SpringerLink
Log in

Enzymatic activity assays and functional component profiling reveals coordinated defense response of mungbean against Cercospora canescens during leaf spot disease

  • Original Article
  • Published:
Plant Biotechnology Reports Aims and scope Submit manuscript

Abstract

Cercospora leaf spot (CLS) caused by Cercospora canescens is a formidable seed-borne disease and survives in crop debris or other host plants affecting mungbean cultivation. This study presents a comparative assessment of enzymatic activity assays and functional components in mungbean, focusing on the relationship of host plant and the pathogen, responsible for CLS. Comparative analyses were conducted between susceptible and resistant genotypes in both inoculated and uninoculated conditions to analyze defense-related activity like ascorbate peroxidase (APX), peroxidase (POD), lipid peroxidase (LPX), and catalase (CAT). In addition, C-based compounds including total phenols and flavonoids were analyzed through detached mungbean leaf method. The study further extended its analysis beyond leaves to other parts like pods, stems, and seeds. The host–pathogen relationship was investigated at various stages; control, 2 days after inoculation (DAI), 4 DAI, 6 DAI. At 6 DAI (T4), in the pods, the enzyme activity was 3.1-fold higher for ascorbate peroxidase, two-fold higher for peroxidase, and 1.8-fold higher for lipid peroxidase in case of susceptible genotype in contrast to mock inoculation (T1). For resistant genotype, the antioxidant activity at 6 DAI (T4) was 3.7-fold higher for ascorbate peroxidase, 2.73-fold higher for peroxidase, 1.8-fold higher for lipid peroxidase, and 2.7-fold higher for catalase compared to mock inoculation (T1). In pods, catalase activity was upregulated by 1.47-fold in resistant genotype and conversely, its activity was downregulated by 1.30-fold in pods of susceptible genotype at 6 DAI. Results indicate a positive regulation of catalase in pods of resistant genotype throughout infection period. These findings underscore the plant’s ability to mount a well-coordinated defense response, involving multiple components of the anti-oxidative system. The sophisticated regulation of these defense mechanisms reflects the adaptability and resilience of mungbean plants against pathogen challenges.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Germany)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Data availability

Additional data can be made available on request to corresponding author.

References

  • Abhay KP, Rishi RB, Lawrence K, Ramakrishnan MN (2018) Perspectives and challenges for sustainable management of fungal diseases of mungbean [Vigna radiata (L.) R. Wilczek var. radiata]: a review. Front Environ Sci 6:53

    Article  Google Scholar 

  • Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126

    Article  CAS  PubMed  Google Scholar 

  • Ammendolia DA, Bement WM, Brumell JH (2021) Plasma membrane integrity: implications for health and disease. BMC Biol 19:1–29

    Article  Google Scholar 

  • Andersen EJ, Ali S, Byamukama E, Yen Y, Nepal MP (2018) Disease resistance mechanisms in plants. Genes 9(7):339

    Article  PubMed  PubMed Central  Google Scholar 

  • Ayala A, Munoz MF, Argüelles S (2014) Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxid Med Cell Longev 2014:31

    Article  Google Scholar 

  • Basandrai AK, Pandey AK, Somta P, Basandrai D (2021) Macrophomina phaseolina-host interface: insights into an emerging dry rot pathogen of mungbean and urdbean, and its mitigation strategies. Plant Pathol 70:1263–1275

    Article  Google Scholar 

  • Basavarajappa DS, Kumar RS, Rudrappa M, Hiremath H, Almansour AI, Perumal K, Kantli GB, Nayaka S (2023) Acceleration of histopathological and biochemical defense patterns in mungbean with biopriming of Amycolatopsis sp. SND-1 against Cercospora leaf spot disease. J King Saud Univ Sci 35(8):102903

    Article  Google Scholar 

  • Batzer JC, Singh A, Rairdin A, Chiteri K, Mueller DS (2022) Mungbean: a preview of disease management challenges for an alternative US cash crop. J Integr Pest Manag 13(1):4

    Article  Google Scholar 

  • Bharti M, Chandra R, Kumar A, Kumar R, Yadav V, Yadav P (2017) Enzymatic response of mungbean (Vigna radiata) genotypes against Cercospora leaf spot disease. Indian J Agric Sci 87(7):930–933

    CAS  Google Scholar 

  • Bolwell GP, Wojtaszek P (2002) Mechanisms for the generation of reactive oxygen species in plant defence–a broad perspective. Physiol Mol Plant Pathol 51(6):347–366

