Plant immune inducer ZNC promotes rutin accumulation and enhances resistance to Botrytis cinerea in tomato

Abstract

Gray mold is a destructive disease caused by Botrytis cinerea, a pervasive plant pathogen, which poses a threat to both tomato growth and postharvest storage. The utilization of induced resistance presents a potential strategy for combating plant pathogenic attacks. ZNC (zhinengcong), an extract derived from the endophytic fungus Paecilomyces variotii, has been discovered to play a vital role in preventing diverse forms of bacterial infections. Nevertheless, the precise mechanism behind its ability to enhance tomato resistance to fungi remains unclear. In this study, we found that the exogenous spraying of ZNC could significantly improve the resistance of tomato plants to B. cinerea. The results of both the metabolomic analysis and high-performance liquid chromatography (HPLC) demonstrated that tomato plants responded to ZNC treatment by accumulating high levels of rutin. Additional transcriptome analysis uncovered that rutin enhances tomato resistance possible by initiating the generation of reactive oxygen species (ROS) and phosphorylation of mitogen-activated protein kinases (MPKs) related genes expression during the initial phase of invasion by B. cinerea. In addition, we also found that rutin might activate plant immunity by eliciting ethylene (ET) and jasmonic acid (JA)-mediated pathways. Therefore, plant immune inducer ZNC and rutin has bright application prospects and high utilization value to control gray mold.

Keywords

Endophytic fungus extract / Metabolome / Flavonoids / Tomato gray mold / JA Signaling / ROS

References

[1] Adnan M, Hamada MS, Li GQ, Luo CX (2018) Detection and molecular characterization of resistance to the dicarboximide and benzamide fungicides in Botrytis cinerea from tomato in Hubei Province. China Plant Disease 102(7):1299–1306. https://doi.org/10.1094/PDIS-10-17-1531-RE

[2] BikaR, Baysal-GurelF, JenningsC. Botrytis cinerea management in ornamental production: a continuous battle. Can J Plant Path, 2021, 43(3):345-365

[3] CaarlsL, PieterseCM, Van WeesSC. How salicylic acid takes transcriptional control over jasmonic acid signaling. Front Plant Sci, 2015, 6: 170

[4] CaoJ, LiuB, XuX, ZhangX, ZhuC, LiY, et al.. Plant endophytic fungus extract ZNC improved potato immunity, yield, and quality. Front Plant Sci, 2021, 12: 707256

[5] ChuaLS. A review on plant-based rutin extraction methods and its pharmacological activities. J Ethnopharmacol, 2013, 150(3):805-817

[6] CourbierS, SnoekBL, KajalaK, LiL, van WeesSCM, PierikR. Mechanisms of far-red light-mediated dampening of defense against Botrytis cinerea in tomato leaves. Plant Physiol, 2021, 187(3):1250-1266

[7] De GeyterN, GholamiA, GoormachtigS, GoossensA. Transcriptional machineries in jasmonate-elicited plant secondary metabolism. Trends Plant Sci, 2012, 17(6):349-359

[8] DongX. SA, JA, ethylene, and disease resistance in plants. Curr Opin Plant Biol, 1998, 1(4):316-323

[9] DongNQ, SunY, GuoT, ShiCL, ZhangYM, KanY, et al.. UDP-glucosyltransferase regulates grain size and abiotic stress tolerance associated with metabolic flux redirection in rice. Nat Commun, 2020, 11(1):2629

[10] DouL, SunY, LiS, GeC, ShenQ, LiH, et al.. Transcriptomic analyses show that 24-epibrassinolide (EBR) promotes cold tolerance in cotton seedlings. PLoS ONE, 2021, 16(2):e0245070

[11] ErbM, KliebensteinDJ. Plant secondary metabolites as defenses, regulators, and primary metabolites: the blurred functional trichotomy. Plant Physiol, 2020, 184(1):39-52

[12] FinitiI, de laO, LeyvaM, VicedoB, Gómez‐PastorR, López‐CruzJ, García‐AgustínP, et al.. Hexanoic acid protects tomato plants against Botrytis cinerea by priming defence responses and reducing oxidative stress. Mol Plant Pathol, 2014, 15(6):550-562

