Abstract
Key message
This is the first report of the anti-oomycete activity of A. unedo leaves extract against Phytophthora cinnamomi. The extract was characterized by HPLC–MS and different chemotypes identified.
Abstract
Arbutus unedo L. is a small perennial Ericaceae tree well adapted to the environmental conditions of its Mediterranean distribution area. It produces a wide range of secondary metabolites with bioactive properties and applications for food, cosmetic and pharmaceutical industries. Due to the antifungal activity of some secondary metabolites they can play a key role in plant defense. In particular, arbutin and hydroquinone, two phenolic compounds typical of the Ericaceae species, might be especially important in plant defense against pathogens. Thus, in this work we (i) established a phenolic fingerprint of wild strawberry tree leaves, (ii) evaluated the effect of genotype and seasonal variation on phenolic metabolite composition, (iii) quantified arbutin and hydroquinone, and (iv) tested the antifungal effect of arbutin, hydroquinone and a methanol extract of A. unedo leaves against common strawberry tree pathogens. A total of 54 compounds were assigned by HPLC–PDA–ESI/MSn, namely, gallic acid derivatives, hydrolysable and condensed tannins, flavonoids, arbutin, hydroquinone and their derivatives, and significant differences on relative abundance of the phenolic compounds between individuals were verified. Hydroquinone and its glucoside arbutin were quantified and its antifungal and anti-oomycete effect against A. unedo pathogens evaluated. Arbutin concentrations ranged from 0.69 ± 0.05 to 22.97 ± 0.76 mg g−1 fresh weight of leaves. Hydroquinone was only detected in a few samples and its concentration ranged from 0.069 ± 0.004 to 0.604 ± 0.024 mg g−1 fresh weight. Both arbutin and hydroquinone induced a reduction in growth of Glomerella cingulata and both compounds were highly effective against Phytophthora cinnamomi, with a reduction in growth close to 80% at the higher arbutin concentration tested. Microplate tests showed that a methanol extract of A. unedo leaves displayed an anti-oomycete activity with MIC (minimum inhibitory concentration) against P. cinnamomi at an arbutin equivalent concentration of 0.625 mg mL−1, and was more active against P. cinnamomi than the arbutin standard (MIC = 12 mg mL−1), which suggests that a synergic effect occurs in the presence of other metabolites existing in the extract. Moreover, extract containing an arbutin equivalent concentration of 2.5 mg mL−1 was lethal for this pathogen. Arbutin, their derivatives and/or other secondary metabolites play an important role in A. unedo defense mechanisms against biotic stress, opening the way to biotechnological applications, namely, to use A. unedo leaf extracts as a natural phytosanitary agent. A genotype with the ability to produce high amounts of arbutin was also identified. The implications of these results on agroforestry management are discussed, as well as its relevance on plant phenotyping and selection for future breeding assays.
Similar content being viewed by others
References
Abdulla R, Mansur S, Lai H, Ubul A, Sun G, Huang G, Aisa HA (2017) Qualitative analysis of polyphenols in macroporous resin pretreated pomegranate husk extract by HPLC-QTOF-MS. Phytochem Anal 28:465–473. https://doi.org/10.1002/pca.2695
Abid M, Yaich H, Cheikhrouhou S, Khemakhem I, Bouaziz M, Attia H, Ayadi MA (2017) Antioxidant properties and phenolic profile characterization by LC–MS/MS of selected Tunisian pomegranate peels. J Food Sci Technol 54:2890–2901. https://doi.org/10.1007/s13197-017-2727-0
