Redox status regulation and action of extra- and intravascular defense mechanisms are associated with bean resistance against Fusarium oxysporum f. sp. phaseoli

Abstract

Genetic resistance is the main strategy to control one of the most destructive diseases of common bean (Phaseolus vulgaris L), i.e., the Fusarium wilt caused by Fusarium oxysporum f. sp. phaseoli (Fop). However, little is known on host defense reactions in Fop-bean interaction. Thus, this work examined the defense mechanisms in root and hypocotyl tissues of common bean against Fop. Resistant and susceptible bean plants were inoculated by dipping their roots in a conidial suspension. Cross sections of roots and hypocotyls were observed in light microscopy at 1, 3, 6, and 9 days after inoculation (dai) to monitor Fop colonization, and at 3 and 9 dai to detect callose, carbohydrates, lipids, phenolics, and protein, and under electronic microscopy at 9 dai to observe ultrastructural changes in xylem cells. The content of hydrogen peroxide (H2O2), lipid peroxidation, and activity of the antioxidant enzymes ascorbate peroxidase (EC 1.11.1.11) and catalase (EC 1.11.1.6) were monitored spectrophotometrically in roots and hypocotyls at 0, 1, 3, 6, and 9 dai. Fop colonized inter- and intracellularly the epidermis and cortex reaching the xylem vessels faster in susceptible genotype. Fop inoculation induced phenolics and carbohydrates accumulation, callose deposition, and formation of occlusion material inside xylem vessels mainly in resistant genotype. Lipid peroxidation occurred mainly in susceptible plants. In contrast, the antioxidant enzymes seem to have contributed to reducing damage caused by H2O2 accumulation in resistant plants. This study gives evidences that inter- and intracellular physicochemical mechanisms can act together to delay Fop colonization in resistant plants.

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Abbreviations

APX:

Ascorbate peroxidase

BHT:

Butylated hydroxytoluene

CAT:

Catalase

CBB:

Coomassie brilliant blue

CWA:

Cell wall apposition

dai:

Days after inoculation

Fop :

Fusarium oxysporum f. sp. phaseoli

FOX:

Ferrous oxidation xylenol orange

FW:

Fresh weight

Fw:

Fusarium wilt

H2O2 :

Hydrogen peroxide

LHPO:

Lipid hydroperoxide

MDA:

Malondialdehyde

ROS:

Reactive oxygen species

TBO:

Toluidine blue O

TCA:

Trichloroacetic acid

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Acknowledgments

We are grateful to the staff of the Multiuser Laboratory of Studies in Biology (LAMEB-UFSC), Central Laboratory of Electron Microscopy (LCME-UFSC), and Hydroponics Laboratory (LABHIDRO-UFSC) for their assistance, space, and equipment available in these laboratories. F.M.Q. is grateful for a master’s degree scholarship from CAPES/PROEX. M.J.S. is a research member of the National Council and Technological Development (CNPq).

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Correspondence to Felipe M. de Quadros or Marciel J. Stadnik.

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de Quadros, F.M., de Freitas, M.B., Simioni, C. et al. Redox status regulation and action of extra- and intravascular defense mechanisms are associated with bean resistance against Fusarium oxysporum f. sp. phaseoli. Protoplasma (2020). https://doi.org/10.1007/s00709-020-01521-0

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Keywords

  • Fusarium wilt
  • Disease resistance
  • Histochemical/fluorescence reactions
  • Cell ultrastructure
  • Defense mechanisms