    Article  Google Scholar 

  • Caverzan A, Passaia G, Rosa SB, Ribeiro CW, Lazzarotto F, Margis-Pinheiro M (2012) Plant responses to stresses: role of ascorbate peroxidase in the antioxidant protection. Genet Mol Biol 35:1011–1019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chand R, Pal C, Singh V, Kumar M, Singh VK, Chowdappa P (2015) Draft genome sequence of Cercospora canescens: a leaf spot causing pathogen. Curr Sci 10:2103–2110

    Article  Google Scholar 

  • Chandra S, Khan S, Avula B, Lata H, Yang MH, El Sohly MA, Khan IA (2014) Assessment of total phenolic and flavonoid content, antioxidant properties, and yield of aeroponically and conventionally grown leafy vegetables and fruit crops: a comparative study. Evid Based Complement Altern Med 2014:253875

    Article  Google Scholar 

  • Chang CC, Yang MH, Wen HM, Chern JC (2002) Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 10(3):3

    Google Scholar 

  • Cruz JE, Saldanha HC, Nascimento AM, Borges RB, Gomes MD, Freitas GR, Leal CM, Ferreira EA, da Silva Filho AA, Morais ER (2023) Evaluation of the antioxidant, antimicrobial, and anti-biofilm effects of the stem bark, leaf, and seed extracts from Hymenaea courbaril and characterization by UPLC-ESI-QTOF-MS/MS analysis. Antibiotics 12(11):1601

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dahiya PK, Linnemann AR, Van Boekel MA, Khetarpaul N, Grewal RB, Nout MJ (2015) Mung bean: technological and nutritional potential. Crit Rev Food Sci Nutr 55(5):670–688

    Article  CAS  PubMed  Google Scholar 

  • Debona D, Rodrigues FÁ, Rios JA, Nascimento KJ (2012) Biochemical changes in the leaves of wheat plants infected by Pyricularia oryzae. Phytopathology 102(12):1121–1129

    Article  CAS  PubMed  Google Scholar 

  • Djeridane A, Yousfi M, Nadjemi B, Boutassouna D, Stocker P, Vidal N (2006) Antioxidant activity of some Algerian medicinal plants extracts containing phenolic compounds. Food Chem 97(4):654–660

    Article  CAS  Google Scholar 

  • Du Y, Amin N, Ahmad N, Zhang H, Zhang Y, Song Y, Fan S, Wang P (2023) Identification of the function of the pathogenesis-related protein GmPR1L in the resistance of soybean to Cercospora so**a hara. Genes 14(4):920

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Duan C, Yu J, Bai J, Zhu Z, Wang X (2014) Induced defense responses in rice plants against small brown plant hopper infestation. Crop J 2(1):55–62

    Article  Google Scholar 

  • El-Khallal SM (2007) Induction and modulation of resistance in tomato plants against Fusarium wilt disease by bioagent fungi (arbuscular mycorrhiza) and/or hormonal elicitors (jasmonic acid and salicylic acid): 1-changes in growth, some metabolic activities and endogenous hormones related to defence mechanism. Aust J Basic Appl Sci 1(4):691–705

    CAS  Google Scholar 

  • Fahnenstich H, Scarpeci TE, Valle EM, Flugge UI, Maurino VG (2008) Generation of hydrogen peroxide in chloroplasts of Arabidopsis overexpressing glycolate oxidase as an inducible system to study oxidative stress. Plant Physiol 148(2):719–729

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Farahani AS, Taghavi M (2016) Changes of antioxidant enzymes of mungbean [Vigna radiata (L.) R. Wilczek] in response to host and non-host bacterial pathogens. J Plant Prot Res 56:1

    Article  Google Scholar 

  • Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17(7):1866–1875

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gaschler MM, Stockwell BR (2017) Lipid peroxidation in cell death. Biochem Biophys Res Commun 482(3):419–425

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hameed A, Bibi N, Akhter J, Iqbal N (2011) Differential changes in antioxidants, proteases, and lipid peroxidation in flag leaves of wheat genotypes under different levels of water deficit conditions. Plant Physiol Biochem 49(2):178–185

    Article  CAS  PubMed  Google Scholar 

  • Huang H, Ullah F, Zhou DX, Yi M, Zhao Y (2019) Mechanisms of ROS regulation of plant development and stress responses. Front Plant Sci 10:800

    Article  PubMed  PubMed Central  Google Scholar 

  • Ishihara A (2021) Defense mechanisms involving secondary metabolism in the grass family. J Pestic Sci 46(4):382–392

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kaur K, Grewal SK, Singh S, Rani U, Bhardwaj RD (2021) Timing and intensity of upregulated defensive enzymes is a key factor determining resistance in chickpea to Ascochyta rabiei. Physiol Mol Plant Pathol 114:101645

