Integrated evaluation of antifungal activity of pomegranate peel polyphenols against a diverse range of postharvest fruit pathogens

Abstract

The search for safe, natural antimicrobial agents has received significant attention for controlling postharvest diseases in fruits. This study evaluated the antifungal activity of pomegranate peel extracts (PPE) against 9 pathogenic fungi by different methods. Additionally, the key antifungal polyphenol compounds were identified. Results revealed that the n-hexane partitioned fraction of ethanolic extract exhibited the highest inhibition efficacy. A total 36 polyphenols, including 10 newly discovered compounds in pomegranate peel were recognized. Among these, 9 specific polyphenols were found to be relatively abundant in the n-hexane fraction, highlighting their potential as primary antifungal agents. Notably, the newly identified polyphenol compound nobiletin demonstrated the strongest inhibitory effects against Colletotrichum gloeosporioides, Rhizopus stolonifer and Aspergillus niger with inhibition diameters of 12.2, 12.0, and 12.5 mm, respectively. Salidroside exhibited significant antifungal activity against Monilinia fructicola and A. niger (inhibition diameters > 12.0 mm). Furthermore, cinnamic acid that was enriched in hexane fraction, showed the highest inhibitory zones against Alternaria alternata, C. gloeosporioides and Botryosphaeria dothidea (11.8 ~ 12.7 mm). Caffeic acid and protocatechuic acid both displayed the lowest minimum inhibitory concentration (MIC) on Penicillium expansum. The study demonstrated the potential of PPE, particularly the n-hexane fraction, as a broad-spectrum natural antifungal agent for postharvest disease management.

Keywords

Pomegranate peel / Polyphenols / Antifungal activity / Fruit / Postharvest pathogens

References

[1] AbdullaR, MansurS, LaiH, UbulA, SunG, HuangG, AisaHA. Qualitative analysis of polyphenols in macroporous resin pretreated pomegranate husk extract by HPLC-QTOF‐MS. Phytochem Anal, 2017, 285465-473.

[2] AkhtarS, IsmailT, FraternaleD, SestiliP. Pomegranate Peel and Peel extracts: chemistry and food features. Food Chem, 2015, 174: 417-425.

[3] AldarhamiA, BazaidAS, AlhamedAS, AlghaithAF, AhamadSR, AlassmrryYA, AlharaziT, SnoussiM, QanashH, AlamriA, BadraouiR. Antimicrobial potential of Pithecellobium Dulce seed extract against pathogenic bacteria: in Silico and in vitro evaluation. Biomed Res Int, 2023, 202312848198.

[4] AmbigaipalanP, de CamargoAC, ShahidiF. Phenolic compounds of pomegranate byproducts (outer skin, mesocarp, divider membrane) and their antioxidant activities. J Agric Food Chem, 2016, 64346584-6604.

[5] Bautista-Baños S, Bosquez-Molina E, Barrera-Necha LL (2014) Rhizopus stolonifer (soft rot). In: Postharvest decay, PP 1–44. Elsevier. https://doi.org/10.1016/B978-0-12-411552-1.00001-6

[6] BrighentiV, IseppiR, PinziL, MincuzziA, IppolitoA, MessiP, SanzaniSM, RastelliG, PellatiF. Antifungal activity and DNA topoisomerase Inhibition of hydrolysable tannins from Punica granatum L. Int J Mol Sci, 2021, 2284175.

[7] DengY, LiY, YangF, ZengA, YangS, LuoY, ZhangY, XieY, YeT, XiaY. The extract from Punica granatum (pomegranate) Peel induces apoptosis and impairs metastasis in prostate cancer cells. Biomed Pharmacother, 2017, 93: 976-984.

[8] DeyD, DebnathS, HazraS, GhoshS, RayR, HazraB. Pomegranate pericarp extract enhances the antibacterial activity of Ciprofloxacin against extended-spectrum β-lactamase (ESBL) and metallo-β-lactamase (MBL) producing Gram-negative bacilli. Food Chem Toxicol, 2012, 50124302-4309.

