外源褪黑素降低黄瓜体内霜霉威残留的生理生化分析

doi:10.11924/j.issn.1000-6850.casb2021-1000

摘要/Abstract

摘要：

为探究褪黑素(melatonin,Mel)对黄瓜体内农药降解中的调控作用,以黄瓜‘Y3F604’为材料,以氨基甲酯类农药霜霉威(C₉H₂₀N₂O₂)为处理试剂,采用灌根的方法研究外源施用1 mmol/L Mel对黄瓜体内霜霉威残留的影响,同时探究O₂^-·、H₂O₂、MDA、GSH和GSSG含量以及抗氧化酶和解毒酶的变化情况。结果表明,外源1 mmol/L的Mel处理显著降低了霜霉威处理48 h后黄瓜体内的霜霉威残留量。在霜霉威胁迫下,黄瓜叶片中O₂^-·、H₂O₂和MDA的含量明显增加,而外源Mel处理能够有效抑制霜霉威胁迫下黄瓜体内的O₂^-·、H₂O₂和MDA积累。与对照相比,外源Mel处理显著提高了霜霉威胁迫下SOD、CAT、POD、APX、GPX等抗氧化物酶活性,进一步促进GSH含量和总谷胱甘肽含量的增加,提高GSH/GSSG、GST与GR解毒酶活性。因此,外源施用Mel可提高黄瓜在霜霉威胁迫下的抗氧化能力和解毒能力,从而加速黄瓜体内霜霉威的代谢,是促进蔬菜残留农药降解的新途径。

关键词: 黄瓜, 褪黑素, 霜霉威, 抗氧化酶, 解毒酶

Abstract:

In order to explore the regulation effect of melatonin (Mel) on degradation of residual pesticide in cucumber, cucumber variety ‘Y3F604’ was taken as test material and the carbamate pesticide propamocarb (C₉H₂₀N₂O₂) was applied to study the effect of 1 mmol/L Mel on propamocarb residues by irrigating the root. At the same time, the content of O₂^-·, H₂O₂, MDA, GSH and GSSG, and the change of antioxidant enzymes and detoxification enzymes after the application of propamocarb were investigated. The results showed that the exogenous application of 1 mmol/L Mel significantly reduced pesticide residues in cucumbers after the propamocarb treatment for 48 h. Under the stress of propamocarb, the content of O₂^-·, H₂O₂and MDA in leaves increased significantly, while exogenous Mel treatment could effectively inhibit the accumulation of O₂^-·, H₂O₂and MDA in cucumber. Compared with the control, exogenous Mel treatment could significantly increase the activity of cucumber SOD, CAT, POD, APX and GPX under the stress of propamocarb, and further increase the content of GSH and total glutathione, the ratio of GSH/GSSG, as well as the activity of GST and GR. Therefore, exogenous Mel treatment could induce the ability of antioxidant and detoxification under the stress of propamocarb, thus promoting the metabolism of propamocarb in cucumber. This study provides a new way to promote the degradation of pesticide residues in vegetables.

Key words: cucumber, melatonin, propamocarb, antioxidant enzyme, detoxification enzyme

中图分类号:

S642.2

吕薇, 李胜男, 冯国军, 杨晓旭, 刘畅, 闫志山, 刘大军. 外源褪黑素降低黄瓜体内霜霉威残留的生理生化分析[J]. 中国农学通报, 2022, 38(28): 107-113.

LV Wei, LI Shengnan, FENG Guojun, YANG Xiaoxu, LIU Chang, YAN Zhishan, LIU Dajun. Physiological and Biochemical Analysis of Exogenous Melatonin for Reducing Propamocarb Residues in Cucumber[J]. Chinese Agricultural Science Bulletin, 2022, 38(28): 107-113.

图/表 6

图1. 外源褪黑素处理对黄瓜体内霜霉威残留量的影响不同字母表示数据间差异显著(P<0.05),下同

图2. 外源褪黑素处理对农药胁迫下黄瓜体内O2-·产生速率和H2O2含量的影响

图3. 外源褪黑素处理对农药胁迫下黄瓜体MDA含量的影响

处理	还原型谷胱甘肽含量/[mmol/(g·FW)]	氧化型谷胱甘肽含量/[mmol/(g·FW)]	总谷胱甘肽/[mmol/(g·FW)]	还原型谷胱甘肽和氧化型谷胱甘肽的比值
CK	0.46c	0.38a	0.84b	1.21c
Mel	0.43cd	0.39a	0.82b	1.10c
霜霉威	0.74b	0.25b	0.99ab	2.96b
Mel+霜霉威	0.82a	0.24b	1.06a	3.42a