    Article  CAS  Google Scholar 

  • Kaur S, Samota MK, Choudhary M, Choudhary M, Pandey AK, Sharma A, Thakur J (2022) How do plants defend themselves against pathogens-biochemical mechanisms and genetic interventions. Physiol Mol Biol Plants 28(2):485–504

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Latef AA, Chaoxing H (2011) Effect of arbuscular mycorrhizal fungi on growth, mineral nutrition, antioxidant enzymes activity and fruit yield of tomato grown under salinity stress. Sci Hortic 127(3):228–233

    Article  Google Scholar 

  • Lavanya GR, Srivastava J, Ranade SA (2008) Molecular assessment of genetic diversity in mungbean germplasm. J Genet 87:65–74

    Article  CAS  PubMed  Google Scholar 

  • Li T, Bao Y, Qiu H, Tong W (2021) Boosted peroxidase-like activity of metal-organic framework nanoparticles with single atom Fe (III) sites at low substrate concentration. Anal Chim Acta 1152:338299

    Article  CAS  PubMed  Google Scholar 

  • Lin CC, Kao CH (2001) Cell wall peroxidase activity, hydrogen peroxide level and NaCl-inhibited root growth of rice seedlings. Plant Soil 230:135-143.

    Article  CAS  Google Scholar 

  • Lismont C, Revenco I, Fransen M (2019) Peroxisomal hydrogen peroxide metabolism and signaling in health and disease. Int J Mol Sci 20(15):3673

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lubaina AS, Murugan K (2013) Ultrastructural changes and oxidative stress markers in wild and cultivar Sesamum orientale L. following alternaria sesami (Kawamura) Mohanty and Behera. inoculation. Indian J Exp Biol 51(8):670–680

    CAS  PubMed  Google Scholar 

  • Mahapatra SS, Swain D, Beura SK, Rout GR (2022) Identification of mungbean germplasm for resistance against Cercospora canescens and to study the association of biochemical parameters with defense mechanisms. Agron J 114(2):1184–1199

    Article  CAS  Google Scholar 

  • Mansoor S, Ali Wani O, Lone JK, Manhas S, Kour N, Alam P, Ahmad A, Ahmad P (2022) Reactive oxygen species in plants: from source to sink. Antioxidants 11(2):225

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mansoor S, Ali A, Kour N, Bornhorst J, AlHarbi K, Rinklebe J, Chung YS (2023) Heavy metal induced oxidative stress mitigation and ROS scavenging in plants. Plants 12(16):3003

  • Nair RM, Pandey AK, War AR, Hanumantharao B, Shwe T, Alam AK, Pratap A, Malik SR, Karimi R, Mbeyagala EK, Douglas CA (2019) Biotic and abiotic constraints in mungbean production—progress in genetic improvement. Front Plant Sci 10:1340

    Article  PubMed  PubMed Central  Google Scholar 

  • Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22(5):867–880

    CAS  Google Scholar 

  • Nandi A, Yan LJ, Jana CK, Das N (2019) Role of catalase in oxidative stress-and age-associated degenerative diseases. Oxid Med Cell Longev 11:2019

    Google Scholar 

  • Nasser Sewelam Peer MS (2016) Global plant stress signaling: reactive oxygen species at the cross-road. Front Plant Sci 1152:338299

    Google Scholar 

  • Noctor G, Foyer CH (2016) Intracellular redox compartmentation and ROS-related communication in regulation and signaling. Plant Physiol 171:1581–1592

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sachdev S, Ansari SA, Ansari MI, Fujita M, Hasanuzzaman M (2021) Abiotic stress and reactive oxygen species: generation, signaling, and defense mechanisms. Antioxidants 10(2):277

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Saenen E, Horemans N, Vanhoudt N, Vandenhove H, Biermans G, van Hees M, Wannijn J, Vangronsveld J, Cuypers A (2015) Oxidative stress responses induced by uranium exposure at low pH in leaves of Arabidopsis thaliana plants. J Environ Radioact 150:36–43

    Article  CAS  PubMed  Google Scholar 

  • Sala, J. M., & Lafuente, M. T. (2000). Catalase enzyme activity is related to tolerance of mandarin fruits to chilling. Postharvest Biol Tech 20(1):81-89.

