Exogenous melatonin mediates the antioxidant system and phenylpropanoid metabolism to induce resistance to leaf spot disease in oat

doi:10.11686/cyxb2023426

Abstract

Abstract:

The aim of this work was to determine how exogenous melatonin affects the resistance of oat （Avena sativa） to leaf spot disease caused by the pathogen Drechslera avenae. Seedlings of the oat cultivar ‘Molasses’ were subjected to foliar spraying with melatonin （0， 10， 100， 250， 500， and 1000 μmol·L^-1） followed by inoculation with the pathogen. At 9 days after inoculation， the disease index was calculated， and physiological indicators and the transcript levels of genes related to the antioxidant system and phenylpropanoid metabolism were determined. The results showed that leaf spot disease significantly inhibited oat plant growth， but the disease index was significantly decreased when melatonin was applied. The degree to which the disease index was decreased differed depending on the melatonin concentration. The in vitro induction efficiency of the pathogen， the activities of catalase and phenylalanine ammonia lyase， and the transcript level of the gene encoding ascorbate peroxidase were highest in the 100 μmol·L^-1 melatonin treatment， and were 50.72%， 80.32%， 3.88%， and 148.00% higher than their respective values in the control. The lowest superoxide anion production rate and malondialdehyde content were in the 250 μmol·L^-1 melatonin treatment， and were 28.27% and 86.62% lower than their respective values in the control. Compared with the control， in the 250 μmol·L^-1 melatonin treatment， the activities of superoxide dismutase， peroxidase， ascorbate peroxidase， cinnamic acid-4-hydroxylase， and 4-coumaric acid∶CoA ligase were increased by 128.23%， 137.14%， 74.00%， 52.65%， and 26.73%， respectively； the contents of lignin， total phenols， and flavonoids were increased by 1.40%， 10.33%， and 13.70%， respectively， and the transcript levels of genes encoding superoxide dismutase， peroxidase， catalase， phenylalanine ammonia lyase， 4-coumaric acid∶CoA ligase， and cinnamic acid-4-hydroxylase were increased by 352.56%， 80.32%， 65.71%， 26.65%， 52.65%， and 20.11%， respectively. Seedlings treated with 500 μmol·L^-1 melatonin showed the lowest hydrogen peroxide content （75.45% lower than that in the control） and the highest chlorophyll content （81.35% higher than that in the control）. According to the above results， the best concentration of melatonin for inducing resistance to leaf spot in oat plants is 250 μmol·L^-1. The results of this study demonstrate a new approach for the control of leaf spot disease in oat.

Key words: oat, melatonin, induced disease resistance, leaf spot

Zheng WANG, Xin LI, Jian-gui ZHANG, Ji-kuan CHAI, Gui-qin ZHAO, Kui-ju NIU. Exogenous melatonin mediates the antioxidant system and phenylpropanoid metabolism to induce resistance to leaf spot disease in oat[J]. Acta Prataculturae Sinica, 2024, 33(10): 135-146.

Figures/Tables 10

Table 1 The qRT-PCR primer design

引物Primer	上游引物Upstream primer	下游引物Downstream primer
超氧化物歧化酶SOD	TGTCGATCGGGCAGATGATG	CCCCACCACCAGGTTTCATT
过氧化物酶POD	AACGTGCAGAACCCGATCAT	AGCGGTAGTCCCTCTGATGT
过氧化氢酶CAT	CGGAGTCCCTCCACATGTTC	TTGACGTAGTGGGACTTGCC
抗坏血酸过氧化物酶APX	TTGTCGCTCTATCTGGTGGC	AGCCCCTCAGATTCTCCCTT
苯丙氨酸解氨酶PAL	CTATCGATCTGCGCCACCTT	TGTGATTGGTGCTCAGGGTC
4-香豆酸∶辅酶A连接酶4CL	AGGATGCTGCTGTCGTATCG	TGCGACGAACTGCTTGATCT
肉桂酸-4-羟化酶C4H	AACTACGGCGACTTCATCCC	TCCTTGAAGAGCTTGAGGCG
肌动蛋白基因The actin gene	GAGCGGGAAATTGTAAGGGAC	ATGGATGGCTGGAAGAGGAC

Fig.1 The effect of exogenous melatonin on the growth of pathogenic bacteria

Fig.2 The effect of exogenous melatonin on the growth of pathogenic bacteria

Fig.3 Effect of exogenous melatonin induced resistance to leaf spot disease in oat seedlings

