Acta Physiologiae Plantarum

, 41:186 | Cite as

Physiological and biochemical responses of soybean to white mold affected by manganese phosphite and fluazinam

  • M. I. C. Novaes
  • D. Debona
  • I. R. F. Fagundes-Nacarath
  • V. V. Brás
  • F. A. RodriguesEmail author
Original Article


White mold, caused by Sclerotinia sclerotiorum, is one of the most important diseases affecting soybean and its control has been difficult to achieve. This study aimed to investigate the potential of manganese (Mn) phosphite and fluazinam in protecting soybean plants against S. sclerotiorum infection by examining the photosynthetic performance (leaf gas exchange and chlorophyll (Chl) a fluorescence parameters), activities of defense enzymes [chitinase (CHI), β-1,3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), and polyphenol oxidase (PPO)] as well as those related to the antioxidant metabolism (superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), and ascorbate peroxidase (APX)) and the concentrations of hydrogen peroxide (H2O2), superoxide (O2), and malondialdehyde (MDA). White mold development was completely inhibited by fluazinam. Soybean metabolism was not changed by fluazinam. White mold severity was significantly reduced on plants sprayed with Mn phosphite, which showed a better photosynthetic performance than the non-sprayed plants. Mycelial growth of S. sclerotiorum was inhibited by Mn phosphite. Activities of CAT, POX, and SOD decreased while CHI, GLU, and PAL activities increased at 96 hai for Mn phosphite-sprayed plants compared to non-sprayed plants. In conclusion, Mn phosphite affected white mold development and pathogen-induced physiological impairments in soybean leaflets due to its dual mode of action.


Glycine max Sclerotinia sclerotiorum Antioxidant enzymes Defense enzymes Photosynthesis 



Prof. Fabrício A. Rodrigues thanks the “Conselho Nacional de Desenvolvimento Científico e Tecnológico” (CNPq) for his research fellowship. This study was supported by grants from CAPES, CNPq, and FAPEMIG to Prof. Rodrigues. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001.