[9] DikbasN, KotanR, DadasogluF, SahinF. Control of Aspergillus flavus with essential oil and methanol extract of Satureja hortensis. Int J Food Microbiol, 2008, 1242179-182.

[10] El-HadaryAE, RamadanMF. Phenolic profiles, antihyperglycemic, antihyperlipidemic, and antioxidant properties of pomegranate (Punica granatum) Peel extract. J Food Biochem, 2019, 434e12803.

[11] Elkahoui S, Jamal A, Ben Abbes W, Gzara L, Charradi K, Haddaji N, Kachout M, Abdelgadir A, Bardakci F, Badraoui R (2024) Investigation of the valorization of methanolic extract of Punica granatum L. peel in terms of phytochemical, trace element, antioxidant activities and ADMET profile of active compounds. Pol J Environ Stud 33(3). https://doi.org/10.15244/pjoes/175022

[12] EloffJN. Avoiding pitfalls in determining antimicrobial activity of plant extracts and publishing the results. BMC Complement Altern Med, 2019, 19: 1-8.

[13] Fang J, Luo L, Li W, Li H (2025) Application of novel adsorbents in methanol removal from ethanol: Citric acid-modified 4A zeolites and Cu@ZIF-7. Biochem Eng J. https://doi.org/10.1016/j.bej.2025.109719

[14] FossSR, NakamuraCV, Ueda-NakamuraT, CortezDA, EndoEH, Dias FilhoBP. Antifungal activity of pomegranate Peel extract and isolated compound Punicalagin against dermatophytes. Ann Clin Microbiol Antimicrob, 2014, 13: 1-6.

[15] GarganeseF, SanzaniSM, Di RellaD, SchenaL, IppolitoA. Pre-and postharvest application of alternative means to control Alternaria brown spot of citrus. Crop Prot, 2019, 121: 73-79.

[16] GlazerI, MasaphyS, MarcianoP, Bar-IlanI, HollandD, KeremZ, AmirR. Partial identification of antifungal compounds from Punica granatum Peel extracts. J Agric Food Chem, 2012, 160194841-4848.

[17] GomesA, QueirozM, PereiraO. Mycofumigation for the biological control of post-harvest diseases in fruits and vegetables: a review. Austin J Biotechnol Bioeng, 2015, 241-8

[18] GonelimaliFD, LinJ, MiaoW, XuanJ, CharlesF, ChenM, HatabSR. Antimicrobial properties and mechanism of action of some plant extracts against food pathogens and spoilage microorganisms. Front Microbiol, 2018, 9: 1639.

[19] Gosset-ErardC, ZhaoM, Lordel-MadeleineS, EnnaharS. Identification of Punicalagin as the bioactive compound behind the antimicrobial activity of pomegranate (Punica granatum L.) peels. Food Chem, 2021, 352: 129396.

[20] HeX, WangL, XiaB, CaoX, HuN, HuangJ, YiY. Antifungal effect of cinnamic acid and induced resistance of cinnamic acid-protocatechuic acid-CaCl₂-NaCl-pullulan composite preservative to Trichoderma Harzianum in postharvest Hypsizygus marmoreus. LWT-Food Sci Technol, 2023, 15184115108.

[21] HemegHA, MoussaIM, IbrahimS, DawoudTM, AlhajiJH, MubarakAS, KabliSA, AlsubkiRA, TawfikAM, MaroufSA. Antimicrobial effect of different herbal plant extracts against different microbial population. Saudi J Biol Sci, 2020, 27123221-3227.

[22] HernándezA, Ruiz-MoyanoS, GalvánAI, MerchánAV, NevadoFP, ArandaE, SerradillaMJ, de Guía CórdobaM, MartínA. Anti-fungal activity of phenolic sweet orange Peel extract for controlling fungi responsible for post-harvest fruit decay. Fungal Biol, 2021, 1252143-152.