Fig.4 The effect of exogenous melatonin on the disease index and chlorophyll content of oat leaves after inoculation with pathogenic bacteria during the seedling stage

Fig.5 Effects of exogenous melatonin on H2O2 content， superoxide anion production rate， and malondialdehyde content in leaves of oat seedlings inoculated with pathogenic bacteria

Fig.6 Effect of exogenous melatonin on antioxidant enzyme activity in oat seedlings after inoculation with pathogenic bacteria

Fig.7 Effect of exogenous melatonin on the activity of phenylpropanoid metabolism related enzymes in oat leaves after inoculation with pathogenic bacteria during seedling stage

Fig.8 The effect of exogenous melatonin on the lignin， total phenol， and flavonoid content of oat leaves after inoculation with pathogenic bacteria during the seedling stage

Fig.9 The effect of exogenous melatonin on the relative expression of enzyme genes in oat leaves after inoculation with pathogenic bacteria during the seedling stage

References 41

1	Ye X L， Gan Z， Wan Y， et al. Advances and perspectives in forage oat breeding. Acta Prataculturae Sinica， 2023， 32（2）： 160-177.
	叶雪玲，甘圳，万燕，等. 饲用燕麦育种研究进展与展望. 草业学报， 2023， 32（2）： 160-177.
2	Zhang M， Zhang M L， Guo J， et al. Summary of distribution， production， nutritional and physiological value of Avena sativa L. in China. Inner Mongolia Agricultural Science and Technology， 2014（2）： 116-118， 126.
	张曼，张美莉，郭军，等. 中国燕麦分布、生产及营养价值与生理功能概述. 内蒙古农业科技， 2014（2）： 116-118， 126.
3	Nie X M， Zhao G Q， Sun H Y， et al. Incidence of oat leaf spot and pathogen identification in the main oat production areas of Gansu Province. Acta Prataculturae Sinica， 2020， 29（4）： 157-167.
	聂秀美，赵桂琴，孙浩洋，等. 甘肃省燕麦主产区叶斑病调查及病原鉴定. 草业学报， 2020， 29（4）： 157-167.
4	Dong B Z， Zhang X Y， Zhao G Q， et al. Evaluation of pesistance of oat varieties to leaf spot pathogens//Proceedings of the 2015 Academic Annual Meeting of the Chinese Society for Plant Pathology. Beijing： China Agriculture Press， 2015： 246.
	东保柱，张笑宇，赵桂琴，等. 燕麦品种对叶斑病菌的抗性评价//中国植物病理学会2015年学术年会论文集. 北京：中国农业出版社， 2015： 246.
5	Dong B Z， Zhang X Y， Zhao G Q， et al. Indoor toxicity and field control effect of different fungicides on oat leaf spot disease// Proceedings of the 2015 Academic Annual Meeting of the Chinese Society of Plant Pathology. Beijing： China Agriculture Press， 2015： 247.
	东保柱，张笑宇，赵桂琴，等. 不同杀菌剂对燕麦叶斑病的室内毒力和田间防效//中国植物病理学会2015年学术年会论文集. 北京：中国农业出版社， 2015： 247.
6	Song J L， Cheng X K， Liu P F， et al. Current situation of multidrug resistance of plant pathogens to fungicides. Plant Protection， 2021， 47（6）： 28-33， 65.
	宋佳露，程星凯，刘鹏飞，等. 植物病原菌对杀菌剂多药抗性的发生现状. 植物保护， 2021， 47（6）： 28-33， 65.
7	Dong L L. Study on plant induced disease resistance mechanism. Value Engineering， 2010， 29（24）： 233.
	董玲玲. 关于植物诱导抗病机理的研究. 价值工程， 2010， 29（24）： 233.
8	Afreen F， Zobayed S M A， Kozai T. Melatonin in Glycyrrhiza uralensis： response of plant roots to spectral quality of light and UV-B radiation. Journal of Pineal Research， 2006， 41（2）： 108-115.
9	Chen X， Latordap P， Liu F Q. Exogenous melatonin enhances rice plant resistance against Xanthomonas oryzae pv. oryzae. Plant Disease， 2020， 104（6）： 1701-1708.