Alberti Á, Béni S, Lackó E, Riba P, Al-Khrasani M, Kéry Á (2012) Characterization of phenolic compounds and antinociceptive activity of Sempervivum tectorum L. leaf juice. J Pharm Biomed Anal 70:143–150. https://doi.org/10.1016/j.jpba.2012.06.017
Ali K, Maltese F, Figueiredo A, Rex M, Fortes AM, Zyprian E, Pais MS, Verpoorte R, Choi YH (2012) Alterations in grapevine leaf metabolism upon inoculation with Plasmopara viticola in different time-points. Plant Sci 191–192:100–107. https://doi.org/10.1016/j.plantsci.2012.04.014
Attard E (2013) A rapid microtitre plate Folin-Ciocalteu method for the assessment of polyphenols. Cent Eur J Biol 8:48–53. https://doi.org/10.2478/s11535-012-0107-3
Bollina V, Kumaraswamy GK, Kushalappa AC, Choo TM, Dion Y, Rioux S, Faubert D, Hamzehzarghani H (2010) Mass spectrometry-based metabolomics application to identify quantitative resistance-related metabolites in barley against Fusarium head blight. Mol Plant Pathol 11:769–782. https://doi.org/10.1111/j.1364-3703.2010.00643.x
Boulanouar B, Abdelaziz G, Aazza S, Gago C, Miguel MG (2013) Antioxidant activities of eight Algerian plant extracts and two essential oils. Ind Crops Prod 46:85–96. https://doi.org/10.1016/j.indcrop.2013.01.020
Clifford MN, Stoupi S, Kuhnert N (2007) Profiling and characterization by LC-MSn of the galloylquinic acids of green tea, tara tannin, and tannic acid. J Agric Food Chem 55:2797–2807. https://doi.org/10.1021/jf063533l
Coimbra AT, Luís ÂFS, Batista MT, Ferreira SMP, Duarte APC (2020) Phytochemical characterization, bioactivities evaluation and synergistic effect of Arbutus unedo and Crataegus monogyna extracts with Amphotericin B. Curr Microbiol 77:2143–2154. https://doi.org/10.1007/s00284-020-02125-w
Correia H, González-Paramás A, Amaral MT, Santos-Buelga C, Batista MT (2006) Polyphenolic profile characterization of Agrimonia eupatoria L. by HPLC with different detection devices. Biomed Chromatogr 20:88–94. https://doi.org/10.1002/bmc.533
Devillers J, Steiman R, Seigle-Muranti F (1989) The usefulness of the agar-well diffusion method for assessing chemical toxicity to bacteria and fungi. Chemosphere 19:1693–1700. https://doi.org/10.1016/0045-6535(89)90512-2
Fiorentino A, Castaldi S, D’Abrosca B, Natale A, Carfora A, Messere A, Monaco P (2007) Polyphenols from the hydroalcoholic extract of Arbutus unedo living in a monospecific Mediterranean woodland. Biochem Syst Ecol 35:809–811. https://doi.org/10.1016/j.bse.2007.04.005
Galeotti F, Barile E, Curir P, Dolci M, Lanzotti V (2008) Flavonoids from carnation (Dianthus caryophyllus) and their antifungal activity. Phytochem Lett 1:44–48. https://doi.org/10.1016/j.phytol.2007.10.001
Gan R-Y, Kong K-W, Li H-B, Wu K, Ge Y-Y, Chan C-L, Shi X-M, Corke H (2018) Separation, identification, and bioactivities of the main gallotannins of red sword bean (Canavalia gladiata) coats. Front Chem 6:1–10. https://doi.org/10.3389/fchem.2018.00039
Ganthaler A, Stöggl W, Kranner I, Mayr S (2017) Foliar phenolic compounds in Norway Spruce with varying susceptibility to Chrysomyxa rhododendri: analyses of seasonal and infection-induced accumulation patterns. Front Plant Sci 8:1173. https://doi.org/10.3389/fpls.2017.01173
Girardi FA, Tonial F, Chini SO, Sobottka AM, Scheffer-Basso SM, Bertol CD (2014) Phytochemical profile and antimicrobial properties of Lotus spp. (Fabaceae). An Acad Bras Cienc 86:1295–1302. https://doi.org/10.1590/0001-3765201420130220
Gu Z, R E, Schlesner M, (2016) Complex heatmaps reveal patterns and correlations in multidimensional genomic data. Bioinformatics. https://doi.org/10.1093/bioinformatics/btw313