[13] González-DomínguezE, FedeleG, CaffiT, DelièreL, SaurisP, GramajeD, et al.. A network meta-analysis provides new insight into fungicide scheduling for the control of Botrytis cinerea in vineyards. Pest Manag Sci, 2019, 75(2):324-332

[14] GovrinEM, LevineA. The hypersensitive response facilitates plant infection by the necrotrophic pathogen Botrytis cinerea. Curr Biol, 2000, 10(13):751-757

[15] HammerschmidtR. Induced disease resistance: how do induced plants stop pathogens?. Physiol Mol Plant Pathol, 1999, 55(2):77-84

[16] JantrawutP, PhongpradistR, MullerM, ViernsteinH. Enhancement of anti-inflammatory activity of polyphenolic flavonoid rutin by encapsulation. Pak J Pharm Sci, 2017, 30(5):1521-1528

[17] JonesJD, DanglJL. The plant immune system. Nature, 2006, 444(7117):323-329

[18] LiN, LinB, WangH, LiX, YangF, DingX, et al.. Natural variation in ZmFBL41 confers banded leaf and sheath blight resistance in maize. Nat Genet, 2019, 51(10):1540-1548

[19] Li R, Wang L, Li Y, Zhao R, Zhang Y, Sheng J et al (2020) Knockout of SlNPR1 enhances tomato plants resistance against Botrytis cinerea by modulating ROS homeostasis and JA/ET signaling pathways. Physiol Plant 170(4):569–579. https://doi.org/10.1111/ppl.13194

[20] LimW, LiJ. Synergetic effect of the Onion CHI gene on the PAP1 regulatory gene for enhancing the flavonoid profile of tomato skin. Sci Rep, 2017, 7(1):12377

[21] LiuXL, WangL, WangXW, YanY, YangXL, XieMY, et al.. Mutation of the chloroplast-localized phosphate transporter OsPHT2;1 reduces flavonoid accumulation and UV tolerance in rice. Plant J, 2020, 102(1):53-67

[22] LiuJ, ShenY, CaoH, HeK, ChuZ, LiN. OsbHLH057 targets the AATCA cis-element to regulate disease resistance and drought tolerance in rice. Plant Cell Rep, 2022, 12(5):1285-1299

[23] LuC, LiuH, JiangD, WangL, JiangY, TangS, et al.. Paecilomyces variotii extracts (ZNC) enhance plant immunity and promote plant growth. Plant Soil, 2019, 441(1):383-397

[24] MaZC, SongTQ, ZhuL, YeWW, WangY, ShaoYY, et al.. A Phytophthora sojae glycoside hydrolase 12 protein is a major virulence factor during soybean infection and is recognized as a PAMP. Plant Cell, 2015, 27(7):2057-2072

[25] MarschallR, TudzynskiP. Reactive oxygen species in development and infection processes. Semin Cell Dev Biol, 2016, 57: 138-146

[26] MehdyMC. Active oxygen species in plant defense against pathogens. Plant Physiol, 1994, 105(2):467

[27] MellidouI, KoukounarasA, KostasS, PatelouE, KanellisAK. Regulation of vitamin C accumulation for improved tomato fruit quality and alleviation of abiotic Stress. Genes (basel), 2021, 12(5):694

[28] MengF, YangC, CaoJ, ChenH, PangJ, ZhaoQ, et al.. A bHLH transcription activator regulates defense signaling by nucleo-cytosolic trafficking in rice. J Integr Plant Biol, 2020, 62(10):1552-1573

[29] MosbachA, EdelD, FarmerAD, WiddisonS, BarchiettoT, DietrichRA, et al.. Anilinopyrimidine resistance in Botrytis cinerea is linked to mitochondrial function. Front Microbiol, 2017, 8: 236

[30] NávarováH, BernsdorffF, DöringAC, ZeierJ. Pipecolic acid, an endogenous mediator of defense amplification and priming, is a critical regulator of inducible plant immunity. Plant Cell, 2012, 24(12):5123-5141