10	Li C， He Q L， Zhang F， et al. Melatonin enhances cotton immunity to Verticillium wilt via manipulating lignin and gossypol biosynthesis. The Plant Journal， 2019， 100（1）： 784-800.
11	Liu Y P. Physiological mechanism of exogenous melation inducing resistance to clubroot in Chinese cabbage. Yangling： Northwest A&F University， 2022.
	刘亚萍. 外源褪黑素对大白菜根肿病抗性的生理机制研究. 杨凌：西北农林科技大学， 2022.
12	Yuan J H， Cao L X， Shi B H， et al. Evaluation of main oat cultivars or lines resistance against Drechslera leaf spot in northwest Hebei Province. China Plant Protection， 2014， 34（2）： 31-34.
	袁军海，曹丽霞，石碧红，等. 冀西北地区燕麦主栽品种（系）对叶斑病抗性鉴定. 中国植保导刊， 2014， 34（2）： 31-34.
13	Arnon D I. Copper enzymes in isolated chloroplasts. Plant Physiology， 1949， 24（1）： 1-15.
14	Chakrabarty D， Datta S K. Micropropagation of gerbera： Lipid peroxidation and antioxidant enzyme activities during acclimatization process. Acta Physiologiae Plantarum， 2008， 30（1）： 325-331.
15	Ke D， Sun G， Wang Z. Effects of superoxide radicals on ACC synthase activity in chilling-stressed etiolated mungbean seedlings. Plant Growth Regulation， 2007， 51（1）： 83-91.
16	Kumar G， Knowles N R. Changes in lipid peroxidation and lipolytic and free-radical scavenging enzyme activities during aging and sprouting of potato （Solanum tuberosum） seed-tubers. Plant Physiology， 1993， 102（1）： 115-124.
17	Beauchamp C， Fridovich I. Superoxide dismutase： Improved assays and an assay applicable to acrylamide gels. Analytical Biochemistry， 1971， 44（1）： 276-287.
18	Munoz-Munoza J L， Garcia-Molina F， Garcia-Ruiz P A， et al. Enzymatic and chemical oxidation of trihydroxylated phenols. Food Chemistry， 2009， 113（1）： 435-444.
19	Li L， Li N H， Jiang S M. Experimental guide of plant physiology module. Beijing： Science Press， 2009： 37-103.
	李玲，李娘辉，蒋素梅. 植物生理学模块实验指导. 北京：科学出版社， 2009： 37-103.
20	Nakano Y， Asada K. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant and Cell Physiology， 1980， 22（1）： 867-880.
21	Cao J K， Jiang W B， Zhao Y M. Guidance on postharvest physiological experiments of fruits and vegetables. Beijing： China Light Industry Press， 2017.
	曹建康，姜微波，赵玉梅. 果蔬采后生理生化实验指导. 北京：中国轻工出版社， 2017.
22	Xu J， Zhang Z， Li X， et al. Effect of nitrous oxide against Botrytis cinerea and phenylpropanoid pathway metabolism in table grapes. Scientia Horticulturae， 2019， 254： 99-105.
23	Bhaskara R M V， Arul J， Angers P， et al. Chitosan treatment of wheat seeds induces resistance to Fusarium graminearum and improves seed quality. Journal of Agricultural and Food Chemistry， 1999， 47（1）： 1208-1216.
24	He J. Screening of inducing antibiotics and preliminary study on the mechanism of Taizi ginseng against leaf spot disease. Guiyang： Guizhou University， 2022.
	何洁. 太子参抗叶斑病的诱抗剂筛选及机理初探. 贵阳：贵州大学， 2022.
25	Li S E. Exogenous melatonin induces disease resistance against pathogen infection in cherry tomato fruit during postharvest storage. Hangzhou： Zhejiang Gongshang University， 2022.
	李生娥. 外源褪黑素诱导采后番茄果实抗病机制的研究. 杭州：浙江工商大学， 2022.
26	Deitz K J， Mittler R， Noctor G. Recent progress in understanding the role of reactive oxygen species in plant cell signaling. Plant Physiology， 2016， 171（3）： 1535.
27	Zhu J C， Yang Y， Lou H， et al. Regulation of Fusarium wilt resistance in cotton by exogenous melatonin. Biotechnology Bulletin， 2023， 39（1）： 243-252.