[23] IbrahiumM. Efficiency of pomegranate Peel extract as antimicrobial, antioxidant and protective agents. World J Agric Sci, 2010, 64338-344

[24] JebahiS, Ben SalahG, JarrayS, NaffatiM, AhmadMA, BrahmiF, SaeedM, SiddiquiAJ, AbdelmajidK, BadraouiR. Chitosan-based gastric dressing materials loaded with pomegranate Peel as bioactive agents: pharmacokinetics and effects on experimentally induced gastric ulcers in rabbits. Metabolites, 2022, 12121158.

[25] Kahramanoğlu İ (2024) Postharvest challenges, chemical control of postharvest diseases and concerns over agrochemicals. In: Postharvest Physiology and Handling of Horticultural Crops CRC, pp 85–101

[26] KulkarniAP, MahalH, KapoorS, AradhyaS. In vitro studies on the binding, antioxidant, and cytotoxic actions of Punicalagin. J Agric Food Chem, 2007, 5541491-1500.

[27] KumarV, SoniR, NathawatB. Eco-friendly management of Aspergillus rot of pomegranate (Punica granatum). Adv Life Sci, 2016, 572859-2865

[28] LastochkinaO, BaymievA, ShayahmetovaA, GarshinaD, KoryakovI, ShpirnayaI, PusenkovaL, Mardanshin Id, KasnakC, PalamutogluR. Effects of endophytic Bacillus subtilis and Salicylic acid on postharvest diseases (Phytophthora infestans, Fusarium oxysporum) development in stored potato tubers. Plants, 2020, 9176.

[29] LattanzioV, CardinaliA, PalmieriS. The role of phenolics in the postharvest physiology of fruits and vegetables: Browning reaction and fungal diseases. Ital J Food Sci, 1994, 6: 3-22

[30] LiuH, LiuJ, LvZ, YangW, ZhangC, ChenD, JiaoZ. Effect of dehydration techniques on bioactive compounds in Hawthorn slices and their correlations with antioxidant properties. J Food Sci Technol, 2019, 56: 2446-2457.

[31] LiuH, LvZ, YangW, LiA, LiuJ, ZhangQ, JiaoZ. Virtual cold chain method to evaluate the effect of rising temperature on the quality evolution of Peach fruit. Foods, 2023, 12122403.

[32] LouZ, WangH, RaoS, SunJ, MaC, LiJ. -Coumaric acid kills bacteria through dual damage mechanisms. Food Control, 2012, 252550-554.

[33] Lv Z, Liu H, Yang W, Zhang Q, Chen D, Jiao Z, Liu J (2024) Comprehensive analysis of physicochemical properties and volatile compounds in different strawberry wines under various pre-treatments. Molecules 29(9): 2045. https://doi.org/10.3390/molecules29092045

[34] ManG, MaY, XuL, LiaoX, ZhaoL. Comparison of thermal and non-thermal extraction methods on free and bound phenolics in pomegranate Peel. Innov Food Sci Emerg Technol, 2023, 84: 103291.

[35] MoralesJ, MendozaL, CotorasM. Alteration of oxidative phosphorylation as a possible mechanism of the antifungal action of p-coumaric acid against Botrytis cinerea. J Appl Microbiol, 2017, 1234969-976.

[36] NicosiaMGLD, PangalloS, RaphaelG, RomeoFV, StranoMC, RapisardaP, DrobyS, SchenaL. Control of postharvest fungal rots on citrus fruit and sweet cherries using a pomegranate Peel extract. Postharvest Biol Technol, 2016, 114: 54-61.