Gunnaiah R, Kushalappa AC, Duggavathi R, Fox S, Somers DJ (2012) Integrated metabolo-proteomic approach to decipher the mechanisms by which wheat qtl (Fhb1) contributes to resistance against Fusarium graminearum. PLoS ONE 7:e40695. https://doi.org/10.1371/journal.pone.0040695
Hofmann T, Nebehaj E, Albert L (2016) Antioxidant properties and detailed polyphenol profiling of European hornbeam (Carpinus betulus L.) leaves by multiple antioxidant capacity assays and high-performance liquid chromatography/multistage electrospray mass spectrometry. Ind Crops Prod 87:340–349. https://doi.org/10.1016/j.indcrop.2016.04.037
Isah T (2019) Stress and defense responses in plant secondary metabolites production. Biol Res 52:39
Jang GH, Kim HW, Lee MK, Jeong SY, Bak AR, Lee DJ, Kim JB (2018) Characterization and quantification of flavonoid glycosides in the Prunus genus by UPLC-DAD-QTOF/MS. Saudi J Biol Sci 25:1622–1631. https://doi.org/10.1016/j.sjbs.2016.08.001
Jurica K, Karačonji IB, Šegan S, Opsenica DM, Kremer D (2015) Quantitative analysis of arbutin and hydroquinone in strawberry tree (Arbutus unedo L., Ericaceae) leaves by gas chromatography-mass spectrometry. Arh Hig Rada Toksikol 66:197–202. https://doi.org/10.1515/aiht-2015-66-2696
Jurica K, Brčić Karačonji I, Mikolić A, Milojković-Opsenica D, Benković V, Kopjar N (2018) In vitro safety assessment of the strawberry tree (Arbutus unedo L.) water leaf extract and arbutin in human peripheral blood lymphocytes. Cytotechnology 70:1261–1278. https://doi.org/10.1007/s10616-018-0218-4
Kang DS, Min KJ, Kwak AM, Lee SY, Kang HW (2017) Defense response and suppression of Phytophthora blight disease of pepper by water extract from spent mushroom substrate of Lentinula edodes. Plant Pathol J 33:264–275. https://doi.org/10.5423/PPJ.OA.02.2017.0030
Kuźniak E, Wielanek M, Chwatko G, Głowacki R, Libik-Konieczny M, Piatek M, Gajewska E, Skłodowska M (2015) Salicylic acid and cysteine contribute to arbutin-induced alleviation of angular leaf spot disease development in cucumber. J Plant Physiol 181:9–13. https://doi.org/10.1016/j.jplph.2015.03.017
Mariotto S, Esposito E, Di R, Ciampa A, Mazzon E, Carcereri A, Prati D, Darra E, Vincenzi S, Cucinotta G, Caminiti R, Suzuki H, Cuzzocrea S (2008) Protective effect of Arbutus unedo aqueous extract in carrageenan-induced lung inflammation in mice. Pharmacol Res 57:110–124. https://doi.org/10.1016/j.phrs.2007.12.005
Martins JF, Correia SI, Canhoto JM (2016) Somatic embryogenesis induction and plant regeneration in strawberry tree (Arbutus unedo L.). In: Germana M, Lambardi M (eds) Methods in molecular biology, vol 1359. Humana Press, New York, pp 329–339
Martins JF, Correia S, Correia B, Pinto G, Canhoto JM (2019) Shoot proliferation and organogenesis on Arbutus unedo: Physiological analysis under water stress. Biol Plant. 63:278–286. https://doi.org/10.32615/bp.2019.032
Mierziak J, Wojtasik W, Kostyn K, Czuj T, Szopa J, Kulma A (2014) Crossbreeding of transgenic flax plants overproducing flavonoids and glucosyltransferase results in progeny with improved antifungal and antioxidative properties. Mol Breed 34:1917–1932. https://doi.org/10.1007/s11032-014-0149-5
Miguel MG, Faleiro ML, Guerreiro AC, Antunes MD (2014) Arbutus unedo L.: chemical and biological properties. Molecules 19:15799–15823. https://doi.org/10.3390/molecules191015799
Moralejo E, Belbahri L, Calmin G, García-Muñoz JA, Lefort F, Descals E (2008) Strawberry tree blight in Spain, a new disease caused by various Phytophthora species. J Phytopathol 156:577–587. https://doi.org/10.1111/j.1439-0434.2008.01397.x
Moreira AC, Martins JMS (2005) Influence of site factors on the impact of Phytophthora cinnamomi in cork oak stands in Portugal. For Pathol 35:145–162. https://doi.org/10.1111/j.1439-0329.2005.00397.x