[31] NieJ, ZhouW, LiuJ, TanN, ZhouJM, HuangL. A receptor-like protein from Nicotiana benthamiana mediates VmE02 PAMP-triggered immunity. New Phytol, 2021, 229(4):2260-2272

[32] ParkerJE, HesslerG, CuiH. A new biochemistry connecting pathogen detection to induced defense in plants. New Phytol, 2022, 234(3):819-826

[33] PengC, ZhangA, WangQ, SongY, ZhangM, DingX, et al.. Ultrahigh-activity immune inducer from endophytic Fungi induces tobacco resistance to virus by SA pathway and RNA silencing. BMC Plant Biol, 2020, 20(1):169

[34] PengY, YangJ, LiX, ZhangY. Salicylic acid: biosynthesis and signaling. Annu Rev Plant Biol, 2021, 72: 761-791

[35] QuS, DaiC, GuoH, WangC, HaoZ, TangQ, et al.. Rutin attenuates vancomycin-induced renal tubular cell apoptosis via suppression of apoptosis, mitochondrial dysfunction, and oxidative stress. Phytother Res, 2019, 33(8):2056-2063

[36] Ramirez-PradoJS, AbulfarajAA, RayapuramN, BenhamedM, HirtH. Plant immunity: from signaling to epigenetic control of defense. Trends Plant Sci, 2018, 23(9):833-844

[37] RanfS, GischN, SchafferM, IlligT, WestphalL, KnirelYA, et al.. A lectin S-domain receptor kinase mediates lipopolysaccharide sensing in Arabidopsis thaliana. Nat Immunol, 2015, 16(4):426-433

[38] Rossi FR, Gárriz A, Marina M, Romero FM, Gonzalez ME, Collado IG et al (2011) The sesquiterpene botrydial produced by Botrytis cinerea induces the hypersensitive response on plant tissues and its action is modulated by salicylic acid and jasmonic acid signaling. Mol Plant Microbe Interact 24(8):888–896. https://doi.org/10.1094/mpmi-10-10-0248

[39] ShenX, WangR, XiongX, et al.. Metabolic reaction network-based recursive metabolite annotation for untargeted metabolomics. Nat Commun, 2019, 10(1):1516

[40] Shu P, Zhang S, Li Y, Wang X, Yao L, Sheng J et al (2021) Over-expression of SlWRKY46 in tomato plants increases susceptibility to Botrytis cinerea by modulating ROS homeostasis and SA and JA signaling pathways. Plant Physiol Biochem 166:1–9. https://doi.org/10.1016/j.plaphy.2021.05.021

[41] Soltis NE, Atwell S, Shi G, Fordyce R, Gwinner R, Gao D et al (2019) Interactions of tomato and Botrytis cinerea genetic diversity: parsing the contributions of host differentiation, domestication, and pathogen variation. Plant Cell 31(2):502–519. https://doi.org/10.1105/tpc.18.00857

[42] SongJ, BianJ, XueN, XuY, WuJ. Inter-species mRNA transfer among green peach aphids, dodder parasites, and cucumber host plants. Plant Diversity, 2022, 44(1):1-10

[43] SuP, ZhaoL, LiW, ZhaoJ, YanJ, MaX, et al.. Integrated metabolo-transcriptomics and functional characterization reveals that the wheat auxin receptor TIR1 negatively regulates defense against Fusarium graminearum. J Integr Plant Biol, 2021, 63(2):340-352

[44] TestoneG, MeleG, di GiacomoE, TenoreGC, GonnellaM, NicolodiC, et al.. Transcriptome driven characterization of curly- and smooth-leafed endives reveals molecular differences in the sesquiterpenoid pathway. Horticulture Res, 2019, 6: 1

[45] UngerC, KletaS, JandlG, TiedemannA, v. . Suppression of the defence-related oxidative burst in bean leaf tissue and bean suspension cells by the necrotrophic pathogen Botrytis cinerea. J Phytopathol, 2005, 153(1):15-26