[37] NoumiE, AhmadI, BoualiN, PatelH, GhannayS, ALrashidiAA, AbdulhakeemMA, PatelM, CeylanO, BadraouiR, Mousa ElayyanAE. Thymus musilii Velen. Methanolic extract: in vitro and in Silico screening of its antimicrobial, antioxidant, anti-quorum sensing, antibiofilm, and anticancer activities. Life, 2022, 13162.

[38] NunesCA. Biological control of postharvest diseases of fruit. Eur J Plant Pathol, 2012, 133: 181-196.

[39] ObiVI, BarriusoJJ, GogorcenaY. Effects of pH and titratable acidity on the growth and development of Monilinia Laxa (Aderh. & Ruhl.) in vitro and in vivo. Eur J Plant Pathol, 2018, 151: 781-790.

[40] OrgilO, SchwartzE, BaruchL, MatityahuI, MahajnaJ, AmirR. The antioxidative and anti-proliferative potential of non-edible organs of the pomegranate fruit and tree. LWT-Food Sci Technol, 2014, 582571-577.

[41] PaneC, FratianniF, ParisiM, NazzaroF, ZaccardelliM. Control of alternaria post-harvest infections on Cherry tomato fruits by wild pepper phenolic-rich extracts. Crop Prot, 2016, 84: 81-87.

[42] ParkG-G, KimW, YangK-Y. Rapid and sensitive detection of the causal agents of postharvest Kiwifruit rot, Botryosphaeria Dothidea and Diaporthe eres, using a recombinase polymerase amplification assay. Plant Pathol J, 2023, 395522.

[43] PerumalS, MahmudR, IsmailS. Mechanism of action of isolated caffeic acid and epicatechin 3-gallate from Euphorbia Hirta against Pseudomonas aeruginosa. Pharmacogn Mag, 2017, 13Suppl 2S311-S315.

[44] QiuP, MiA, HongC, HuangS, MaQ, LuoQ, QiuJ, JiangH, ChenY, ChenF, YanH, ZhaoJ, KongY, DuY, LiC, KongD, EfferthT, LouD. An integrated network Pharmacology approach reveals that Ampelopsis grossedentata improves alcoholic liver disease via TLR4/NF-κB/MLKL pathway. Phytomedicine, 2024, 132: 155658.

[45] Rosas-Burgos EC, Cortez-Rocha MO, Cinco-Moroyoqui FJ, Robles-Zepeda RE, López-Cervantes J, Sánchez-Machado DI, Lares-Villa F (2009) Antifungal activity in vitro of Baccharis glutinosa and Ambrosia confertiflora extracts on Aspergillus flavus, Aspergillus parasiticus and Fusarium verticillioides. World J Microbiol Biotechnol 25: 2257–2261. https://doi.org/10.1007/s11274-009-0116-1

[46] Rosas-BurgosEC, Burgos‐HernándezA, Noguera‐ArtiagaL, KačániováM, Hernández‐GarcíaF, Cárdenas‐LópezJL, Carbonell‐BarrachinaÁA. Antimicrobial activity of pomegranate Peel extracts as affected by cultivar. J Sci Food Agric, 2017, 973802-810.

[47] SalimA, DeianaP, FancelloF, MolinuMG, SantonaM, ZaraS. Antimicrobial and antibiofilm activities of pomegranate Peel phenolic compounds: varietal screening through a multivariate approach. J Bioresour Bioprod, 2023, 82146-161.

[48] SorrentiV, RandazzoCL, CaggiaC, BallistreriG, RomeoFV, FabroniS, TimpanaroN, RaffaeleM, VanellaL. Beneficial effects of pomegranate Peel extract and probiotics on pre-adipocyte differentiation. Front Microbiol, 2019, 10: 660.

[49] SreekumarS, SithulH, MuraleedharanP, AzeezJM, SreeharshanS. Pomegranate fruit as a rich source of biologically active compounds. Biomed Res Int, 2014, 20141686921.

