Nitrogen Balance [J]. Scientia Agricultura Sinica, 2024, 57(4): 755-764.
[14] PEI ShuYao, CAO HongXia, ZHANG ZeYu, ZHAO FangYang, LI ZhiJun. Physiological Response of Potted Tomatoes to NaCl and Na2SO4 Brackish Water Irrigation [J]. Scientia Agricultura Sinica, 2024, 57(3): 570-583.
[15] MA Jia, PENG JieLi, JIA Nan, WANG Xu, WANG ZhanWu, HU Dong. Effects of Streptomyces sp. TOR3209 on Chlorophyll Fluorescence Characteristics and Xanthophyll Cycle in Tomato Plants Under Cold Stress [J]. Scientia Agricultura Sinica, 2024, 57(22): 4522-4540.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd