Inhibitory Effect of Resveratrol on Walnut Bacterial Black Spot Pathogen

doi:10.13560/j.cnki.biotech.bull.1985.2020-1267

Abstract

Abstract:

It aims to provide theoretical basis for the development of natural bactericide for walnut bacterial black spot,and to lay a foundation for the application of resveratrol in the prevention and control of agricultural and forestry crop diseases. The minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of resveratrol against Xanthomonas arboricola pv. juglandis(Xaj)were determined. The following experiments were carried out under the sub-inhibitory concentration of resveratrol:1)The formation of Xaj biofilm was detected by crystal violet staining. 2)The production of exopolysaccharide(EPS)was determined. 3)The secretion of extracellular enzyme was analyzed on the corresponding solid medium by biochemical method. 4)The motility of Xaj was detected on swarming medium. The results revealed that the MIC of resveratrol against Xaj was 20 μg/mL,and MBC was 45 μg/mL. The formation of Xaj biofilm was effectively inhibited by resveratrol at a sub-inhibitory concentration of 15 μg/mL. Under this concentration,the production of EPS was also significantly inhibited. Resveratrol effectively inhibited the secretion of Xaj protease,promoted the secretion of amylase,but had no effect on the secretion of cellulase. There was no effect on motility(swarming). Resveratrol has an inhibitory effect on walnut bacterial black spot pathogen,affecting the virulence factors of this bacterium,and is expected to develop a suitable plant-derived fungicide for the prevention and treatment of walnut black spot disease.

Key words: resveratrol, walnut bacterial black spot pathogen, antibacterial effect, botanical fungicides

LI Feng, CHEN Wen-wen, DENG Jiang-li, MAO Qing-li, QUAN Wen-li, MAO Ya-hui. Inhibitory Effect of Resveratrol on Walnut Bacterial Black Spot Pathogen[J]. Biotechnology Bulletin, 2021, 37(6): 58-65.

Figures/Tables 7

Fig. 1 Detection of the antibacterial activity of resveratrol against Xaj A: The plate of filter paper for qualitative analysis, No.1 was the filter paper with DMSO, No.2, 3 and 4 were the filter papers with resveratrol. B: MIC and MBC values were determined by concentration gradient dilution method. The t test was used for significance analysis, NS indicates no significant difference (P>0.05), **P<0.01, ***P<0.001, The same below

Fig. 2 Effect of resveratrol on Xaj biofilm formation A: Biofilm formation on the surface of finger tube. B: Histogram of biofilm yield

Fig. 3 Effect of resveratrol on extracellular polysaccharide production by Xaj

Fig. 4 Effect of resveratrol on extracellular protease secreted by Xaj A: Test plate containing 1% skim milk. B: Histogram of extracellular protease yield

Fig. 5 Effect of resveratrol on extracellular amylase secreted by Xaj A: Test plate containing 0.1% soluble starch. B: Histogram of extracellular amylase yield

Fig. 6 Effect of resveratrol on extracellular cellulase secreted by Xaj A: Test plate containing 0.5% carboxymethyl cellulose. B: Histogram of extracellular cellulase yield

Fig.7 Effect of resveratrol on Xaj motility A: Test plate of motility. B: Histogram of colony diameter statistics