  1. Araujo L, Valdebenito-Sanhueza RM, Stadnik MJ (2010) Avaliação de formulações de fosfito de potássio sobre Colletotrichum gloeosporioides in vitro e no controle pós-infeccional da mancha foliar de Glomerella em macieira. Trop Plant Pathol 35:54–59Google Scholar
  2. Araujo L, Bispo WMS, Rios VS, Fernandes SA, Rodrigues FA (2015) Induction of the phenylpropanoid pathway by acibenzolar-S-methyl and potassium phosphite increases mango resistance to Ceratocystis fimbriata infection. Plant Dis 99:447–459PubMedGoogle Scholar
  3. Bae YS, Knudsen GR (2007) Effect of sclerotial distribution pattern of Sclerotinia sclerotiorum on biocontrol efficacy of Trichoderma harzianum. Appl Soil Ecol 35:21–24Google Scholar
  4. Bateman DF, Beer SV (1965) Simultaneous production and synergistic action of oxalic acid and polygalacturonase during pathogenesis by Sclerotium rolfsii. Phytopathology 55:204–211PubMedGoogle Scholar
  5. Bermúdez-Cardona MB, Wordell Filho JA, Rodrigues FA (2015) Leaf gas exchange and chlorophyll a fluorescence in maize leaves infected with Stenocarpella macrospora. Phytopathology 105:26–34PubMedGoogle Scholar
  6. Bock CH, Brenneman TB, Hotchkiss MW, Wood BW (2013) Evaluation of a phosphite fungicide to control pecan scab in the southeastern USA. Crop Prot 36:58–64Google Scholar
  7. Boland GJ, Hall R (1987) Evaluating soybeans cultivars for resistance to Sclerotinia sclerotiorum under field conditions. Plant Dis 71:934–936Google Scholar
  8. Brackmann A, Giehl RFH, Sestari I, Steffens CA (2004) Fosfitos para o controle de podridões pós-colheita em maçãs ‘Fuji’ durante o armazenamento refrigerado. Cienc Rural 34:1039–1042Google Scholar
  9. Bradford MN (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedPubMedCentralGoogle Scholar
  10. Bu JW, Yao G, Gao HY, Jia YJ, Zhang LT, Cheng DD, Wang X (2009) Inhibition mechanism of photosynthesis in cucumber leaves infected by Sclerotinia sclerotiorum (Lib.) de Bary. Acta Phytopathol Sinc 39:613–621Google Scholar
  11. Cakmak I, Horst WJ (1991) Effect of aluminum on lipid peroxidation, superoxide dismutase, catalase, and peroxidase activities in root tips of soybean (Glycine max). Physiol Plant 83:463–468Google Scholar
  12. Carmona MA, Simonetti E, Ravotti ME, Scandiani MM, Luque AG, Formento NA, Sautua FJ (2017) In vitro antifungal/fungistatic activity of manganese phosphite against soybean soil-borne pathogens. Phyton Int J Exp Bot 85:265–269Google Scholar
  13. Cerqueira A, Alves A, Berenguer H, Correia B, Gómez-Cadenas A, Diez JJ, Monteiro P, Pinto G (2017) Phosphite shifts physiological and hormonal profile of Monterey pine and delays Fusarium circinatum progression. Plant Physiol Biochem 114:88–99PubMedGoogle Scholar
  14. Chaitanya KSK, Naithani SC (1994) Role of superoxide lipid peroxidation and superoxide dismutase in membrane perturbation during loss of viability in seeds of Shorea robusta Gaertn.f. New Phytol 126:623–627Google Scholar
  15. Christensen TH, Zhang Z, Wei Y, Collinge DB (1997) Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction. Plant J 11:1187–1194Google Scholar
  16. Clark RB (1975) Characterization of phosphates in intact maize roots. J Agric Food Chem 23:458–460PubMedGoogle Scholar
  17. Conrath U, Pieterse CMJ, Mauch-Mani B (2002) Priming in plant-pathogen interactions. Trends Plant Sci 7:210–216PubMedGoogle Scholar
  18. Costa BHG, Resende MLV, Monteiro ACA, Junior PMR, Botelho DMS, Silva BM (2018) Potassium phosphites in the protection of common bean plants against anthracnose and biochemical defence responses. J Phytopathol 166:95–102Google Scholar
  19. Dalio RJD, Fleischmann F, Humez M, Oswald W (2014) Phosphite protects Fagus sylvatica seedlings towards Phytophthora plurivora via local toxicity, priming and facilitation of pathogen recognition. PLoS One 9:e87860PubMedPubMedCentralGoogle Scholar
  20. Daniel R, Guest D (2006) Defense responses induced by potassium phosphonate in Phytophthora palmivora-challenged Arabidopsis thaliana. Physiol Mol Plant Pathol 67:194–201Google Scholar
  21. Debona D, Rodrigues FA, Rios JA, Martins SC, Pereira LF, DaMatta FM (2014) Limitations to photosynthesis in leaves of wheat plants infected by Pyricularia oryzae. Phytopathology 104:34–39PubMedGoogle Scholar
  22. Debona D, Nascimento KJT, Gomes JGO, Aucique-Pérez CE, Rodrigues FA (2016) Physiological changes promoted by a strobilurin fungicide in the rice-Bipolaris oryzae interaction. Pest Biochem Physiol 10:8–16Google Scholar
  23. Dianese AL, Blum LEB, Dutra JB, Lopes LF (2009) Aplicação de fosfito de potássio, cálcio ou magnésio para a redução da podridão-do-pé do mamoeiro em casa de vegetação. Cienc Rural 29:2309–2314Google Scholar