Morgado S, Morgado M, Plácido AI, Roque F, Duarte AP (2018) Arbutus unedo L.: from traditional medicine to potential uses in modern pharmacotherapy. J Ethnopharmacol 225:90–102. https://doi.org/10.1016/j.jep.2018.07.004
Munné-Bosch S, Peñuelas J (2004) Drought-induced oxidative stress in strawberry tree (Arbutus unedo L.) growing in Mediterranean field conditions. Plant Sci 166:1105–1110. https://doi.org/10.1016/j.plantsci.2003.12.034
Negri G, Tabach R (2013) Saponins, tannins and flavonols found in hydroethanolic extract from Periandra dulcis roots. Brazilian J Pharmacogn 23:851–860. https://doi.org/10.1590/S0102-695X2013000600001
Oliver AE, Hincha DK, Tsvetkova NM, Vigh L, Crowe JH (2001) The effect of arbutin on membrane integrity during drying is mediated by stabilization of the lamellar phase in the presence of nonbilayer-forming lipids. Chem Phys Lipids 111:37–57. https://doi.org/10.1016/S0009-3084(01)00141-4
Orak HH, Karamać M, Amarowicz R, Orak A, Penkacik K (2019) Genotype-related differences in the phenolic compound profile and antioxidant activity of extracts from olive (Olea europaea L.) leaves. Molecules 24:1130. https://doi.org/10.3390/molecules24061130
Padmavati M, Sakthivel N, Thara KV, Reddy AR (1997) Differential sensitivity of rice pathogens to growth inhibition by flavonoids. Phytochemistry 46:499–502. https://doi.org/10.1016/S0031-9422(97)00325-7
Parvez MM, Tomita-Yokotani K, Fujii Y, Konishi T, Iwashina T (2004) Effects of quercetin and its seven derivatives on the growth of Arabidopsis thaliana and Neurospora crassa. Biochem Syst Ecol 32:631–635. https://doi.org/10.1016/j.bse.2003.12.002
Pavarini DP, Pavarini SP, Niehues M, Lopes NP (2012) Exogenous influences on plant secondary metabolite levels. Anim Feed Sci Technol 176:5–16. https://doi.org/10.1016/j.anifeedsci.2012.07.002
Pavlović RD, Lakušić B, Došlov-Kokoruš Z, Kovačević N (2009) Arbutin content and antioxidant activity of some Ericaceae species. Pharmazie 64:656–659. https://doi.org/10.1691/ph.2009.9551
Polizzi G, Aiello D, Guarnaccia V, Vitale A, Perrone G, Stea G (2011) First report of damping-off on strawberry tree caused by Colletotrichum acutatum and C. simmondsii in Italy. Plant Dis 95:1588. https://doi.org/10.1094/PDIS-07-11-0567
Powell CC, Hildebrand DC (1970) Fire blight resistance in Pyrus: involvement of arbutin oxidation. Phytopathology 60:337–340. https://doi.org/10.1094/Phyto-60-337
R Core Team (2017) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. https://www.R-project.org/
Ravensdale M, Rocheleau H, Wang L, Nasmith C, Ouellet T, Subramaniam R (2014) Components of priming-induced resistance to Fusarium head blight in wheat revealed by two distinct mutants of Fusarium graminearum. Mol Plant Pathol 15:948–956. https://doi.org/10.1111/mpp.12145
Romero-Martin MA, Trapero-Casas A (2003) La mancha foliar del madroño (Arbutus unedo) causada por Septoria unedonis var. vellanensis. Bol Sanid Veg Plagas 29:375–392
Royse DJ, Ries SM (1978) The influence of fungi isolated from peach twigs on the pathogenicity of Cytospora cincta. Phytopathology 68:603–607. https://doi.org/10.1094/phyto-68-603
Rueden CT, Schindelin J, Hiner MC, DeZonia BE, Walter AE, Arena ET, Eliceiri KW (2017) Image J2: ImageJ for the next generation of scientific image data. BMC Bioinformatics 18:529. https://doi.org/10.1186/s12859-017-1934-z
Saldanha LL, Vilegas W, Dokkedal AL (2013) Characterization of flavonoids and phenolic acids in Myrcia bella Cambess. using FIA-ESI-IT-MSn and HPLC-PAD-ESI-IT-MS combined with NMR. Molecules 18:8402–8416. https://doi.org/10.3390/molecules18078402