References 23

[1]	李晓菲, 徐政. 植物源杀菌剂研究进展[J]. 南方农业, 2018, 12(13):40-42, 45.
	Li XF, Xu Z. Research progress of plant-derived fungicides[J]. Southern China Agriculture, 2018, 12(13):40-42, 45.
[2]	左卫平, 蔡昱, 刘文英, 等. 虎杖化学成分研究[J]. 中国药学杂志, 2020, 55(3):189-193.
	Zuo WP, Cai Y, Liu WY, et al. Chemical constituents of Polygonum cuspidatum[J]. Chinese Pharmaceutical Journal, 2020, 55(3):189-193.
[3]	梁明辉. 中药虎杖的研究进展[J]. 中国医药指南, 2019, 17(10):47, 54.
	Liang MH. Research progress of Polygonum cuspidatum[J]. Chinese Medicine Guide, 2019, 17(10):47, 54.
[4]	Ma DS, Tan LT, Chan K, et al. Resveratrol—potential antibacterial agent against foodborne pathogens[J]. Frontiers in Pharmacology, 2018, 9:102. doi: 10.3389/fphar.2018.00102 URL
[5]	Xia N, Daiber A, Forstermann U, et al. Antioxidant effects of resveratrol in the cardiovascular system[J]. British Journal of Pharmacology, 2017, 174(12):1633-1646. doi: 10.1111/bph.v174.12 URL
[6]	Chikara S, Nagaprashantha LD, Singhal J, et al. Oxidative stress and dietary phytochemicals:Role in cancer chemoprevention and treatment.[J]. Cancer Letters, 2018:122-134.
[7]	Zhao L, Zhang Q, Ma W, et al. A combination of quercetin and resveratrol reduces obesity in high-fat diet-fed rats by modulation of gut microbiota[J]. Food & Function, 2017, 8(12):4644-4656.
[8]	Bradamante S, Barenghi L, Villa A, et al. Cardiovascular protective effects of resveratrol[J]. Cardiovascular Drug Reviews, 2006, 22(3):169-188. doi: 10.1111/cdr.2004.22.issue-3 URL
[9]	Matsumura N, Zordoky BN, Robertson IM, et al. Co-administration of resveratrol with doxorubicin in young mice attenuates detrimental late-occurring cardiovascular changes[J]. Cardiovascular Research, 2018, 114(10):1350-1359. doi: 10.1093/cvr/cvy064 URL
[10]	Gambini J, Ingles M, Olaso G, et al. Properties of resveratrol:in vitro and in vivo studies about metabolism, bioavailability, and biological effects in animal models and humans[J]. Oxidative Medicine and Cellular Longevity, 2015: 837042.
[11]	吴翠霞, 王金信, 侯珍, 等. 白藜芦醇及其衍生物对植物病原真菌的抑菌活性[J]. 农药学学报, 2012, 14(3):283-290.
	Wu CX, Wang JX, Hou Z, et al. Inhibitory activity of resveratrol and its derivatives against phytopathogenic fungi[J]. Journal of Pesticide Science, 2012, 14(3):283-290.
[12]	李永军, 李巍伟, 王鑫, 等. 白藜芦醇对葡萄球菌抗菌活性研究[J]. 中国实验诊断学杂志, 2008(1):58-60.
	Li YJ, Li WW, Wang X, et al. Study on the antibacterial activity of resveratrol against Staphylococcus[J]. Chinese Journal of Laboratory Diagnostics, 2008(1):58-60.
[13]	苏军华, 郭勇, 袁宁璐, 等. 白藜芦醇抗病毒抗菌作用的研究现状[J]. 中国美容医学, 2012, 21(11):45-46.
	Su JH, Guo Y, Yuan NL, et al. Research status of resveratrol’s antiviral and antibacterial effects[J]. Chinese Journal of Aesthetic Medicine, 2012, 21(11):45-46.
[14]	吴翠霞. 白藜芦醇在水果贮存中的应用[J]. 世界农药, 2010, 32(2):29-30.
	Wu CX. Application of resveratrol in fruit storage[J]. World Pesticides, 2010, 32(2):29-30.
[15]	Scotton M, Bortolin E, Fiorin A, et al. Environmental and pathogenic factors inducing brown apical necrosis on fruit of English(Persian)walnut[J]. Phytopathology, 2015, 105:1427. doi: 10.1094/PHYTO-01-15-0029-R URL
[16]	Jiang S, Han S, He D, et al. Evaluating Walnut(Juglans spp. )for resistance to walnut blight and comparisons between artificial inoculation assays and field studies[J]. Australasian Plant Pathology, 2019, 48(3):221-231. doi: 10.1007/s13313-019-0621-0 URL
[17]	惠军涛, 杨峰, 杨桦, 等. 核桃黑斑病综合防治措施[J]. 西北园艺, 2016(5):36-37.
	Hui JT, Yang F, Yang H, et al. Comprehensive control measures of walnut black spot[J]. Northwest Horticulture, 2016(5):36-37.
[18]	Rigano LA, Siciliano F, Enrique R, et al. Biofilm formation, epiphytic fitness, and canker development in Xanthomonas axonopodis pv. citri[J]. Molecular Plant-Microbe Interactions, 2007, 20(10):1222, 1230. doi: 10.1094/MPMI-20-10-1222 URL
[19]	冯洁, 徐进, 许景升, 等. Ⅱ型分泌系统与植物病原细菌致病性的关系[J]. 农业生物技术学报, 2007, 15(3):365-370.
	Feng J, Xu J, Xu JS, et al. Relationship between the typeII secretion system and pathogenicity of plant pathogenic bacteria[J]. Journal of Agricultural Biotechnology, 2007, 15(3):365-370.
[20]	李峰, 刘婕, 毛雅慧. 三种植物病原菌毒力因子的初步研究[J]. 湖北工程学院学报, 2019, 39(3):32-37.
	Li F, Liu J, Mao YH. A preliminary study on the virulence factors of three plant pathogens[J]. Journal of Hubei Engineering University, 2019, 39(3):32-37.
[21]	Wallace J, Bowlin NO, Mills DM, et al. Discovery of bacterial fatty acid synthase type II inhibitors using a novel cellular bioluminescent reporter assay[J]. Antimicrobial Agents and Chemotherapy, 2015, 59(9):5775-5787. doi: 10.1128/AAC.00686-15 URL
[22]	张瑞, 李永军, 王鑫. 白藜芦醇抗菌作用的研究进展[J]. 河北医科大学学报, 2010(9):1151-1154.
	Zhang R, Li YJ, Wang X. Research progress on the antibacterial effect of resveratrol[J]. Journal of Hebei Medical University, 2010, 31(9):1151-1154.
[23]	He YW, Boon C, Zhou L, et al. Co-regulation of Xanthomonas campestris virulence by quorum sensing and a novel two-component regulatory system RavS/RavR.[J]. Molecular Microbiology, 2009, 71(6):1464-1476. doi: 10.1111/mmi.2009.71.issue-6 URL