[50] SweidanN, Abu RayyanW, MahmoudI, AliL. Phytochemical analysis, antioxidant, and antimicrobial activities of Jordanian pomegranate peels. PLoS ONE, 2023, 1811e0295129.

[51] TehranifarA, SelahvarziY, KharraziM, BakhshVJ. High potential of agro-industrial by-products of pomegranate (Punica granatum L.) as the powerful antifungal and antioxidant substances. Ind Crops Prod, 2011, 3431523-1527.

[52] TengH, DengH, WuY, ZhangC, AiC, CaoH, XiaoJ, ChenL. Effects of Rhodiola rosea and its major compounds on insulin resistance in Caenorhabitis elegans. J Future Foods, 2022, 24365-371.

[53] VelazhahanR. Potential use of Zataria multiflora essential oil to control postharvest Aspergillus flavus fruit rot of strawberry. J Agric Mar Sci, 2023, 28238-44.

[54] Viuda-MartosM, Fernández‐LópezJ, Pérez‐ÁlvarezJ. Pomegranate and its many functional components as related to human health: a review. Compr Rev Food Sci Food Saf, 2010, 96635-654.

[55] WangM, JiangN, WangY, JiangD, FengX. Characterization of phenolic compounds from early and late ripening sweet cherries and their antioxidant and antifungal activities. J Agric Food Chem, 2017, 65265413-5420.

[56] WangL, HouH, ZhouZ, TuH, YuanH. Identification and detection of Botryosphaeria Dothidea from Kiwifruit (Actinidia chinensis) in China. Plants, 2021, 102401.

[57] YangW, LiuJ, LiuH, ZhangQ, LvZ, JiaoZ. Characterization of strawberry purees fermented by Lactobacillus spp. Based on nutrition and flavor profiles using LC-TOF/MS, HS-SPME-GC/MS and E-nose. LWT-Food Sci Technol, 2023, 189: 15457.

[58] YangW, LvZ, LiuH, ZhangQ, QiaoC, NawazM, JiaoZ, LiuJ. Effect of organic acid addition before fermentation on the physicochemical and sensory properties of Cherry wine. Foods, 2024, 13233902.

[59] Zaushintsena A, Babich O, Bryukhachev E, Milenteva I, Starcev A (2020) Opportunities for using biologically active substances Rhodiola Rosea L. in the production of functional food with consideration for antimicrobial activity. In: E3S Web of Conferences, 2020. EDP Sciences, p 01011. https://doi.org/10.1051/e3sconf/202017601011

[60] ZhangH, GodanaEA, SuiY, YangQ, ZhangX, ZhaoL. Biological control as an alternative to synthetic fungicides for the management of grey and blue mould diseases of table grapes: a review. Crit Rev Microbiol, 2020, 464450-462.

[61] ZhangQ, YangW, LiuJ, LiuH, LvZ, ZhangC, ChenD, JiaoZ. Postharvest UV-C irradiation increased the flavonoids and anthocyanins accumulation, phenylpropanoid pathway gene expression, and antioxidant activity in sweet cherries (Prunus avium L). Postharvest Biol Technol, 2021, 175: 111490.

[62] ZhaoS, GuoY, WangQ, LuoH, HeC, AnB. Expression of Flagellin at yeast surface increases biocontrol efficiency of yeast cells against postharvest disease of tomato caused by Botrytis cinerea. Postharvest Biol Technol, 2020, 162: 111112.

[63] ZhengL, SituJ-j, ZhuQ-f, XiP-g, ZhengY, LiuH-x, ZhouX. Identification of volatile organic compounds for the biocontrol of postharvest Litchi fruit pathogen Peronophythora Litchii. Postharvest Biol Technol, 2019, 155: 37-46.

Funding

National Key Research and Development Program(2022YFD1600700)

Agricultural Science and Technology Innovation Program of the Chinese Academy of Agricultural Sciences (CAAS-ASTIP-2023-ZFRI)

RIGHTS & PERMISSIONS

The Author(s)