Schloerke B, Crowley J, Cook D, Briatte F, Marbach M, Thoen E, Elberg A, Larmarange J (2018) GGally: Extension to “ggplot2”. R package version 1.4.0. https://CRAN.R-project.org/package=GGally
Sena K, Crocker E, Vincelli P, Barton C (2018) Phytophthora cinnamomi as a driver of forest change: Implications for conservation and management. For Ecol Manage 409:799–807
Sherwood P, Bonello P (2013) Austrian pine phenolics are likely contributors to systemic induced resistance against Diplodia pinea. Tree Physiol 33:845–854. https://doi.org/10.1093/treephys/tpt063
Singh A, Bajpai V, Kumar S, Sharma KR, Kumar B (2016) Profiling of gallic and ellagic acid derivatives in different plant parts of Terminalia arjuna by HPLC-ESI-QTOF-MS/MS. Nat Prod Commun 11:239–244. https://doi.org/10.1177/1934578x1601100227
Sobeh M, Mahmoud MF, Hasan RA, Abdelfattah MAO, Osman S, Rashid HO, El-Shazly AM, Wink M (2019) Chemical composition, antioxidant and hepatoprotective activities of methanol extracts from leaves of Terminalia bellirica and Terminalia sericea (Combretaceae). PeerJ 7:e6322. https://doi.org/10.7717/peerj.6322
Subramanian S, Graham MY, Yu O, Graham TL (2005) RNA interference of soybean isoflavone synthase genes leads to silencing in tissues distal to the transformation site and to enhanced susceptibility to Phytophthora sojae. Plant Physiol 137:1345–1353. https://doi.org/10.1104/pp.104.057257
Sun L, Tao S, Zhang S (2019) Characterization and quantification of polyphenols and triterpenoids in thinned young fruits of ten pear varieties by UPLC-Q TRAP-MS/MS. Molecules 24:159. https://doi.org/10.3390/molecules24010159
Tavares L, Fortalezas S, Carrilho C, McDougall GJ, Stewart D, Ferreira RB, Santos CN (2010) Antioxidant and antiproliferative properties of strawberry tree tissues. J Berry Res 1:3–12. https://doi.org/10.3233/BR-2010-001
Thomma BPHJ (2003) Alternaria spp.: from general saprophyte to specific parasite. Mol Plant Pathol 4:225–236. https://doi.org/10.1046/j.1364-3703.2003.00173.x
Torres JA, Valle F, Pinto C, García-Fuentes A, Salazar C, Cano E (2002) Arbutus unedo L. communities in southern Iberian Peninsula mountains. Plant Ecol 160:207–223. https://doi.org/10.1023/A:1015864821706
Trevors JT, Basaraba J (1980) Toxicity of benzoquinone and hydroquinone in short-term bacterial bioassays. Bull Environ Contam Toxicol 25:672–675. https://doi.org/10.1007/BF01985590
Tuominen A (2017) Tannins and other polyphenols in Geranium sylvaticum: identification, intraplant distribution and biological activity. University of Turku
Vargas P, Farias GA, Nogales J, Prada H, Carvajal V, Barón M, Rivilla R, Martín M, Olmedilla A, Gallegos MT (2013) Plant flavonoids target Pseudomonas syringae pv. tomato DC3000 flagella and type III secretion system. Environ Microbiol Rep 5:841–850. https://doi.org/10.1111/1758-2229.12086
Velasco P, Lema M, Francisco M, Soengas P, Cartea M (2013) In vivo and in vitro effects of secondary metabolites against Xanthomonas campestris pv. campestris. Molecules 18:11131–11143. https://doi.org/10.3390/molecules180911131
Verma N, Shukla S (2015) Impact of various factors responsible for fluctuation in plant secondary metabolites. J Appl Res Med Aromat Plants 2:105–113. https://doi.org/10.1016/j.jarmap.2015.09.002
Vu VQ (2011) ggbiplot: a ggplot2 based biplot. R package version 0.55. http://github.com/vqv/ggbiplot
Wickham H (2016) ggplot2: elegant graphics for data analysis. Springer-Verlag New York. ISBN 978–3–319–24277–4. https://ggplot2.tidyverse.org
Witzell J, Martín JA (2008) Phenolic metabolites in the resistance of northern forest trees to pathogens-past experiences and future prospects. Can J For Res 38:2711–2727. https://doi.org/10.1139/X08-112