  24. Doke N, Miura Y, Sanchez LM, Park HJ, Noritake T, Yoshioka H, Kawakita K (1996) The oxidative burst protects plants against pathogen attack: mechanism and role as an emergency signal for plant biodefence. Gene 179:45–51PubMedGoogle Scholar
  25. Fagundes-Nacarath IRF, Debona D, Brás VV, Silveira PR, Rodrigues FA (2018) Phosphites attenuate Sclerotinia sclerotiorum-induced physiological impairments in common bean. Acta Physiol Plant 40:198Google Scholar
  26. Fehr WR, Caviness CE, Burmood DT, Pennington JS (1971) Stage of development descriptions for soybeans, Glycine max (L.) Merrill. Crop Sci 11:929–931Google Scholar
  27. Fortunato AA, Debona D, Bernadeli AMA, Rodrigues FA (2015a) Changes in the antioxidant system in soybean leaves infected by Corynespora cassiicola. Phytopathology 105:1050–1058PubMedGoogle Scholar
  28. Fortunato AA, Debona D, Bernardeli AMA, Rodrigues FA (2015b) Defence-related enzymes in soybean resistance to target spot. J Phytopathol 163:731–742Google Scholar
  29. Gay C, Gebicki JM (2000) A critical evaluation of the effect of sorbitol on the ferric-xylenol orange hydroperoxide assay. Anal Biochem 284:217–220Google Scholar
  30. Guimarães RL, Stotz HU (2004) Oxalate production by Sclerotinia sclerotiorum deregulates guard cells during infection. Plant Physiol 136:3703–3711PubMedPubMedCentralGoogle Scholar
  31. Guo X, Stotz H (2010) ABA signaling inhibits oxalate-induced production of reactive oxygen species and protects against Sclerotinia sclerotiorum in Arabidopsis thaliana. Eur J Plant Pathol 128:7–19Google Scholar
  32. Guo Z, Miyoshi H, Kimyoji T, Haga T, Fujita T (1991) Uncoupling activity of a newly developed fungicide, fluazinam [3-chloro-N-(3-chloro-2,6-dinitro-4-trifluoromethylphenyl)-5-trifluoromethyl-2-pyridinamine]. Biochim Biophys Acta Bioenerg 1056:89–92Google Scholar
  33. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplast. I. Kinetics and stoichometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198PubMedPubMedCentralGoogle Scholar
  34. Heffer Link V, Johnson KB (2007) White mold: the plant health instructor. Accessed 9 June 2018
  35. Hegedus DD, Rimmer SR (2005) Sclerotinia sclerotiorum: when to be or not to be a pathogen? FEMS Microbiol Lett 251:177–184PubMedGoogle Scholar
  36. Kim HS, Diers BW (2014) Inheritance of partial resistance to Sclerotinia stem rot in soybean. Crop Sci 40:55–61Google Scholar
  37. Klughammer C, Schreiber U (2008) Complementary PSII quantum yield calculated from simple fluorescence parameters measured by PAM fluorometry and saturation pulse method. PAM Appl Notes 1:27–35Google Scholar
  38. Lehner MS, Paula Junior TJ, Silva RA, Vieira RF, Carneiro JES, Schnabel G, Mizubuti ESG (2015) Fungicide sensitivity of Sclerotinia sclerotiorum: a thorough assessment using discriminatory dose, EC50, high-resolution melting analysis, and description of new point mutation associated with thiophanate-methyl resistance. Plant Dis 99:1537–1543PubMedGoogle Scholar
  39. Lehner MS, Pethybridge SJ, Meyer MC, Del Ponte EM (2017) Meta-analytic modelling of the incidence-yield and incidence-sclerotial production relationships in soybean white mould epidemics. Plant Pathol 66:460–468Google Scholar
  40. Lobato MC, Olivieri FP, Daleo GR, Andreu AB (2010) Antimicrobial activity of phosphites against different potato pathogens. J Plant Dis Protec 117:102–109Google Scholar
  41. Machinandiarena MF, Lobato MC, Feldman ML, Daleo GR, Andreu AB (2012) Potassium phosphite primes defense responses in potato against Phytophthora infestans. J Plant Physiol 149:1417–1424Google Scholar
  42. Malenčić DL, Kiprovski B, Popović M, Prvulović D, Miladinović J, Djordjević V (2010) Changes in antioxidant systems in soybean as affected by Sclerotinia sclerotiorum (Lib.) de Bary. Plant Physiol Biochem 48:903–908PubMedGoogle Scholar
  43. Mandal S, Mitra A, Mallick N (2008) Biochemical characterization of oxidative burst during interaction between Solanum lycopersicum and Fusarium oxysporum f. sp. lycopersici. Physiol Mol Plant Pathol 72:56–61Google Scholar
  44. McCrearya CM, Depuydta D, Vynb RJ, Gillars CL (2016) Fungicide efficacy of dry bean white mold [Sclerotinia sclerotiorum(Lib.) de Bary, causal organism] and economic analysis at moderate to high disease pressure. Crop Prot 82:75–81Google Scholar
  45. Meyer MC, Campos HD, Godoy CV, Utiamada CM (2014) Ensaios cooperativos de controle químico de mofo branco na cultura da soja: safras 2009 a 2012. Accessed 15 July 2018
  46. Mueller DS, Dorrance AE, Derksen RC, Ozkan E, Kurle JE, Grau CR, Gaska JM, Hartman GL, Bradley CA, Pedersen WL (2002) Efficacy of fungicides on Sclerotinia sclerotiorum and their potential for control of Sclerotinia stem rot on soybean. Plant Dis 86:26–31PubMedGoogle Scholar