[46] VatsS, BansalR, RanaN, KumawatS, BhattV, JadhavP, et al.. Unexplored nutritive potential of tomato to combat global malnutrition. Crit Rev Food Sci Nutr, 2022, 62(4):1003-1034

[47] Vuorinen K, Zamora O, Vaahtera L, Overmyer K, Brosché M (2021) Dissecting contrasts in cell death, hormone, and defense signaling in response to Botrytis cinerea and reactive oxygen species. Mol Plant Microbe Interact 34(1):75–87. https://doi.org/10.1094/MPMI-07-20-0202-R

[48] WangP, ZhouL, JamiesonP, ZhangL, ZhaoZ, BabiloniaK, et al.. The cotton wall-associated kinase GhWAK7A mediates responses to fungal wilt pathogens by complexing with the chitin sensory receptors. Plant Cell, 2020, 32(12):3978-4001

[49] WuT, ZhangH, YuanB, LiuH, KongL, ChuZ, et al.. Tal2b targets and activates the expression of OsF3H_03g to hijack OsUGT74H4 and synergistically interfere with rice immunity. New Phytol, 2022, 233(4):1864-1880

[50] YamaguchiT, YamadaA, HongN, OgawaT, IshiiT, ShibuyaN. Differences in the recognition of glucan elicitor signals between rice and soybean: beta-glucan fragments from the rice blast disease fungus Pyricularia oryzae that elicit phytoalexin biosynthesis in suspension-cultured rice cells. Plant Cell, 2000, 12(5):817-826

[51] YangW, XuX, LiY, WangY, LiM, WangY, et al.. Rutin-mediated priming of plant resistance to three bacterial pathogens initiating the early SA signal pathway. PLoS ONE, 2016, 11(1):e0146910

[52] YangY, WangX, ChenP, ZhouK, XueW, AbidK, et al.. Redox status, JA and ET signaling pathway regulating responses to Botrytis cinerea infection between the resistant cucumber genotype and its susceptible mutant. Front Plant Sci, 2020, 11: 559070

[53] YangL, KangY, LiuJ, LiN, SunH, AoT, et al.. Foliar spray with rutin improves cadmium remediation efficiency excellently by enhancing antioxidation and phytochelatin detoxification of Amaranthus hypochondriacus. Int J Phytorem, 2022, 24(10):1060-1070

[54] YuIC, ParkerJ, BentAF. Gene-for-gene disease resistance without the hypersensitive response in Arabidopsis dnd1 mutant. Proc Natl Acad Sci, 1998, 95(13):7819-7824

[55] YuanM, NgouBPM, DingP, XinXF. PTI-ETI crosstalk: an integrative view of plant immunity. Curr Opinion Plant Biol, 2021, 62: 102030

[56] ZhanC, LeiL, LiuZ, ZhouS, YangC, ZhuX, et al.. Selection of a subspecies-specific diterpene gene cluster implicated in rice disease resistance. Nat Plants, 2020, 6(12):1447-1454

[57] ZhanC, ShenS, YangC, LiuZ, FernieAR, GrahamIA, et al.. Plant metabolic gene clusters in the multi-omics era. Trends Plant Sci, 2022, 27(10):981-1001

[58] ZhangJ, CoakerG, ZhouJM, DongX. Plant immune mechanisms: from reductionistic to holistic points of view. Mol Plant, 2020, 13(10):1358-1378

[59] ZhengX, LiY, XiX, MaC, SunZ, YangX, et al.. Exogenous Strigolactones alleviate KCl stress by regulating photosynthesis, ROS migration and ion transport in Malus hupehensis Rehd. Plant Physiol Biochem, 2021, 159: 113-122

[60] ZhouJM, ZhangY. Plant immunity: danger perception and signaling. Cell, 2020, 181(5):978-989

Funding

the National Key Research and Development Program(2022YFD1401500); major Basic Research Project of Natural Science Foundation of Shandong Province(ZR2022ZD23); the National Natural Science Foundation(32272557); Shandong Modern Agricultural Industry Technology System(SDAIT-04-08); Shandong Province Key Research and Development Plan(2021TZXD007-04-4)