Wrona M, Blasco S, Becerril R, Nerin C, Sales E, Asensio E (2019) Antioxidant and antimicrobial markers by UPLC–ESI-Q-TOF-MSE of a new multilayer active packaging based on Arctostaphylos uva-ursi. Talanta 196:498–509. https://doi.org/10.1016/j.talanta.2018.12.057
Wyrepkowski CC, Da Costa DLMG, Sinhorin AP, Vilegas W, De Grandis RA, Resende FA, Varanda EA, Dos Santos LC (2014) Characterization and quantification of the compounds of the ethanolic extract from Caesalpinia ferrea stem bark and evaluation of their mutagenic activity. Molecules 19:16039–16057. https://doi.org/10.3390/molecules191016039
Xia L, Xu C, Huang K, Lu J, Zhang Y (2019) Evaluation of phenolic compounds, antioxidant and antiproliferative activities of 31 grape cultivars with different genotypes. J Food Biochem 43:e12626. https://doi.org/10.1111/jfbc.12626
Xu W-H, Liang Q, Zhang Y-J, Zhao P (2015) Naturally occurring arbutin derivatives and their bioactivities. Chem Biodivers 12:54–81. https://doi.org/10.1002/cbdv.201300269
Yang L, Wen KS, Ruan X, Zhao YX, Wei F, Wang Q (2018) Response of plant secondary metabolites to environmental factors. Molecules 23:1–26. https://doi.org/10.3390/molecules23040762
Zengin G, Diuzheva A, Jekő J, Cziáky Z, Bulut G, Dogan A, Haznedaroglu MZ, Rengasamy KRR, Lobine D, Bahadori MB, Mahomoodally MF (2018) HPLC–MS/MS-based metabolic profiling and pharmacological properties of extracts and infusion obtained from Amelanchier parviflora var. dentata. Ind Crops Prod 124:699–706. https://doi.org/10.1016/j.indcrop.2018.08.042
Zhang Y, Xiong H, Xu X, Xue X, Liu M, Xu S, Liu H, Gao Y, Zhang H, Li X (2018) Compounds identification in semen cuscutae by ultra-high-performance liquid chromatography (UPLCs) coupled to electrospray ionization mass spectrometry. Molecules 23:1199. https://doi.org/10.3390/molecules23051199
Zhao P, Tanaka T, Hirabayashi K, Zhang Y-J, Yang C-R, Kouno I (2008) Caffeoyl arbutin and related compounds from the buds of Vaccinium dunalianum. Phytochemistry 69:3087–3094. https://doi.org/10.1016/j.phytochem.2008.06.001
Ziani BEC, Calhelha RC, Barreira JCM, Barros L, Hazzit M, Ferreira ICFR (2015) Bioactive properties of medicinal plants from the Algerian flora: Selecting the species with the highest potential in view of application purposes. Ind Crops Prod 77:582–589. https://doi.org/10.1016/j.indcrop.2015.09.034
Acknowledgements
Foundation for Science and Technology (Portugal) supported J. Martins PhD fellowship (SFRH/BD/122478/2016). The authors also acknowledge CESAM within PT2020 Partnership Agreement and Compete 2020 (UIDP/50017/2020 + UIDB/50017/2020), ReNATURE project (CENTRO-01–0145-FEDER-000007), Foundation for Science and Technology for the project nº 22125 “National Mass Spectrometry Network” (RNEM) of Portugal and Laboratory of Mass Spectrometry (LEM) of UC Node integrated in the RNEM, for MS analyses and project 0377_IBERPHENOL_6_E by Programa de Cooperación Interreg V-A España-Portugal (POCTEP) 2014–2020.
Author information
Authors and Affiliations
Contributions
JM: conceptualization, laboratory work, data analysis, manuscript preparation. TB: conceptualization, laboratory work, data analysis, manuscript revision and edition, funding. GP: manuscript revision and edition. JC: conceptualization, manuscript revision and edition, funding.
Corresponding author
Additional information
Communicated by LeBoldus.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Martins, J., Batista, T., Pinto, G. et al. Seasonal variation of phenolic compounds in Strawberry tree (Arbutus unedo L.) leaves and inhibitory potential on Phytophthora cinnamomi. Trees 35, 1571–1586 (2021). https://doi.org/10.1007/s00468-021-02137-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00468-021-02137-4