  47. Mueller DS, Bradley C, Chilvers M, Esker P, Malvick D, Peltier A, Sisson A, Wise K (2015) White mold: Soybean Disease Management. Crop Protection Network, 1005: [ Accessed 10 June 2018
  48. Nascimento KJT, Araujo L, Resende RS, Schurt DA, Silva WL, Rodrigues FA (2016) Silicon, acibenzolar-S-methyl and potassium phosphite in the control of brown spot in rice. Bragantia 75:212–221Google Scholar
  49. Nojosa GBA, Resende MLV, Barguil BM, Moraes SRG, Vilas Boas CH (2009) Efeito de indutores de resistência em cafeeiro contra a mancha de Phoma. Summa Phytopathol 35:60–62Google Scholar
  50. Panicker S, Gangadharam K (1999) Controlling downy mildew of maize caused by Peronosclerospora sorghi by foliar sprays of phosphonic acid compounds. Crop Protec 18:115–118Google Scholar
  51. Peruch LAM, Brunna ED (2008) Relação entre doses de calda bordalesa e de fosfito potássio na intensidade do míldio e na produtividade da videira cv. ‘Goethe’. Cienc Rural 38:2413–2418Google Scholar
  52. Perveen K, Haseen A, Shukla PK (2010) Effect of Sclerotinia sclerotiorum on the disease development, growth, oil yield and biochemical changes in plants of Mentha arvensis. Saudi J Biol Sci 17:291–294PubMedPubMedCentralGoogle Scholar
  53. Pinto KMS, Nascimento LC, Gomes ECS, Silva HF, Miranda JR (2012) Efficiency of resistance elicitors in the management of grapevine downy mildew Plasmopara viticola: epidemiological, biochemical and economic aspects. Eur J Plant Pathol 134:745–754Google Scholar
  54. Rios JA, Rios VS, Aucique-Pérez CE, Cruz MFA, Morais LE, DaMatta FM, Rodrigues FA (2017) The photosynthetic performance and source-sink relationships are altered on wheat plants infected by Pyricularia oryzae. Plant Pathol 66:1496–1507Google Scholar
  55. Ryals J, Uknes S, Wars E (1994) Systemic acquired resistance. Plant Physiol 104:1109–1112PubMedPubMedCentralGoogle Scholar
  56. Simonetti E, Viso NP, Montecchia M, Zilli C, Balestrasse K, Carmona M (2015) Evaluation of native bacteria and manganese phosphite for alternative control of charcoal root rot of soybean. Microbiol Res 180:40–48PubMedGoogle Scholar
  57. Smillie R, Grant BR, Guest D (1989) The mode of action of phosphite: evidence for both direct and indirect modes of action on three Phytophthora spp. in plants. Phytopathology 79:921–926Google Scholar
  58. Sumida CH, Canteri MG, Peitil DC, Tibolla F, Orsini IP, Araujo FA, Chagas DF, Calvos NS (2015) Chemical and biological control of Sclerotinia stem rot in the soybeans crop. Cienc Rural 45:760–766Google Scholar
  59. Tariq VN, Jeffries P (1985) Changes occurring in chloroplasts of Phaseolus following infection by Sclerotinia: a cytochemical study. J Cell Sci 75:195–295PubMedGoogle Scholar
  60. Vallad GE, Goodman RM (2004) Systemic acquired resistance and induced systemic resistance in conventional agriculture. Crop Sci 44:1920–1934Google Scholar
  61. Van Loon LC, Rep M, Pieterse CMJ (2006) Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopathol 44:135–162PubMedGoogle Scholar
  62. Williams B, Kabbage M, Kim HJ, Britt R, Dickman MB (2011) Tipping the balance: Sclerotinia sclerotiorum secreted oxalic acid suppresses host defenses by manipulating the host redox environment. Plos Pathog 7:e1002107PubMedPubMedCentralGoogle Scholar
  63. Wellburn AR (1994) The spectral determination of chlorophylls a and b, as well as total carotenoids, using various solvents with spectrophotometers of different resolution. J Plant Physiol 144:307–313Google Scholar
  64. Xavier SA, Koga LJ, Barros DCM, Canteri MG, Lopes ION, Godoy CV (2015) Variação da sensibilidade de populações de Phakopsora pachyrhizi a fungicidas inibidores da desmetilação no Brasil. Summa Phytopathol 41:191–196Google Scholar
  65. Yang XB, Lundeen P, Uphoff MD (1999) Soybean varietal response and yield loss caused by Sclerotinia sclerotiorum. Plant Dis 83:456–461PubMedGoogle Scholar
  66. Yang C, Zhang Z, Gao H, Liu M, Fan X (2014) Mechanisms by which the infection of Sclerotinia sclerotiorum (Lib.) de Bary affects the photosynthetic performance in tobacco leaves. BMC Plant Biol 14:1–11Google Scholar
  67. Zhou J, Sun A, Xing D (2013) Modulation of cellular redox status by thiamine-activated NADPH oxidase confers Arabidopsis resistance to Sclerotinia sclerotiorum. J Exp Bot 64:3261–3272PubMedGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2019

Authors and Affiliations

  1. 1.Laboratório da Interação Planta-Patógeno, Departamento de FitopatologiaUniversidade Federal de ViçosaViçosaBrazil

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