表1. 外源褪黑素对农药胁迫下黄瓜体内GSH-GSSG含量的影响

图4. 外源褪黑素处理对农药胁迫下黄瓜体内抗氧化酶活性的影响

图5. 外源褪黑素处理对农药胁迫下黄瓜体内解毒酶活性的影响

参考文献 44

[1]	禹化强. 大棚黄瓜主要病害的防治研究[J]. 中国果菜, 2015(2):48-52.
[2]	唐晶. 霜霉威在烟田中的残留行为及效应研究[D]. 长沙: 湖南农业大学, 2014.
[3]	WU P. Propamocarb effects on antioxidant parameters and osmolyte levels of cucumber fruit differing in propamocarb residual capacity[J]. Research journal of biotechnology, 2014,9(6):99-104.
[4]	农业部农药检定所. GB 22621—2008,霜霉威原药[S]. 北京: 中国标准出版社, 2009.
[5]	柳亚男, 李敏敏, 范蓓. 等. 超高效液相色谱串联质谱法测定霜霉威在番茄和土壤中残留及消解动态[J]. 环境化学, 2015,34(6):1072-1077.
[6]	吴鹏. 黄瓜低霜霉威残留性的生理生化及分子基础研究[D]. 哈尔滨: 东北农业大学, 2013.
[7]	彭小琴. 24-表油菜素内酯和百菌清处理对葡萄光合和生理代谢的调控[D]. 杨凌: 西北农林科技大学, 2014.
[8]	ZHOU Y, XIA X, YU G, et al. Brassinosteroids play a critical role in the regulation of pesticide metabolism in crop plants[J]. Scientific reports, 2015,5:9018. doi: 10.1038/srep09018 pmid: 25761674
[9]	YIN Y L, ZHOU Y, ZHOU Y H, et al. Interplay between mitogen-activated protein kinase and nitric oxide in brassinosteroid-induced pesticide metabolism in Solanum lycopersicum[J]. Journal of hazardous materials, 2016,316:221-231. doi: 10.1016/j.jhazmat.2016.04.070 URL
[10]	杜丽亚, 章钢娅, 靳伟. 土壤含水量和胡敏酸对有机氯农药降解的影响[J]. 土壤学报, 2006(2):332-336.
[11]	刘新宇, 陈鹏, 张光辉. 外源脯氨酸对番茄体内残留百菌清降解的调控作用[J]. 浙江农业学报, 2020,32(3):437-446. doi: 10.3969/j.issn.1004-1524.2020.03.08
[12]	李胜男. CsHMGB降低黄瓜霜霉威残留的分子机制[D]. 哈尔滨: 东北农业大学, 2020.
[13]	李娟起. 褪黑素缓解自毒物质胁迫下黄瓜种子萌发和幼苗生长的相关生理机制[D]. 北京: 中国农业大学, 2017.
[14]	YAN Y, SUN S, ZHAO N, et al. COMT1 overexpression resulting in increased melatonin biosynthesis contributes to the alleviation of carbendazim phytotoxicity and residues in tomato plants[J]. Environmental pollution, 2019,252:51-61. doi: S0269-7491(18)33812-0 pmid: 31146238
[15]	王学奎. 植物生理生化实验原理和技术[M]. 北京: 高等教育出版社, 2006:105-280.
[16]	李合生. 植物生理生化实验原理和技术[M]. 北京: 高等教育出版社, 2000:165-185.
[17]	赵世杰, 苍晶. 植物生理学实验指导[M]. 北京: 中国农业出版社, 2015:109-230.
[18]	RAI A C, SINGH M, SHAH K. Effect of water withdrawal on formation of free radical, proline accumulation and activities of antioxidant enzymes in ZAT12-transformed transgenic tomato plants[J]. Plant physiology and biochemistry, 2012,61(4):108-114. doi: 10.1016/j.plaphy.2012.09.010 URL
[19]	SHAH K, KUMAR R G, VERMA S, et al. Effect of cadmium on lipid peroxidation, superoxide anion generation and activities of antioxidant enzymes in growing rice seedlings[J]. Plant science, 2001,161(6):1135-1144. doi: 10.1016/S0168-9452(01)00517-9 URL
[20]	MA F W, CHENG L L. Exposure of the shaded side of apple fruit to full sun leads to up-regulation of both xanthoohyll cycle and the ascorbate-glutathione cycle[J]. Plant science, 2004,166(6):1479-1486. doi: 10.1016/j.plantsci.2004.01.024 URL
[21]	RAHMAN I, KODE A, BISWAS S K. Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method[J]. Nature protocols, 2006,1(6):3159-3165. pmid: 17406579
[22]	KNOIZER O C, DURNER J, BOGER P. Alteration in the antioxidative system of suspension cultivated soybean cells (Glycine max) induced by oxidative stress[J]. Physiologla plantarum, 1996,97:388-396.