	朱金成，杨洋，娄慧，等. 外源褪黑素调控棉花枯萎病抗性研究. 生物技术通报， 2023， 39（1）： 243-252.
28	Li Z Z， Zhang S J， Xue J X， et al. Exogenous melatonin treatment induces disease resistance against Botrytis cinerea on post-harvest grapes by activating defence responses. Foods， 2022， 11（1）： 11-17.
29	Lin Y， Fan L， Xia X， et al. Melatonin decreases resistance to postharvest green mold on citrus fruit by scavenging defense-related reactive oxygen species. Postharvest Biology and Technology， 2019， 153（1）： 21-30.
30	Apel K， Hirt H. Reactive oxygen species： Metabolism， oxidative stress， and signal transduction. Annual Review of Plant Biology， 2004， 55（2）： 373-399.
31	Liang X， Lu F P， Lu H， et al. Preliminary study of the function of catalase in cassava resistance to mite. Chinese Journal of Tropical Crops， 2017， 38（2）： 343-348.
	梁晓，卢芙萍，卢辉，等. 保护酶CAT在木薯种质抗螨中的功能初步研究. 热带作物学报， 2017， 38（2）： 343-348.
32	Guo Z G， Wang M X， Cui L， et al. Research progress on the underlying mechanisms of plant defense enzymes in response to pest stress. Chinese Journal of Applied Ecology， 2018， 29（12）： 4248-4258.
	郭祖国，王梦馨，崔林，等. 6种防御酶调控植物体应答虫害胁迫机制的研究进展. 应用生态学报， 2018， 29（12）： 4248-4258.
33	Yin L H. Mechanism and resistance induced on marssonina apple blotch. Yangling： Northwest A&F University， 2014.
	殷丽华. 苹果属资源对苹果褐斑病的抗性机理及抗性诱导研究. 杨凌：西北农林科技大学， 2014.
34	Li C X， Chen L L， Zhao R R， et al. Melatonin induces disease resistance to Botrytis cinerea in tomato fruit by activating jasmonic acid signaling pathway. Journal of Agricultural and Food Chemistry， 2019， 67（1）： 6116-6124.
35	Sun Y K， Liu Z Y， Lan G P， et al. Effect of exogenous melatonin on resistance of cucumber to downy mildew. Scientia Horticulturae， 2019， 255（1）： 231-241.
36	Shang J， Wu W Z， Ma Y G. Phenylpropanoid metabolism pathway in plants. Chinese Journal of Biochemistry and Molecular Biology， 2022， 38（11）： 1467-1476.
	尚军，吴旺泽，马永贵. 植物苯丙烷代谢途径. 中国生物化学与分子生物学报， 2022， 38（11）： 1467-1476.
37	Xie F， Hao L， Wang Z J， et al. Advances in understanding how branches of the phenylpropanoid pathway respond to biotic stress. China Plant Protection， 2023， 43（2）： 23-30.
	谢凤，郝乐，王振杰，等. 苯丙烷代谢途径分支对生物胁迫响应的研究进展. 中国植保导刊， 2023， 43（2）： 23-30.
38	Li S， Xu Y， Bi Y， et al. Melatonin treatment inhibits gray mold and induces disease resistance in cherry tomato fruit during postharvest. Postharvest Biology and Technology， 2019， 157（1）： 110962.
39	Qu G， Wu W， Ba L， et al. Melatonin enhances the postharvest disease resistance of blueberries fruit by modulating the jasmonic acid signaling pathway and phenylpropanoid metabolites. Frontiers in Chemistry， 2022， 10（1）： 957581.
40	Zhang Z， Nie Y T， Cui K L， et al. Research progress on the function of melatonin in regulating growth， development and stress resistance in herbaceous species. Acta Agrestia Sinica， 2023， 31（9）： 2571-2581.
	张昭，聂宇婷，崔凯伦，等. 褪黑素调控草类植物生长发育及抗逆性功能研究进展. 草地学报， 2023， 31（9）： 2571-2581.
41	Xing X， Li L L， Yang H M， et al. Toxicity and control effects of fungicides in oat leaf spot by Drechslera avenae. Journal of Northeast Agricultural Sciences， 2023， 48（4）： 62-66.
	邢星，李乐乐，杨海明，等. 不同杀菌剂对燕麦叶斑病菌的毒力和田间防效. 东北农业科学， 2023， 48（4）： 62-66.

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