[23]	KOMIVES A V, KOMIVS T, DUTKA F. Effect of thiocarbamate herbicides on the activity of glutathione S-transferase in maize[J]. Cereal research communications, 1985,13:253-257
[24]	刘芳芳. 黄瓜种质资源低农药残留性评价[D]. 哈尔滨: 东北农业大学, 2008.
[25]	LI Hui, LUO Yi-Feng, WANG Yong-Sheng, et al. Using ROS as a second messenger, NADPH oxidase 2 mediates macrophage senescence via interaction with NF-κB during Pseudomonas aeruginosa infection[J]. Oxidative medicine cellular longevity, 2018:1-10.
[26]	QI J S, SONG C P, WANG B S, et al. ROS signaling and stomatal movement in plant responses to drought stress and pathogen attack[J]. Journal of integrative plant biology, 2018(9):67-88.
[27]	GILL S S, TUTEJA N. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants[J]. Plant physiology and biochemistry, 2010,48(12):909-930. doi: 10.1016/j.plaphy.2010.08.016 pmid: 20870416
[28]	刘新宇, 陈鹏, 张光辉. 外源脯氨酸对番茄体内残留百菌清降解的调控作用[J]. 浙江农业学报, 2020,32(3):437-446. doi: 10.3969/j.issn.1004-1524.2020.03.08
[29]	刘娜. 外源褪黑素缓解黄瓜幼苗吡虫啉胁迫的生理和分子机制[D]. 兰州: 甘肃农业大学, 2021.
[30]	徐刚, 甘彩玲, 方艳琼. 等. 氮离子注入对麻疯树幼苗抗氧化能力的影响[J]. 植物生理学报, 2011,47(11):1091-1095.
[31]	徐向东. 高温胁迫下外援褪黑素对黄瓜幼苗活性氧代谢的影响[J]. 应用生态学报, 2010,21(5):1295-1300.
[32]	魏志为. 褪黑素对光逆境下苹果的保护功能研究[D]. 杨凌: 西北农林科技大学, 2019.
[33]	付晴晴, 谭雅中, 翟衡. 等. 葡萄中褪黑素对NaHCO₃胁迫的响应及外源褪黑素缓解NaHCO₃胁迫的作用机制[J]. 植物生理学报, 2017,53(12):2114-2124.
[34]	张俊康, 马丽, 吴姝青. 等. 外源褪黑素对软枣猕猴桃低温伤害的缓解效应[J]. 植物生理学报, 2020,56(5):1081-1087.
[35]	WANG P, YIN L H, LIANG D, et al. Delayed senescence of apple leaves by exogenous melatonin treatment: toward regulating the ascorbate-glutathione cycle[J]. Journal of pineal research, 2012,53(1):11-20. doi: 10.1111/j.1600-079X.2011.00966.x pmid: 21988707
[36]	LI S N, XIN M, LUAN J, et al. Overexpression of CsHMGB alleviates phytotoxicity and propamocarb residues in cucumber[J]. Frontiers in plant science, 2020,11.
[37]	MITTOVA V, THEODOULOU F L, KIDDLE G, et al. Coordinate induction of glutathione biosynthesis and glutathione- metabolizing enzymes is correlated with salt tolerance in tomato[J]. FEBS letters, 2003,554(3):417-421. doi: 10.1016/S0014-5793(03)01214-6 URL
[38]	WANG S C, LIANG D, LI C, et al. Influence of drought stress on the cellular ultra structure and antioxidant system in leaves of drought-tolerant and drought-sensitive apple root-stocks[J]. Plant physiology and biochemistry, 2012,51:81-89. doi: 10.1016/j.plaphy.2011.10.014 URL
[39]	阮海华. 外源一氧化氮对盐胁迫下小麦幼苗叶片谷胱甘肽抗氧化酶系统的影响[J]. 作物学报, 2005,31(9):1144-1149.
[40]	韩一林. 毛竹幼苗抗氧化酶和AsA-GSH循环对高温干旱及协同胁迫的响应[J]. 浙江农林大学学报, 2018,35(2):268-276.
[41]	孙舒畅. 褪黑素在番茄镉胁迫抗性中的功能研究[D]. 杭州: 浙江大学, 2020.
[42]	DIXON D P, DAVIS B G, EDWARDS R. Functional divergence in the glutathione transferase superfamily in plants. I dentification of two classes with putative functions in redox homeostasis in Arabidopsis thaliana[J]. The journal of biological chemistry, 2002,277(34):30859-30869. doi: 10.1074/jbc.M202919200 URL
[43]	MOST P, PAPENBROCK J. Possible roles of plant sulfurtransferases in detoxification of cyanide, reactive oxygen species, selected heavy metals and arsenate[J]. Molecules, 2015,20(1):1410-1423. doi: 10.3390/molecules20011410 pmid: 25594348
[44]	丁顺华, 陈姗, 卢从明. 植物叶绿体谷胱甘肽还原酶的功能研究进展[J]. 植物生理学报, 2016,52(11):1703-1709.