    Article  CAS  Google Scholar 

  • Scandalios JG (2005) Oxidative stress: molecular perception and transduction of signals triggering antioxidant gene defenses. Braz J Med Biol Res 38:995–1014

    Article  CAS  PubMed  Google Scholar 

  • Sharma P, Jha AB, Dubey RS, Pessarakli M (2012) Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. J Bot 2012:1–26

    Article  Google Scholar 

  • Siddique Z, Akhtar KP, Hameed A, Sarwar N, Imran-Ul-Haq KSA (2014) Biochemical alterations in leaves of resistant and susceptible cotton genotypes infected systemically by cotton leaf curl Burewala virus. J Plant Interact 9(1):702–711

    Article  Google Scholar 

  • Singh M, Avtar R, Pal A, Punia R, Singh VK, Bishnoi M, Singh A, Choudhary RR, Mandhania S (2020) Genotype-specific antioxidant responses and assessment of resistance against Sclerotinia sclerotiorum causing Sclerotinia rot in Indian mustard. Pathogens 9(11):892

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sousa RH, Carvalho FE, Lima-Melo Y, Alencar VT, Daloso DM, Margis-Pinheiro M, Komatsu S, Silveira JA (2019) Impairment of peroxisomal APX and CAT activities increases protection of photosynthesis under oxidative stress. J Exp Bot 70(2):627–639

    Article  CAS  PubMed  Google Scholar 

  • Su LJ, Zhang JH, Gomez H, Murugan R, Hong X, Xu D, Jiang F, Peng ZY (2019) Reactive oxygen species-induced lipid peroxidation in apoptosis, autophagy, and ferroptosis. Oxid Med Cell Longev 2019:5080843

    Article  PubMed  PubMed Central  Google Scholar 

  • Tripathy BC, Oelmüller R (2012) Reactive oxygen species generation and signaling in plants. Plant Signal Behav 7:1621–1633

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yang H, Zhou M, Li H, Wei T, Tang C, Zhou Y, Long X (2020) Effects of low-level lipid peroxidation on the permeability of nitro-aromatic molecules across a membrane: a computational study. ACS Omega 5(10):4798–4806

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yanti Y (2015) Peroxidase enzyme activity of rhizobacteria-introduced shallots bulbs to induce resistance of shallot towards bacterial leaf blight (Xanthomonas axonopodis pv allii). Procedia Chem 14:501–507

    Article  CAS  Google Scholar 

  • Zandi P, Schnug E (2022) Reactive oxygen species, antioxidant responses and implications from a microbial modulation perspective. Biology 11(2):155

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zeeshan M, Murugadas A, Ghaskadbi S, Rajendran RB, Akbarsha MA (2014) ROS dependent copper toxicity in Hydra-biochemical and molecular study. Comp Biochem Physiol C Toxicol Pharmacol 185:1–2

    Google Scholar 

  • Zhan J, Thrall PH, Burdon JJ (2014) Achieving sustainable plant disease management through evolutionary principles. Trends Plant Sci 19(9):570–575

    Article  CAS  PubMed  Google Scholar 

  • Zheng HZ, Kim YW, Lee HJ, Park RD, Jung WJ, Kim YC, Lee SH, Kim TH, Kim KY (2004) Quantitative changes of PR proteins and antioxidative enzymes in response to Glomus intraradices and Phytophthora capsici in pepper (Capsicum annuum L.) plants. J Microbiol Biotechnol 14(3):553–562

    CAS  Google Scholar 

  • Zhu Yi Shen ZY, Sun Shuai SS, Fitzgerald R (2014) Mungbean proteins and peptides: nutritional, functional and bioactive properties. Food Nutr Res 62:1290

    Google Scholar 

  • Zieslin N, Ben-Zaken RJ (1993) Peroxidase activity and presence of phenolic substances in peduncles of rose flowers. Plant Physiol Biochem 31:333–339

    CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF), funded by the Ministry of Education (2016R1A6A1A03012862).

Author information

Authors and Affiliations

  1. Division of Genetics and Plant Breeding, Faculty of Agriculture, SKUAST-Kashmir, Wadura, 193201, India

    Mohammad Irfan & Mohd Ashraf Bhat

  2. Division of Plant Pathology, Faculty of Agriculture, SKUAST-Kashmir, Wadura, 193201, India

    Uzma Rashid

  3. Department of Biological Sciences, Faculty of Science, King Abdulaziz University, 21589, Jeddah, Saudi Arabia

    Enas A. Almanzalawi & Tahani M. Alqahtani

  4. Subtropical/Tropical Organism Gene Bank, Jeju National University, Jeju, 63243, Republic of Korea

    Sheikh Mansoor & Kyung-Hwan Boo

  5. Department of Biotechnology, College of Applied Life Science (SARI), Jeju National University, Jeju, 63243, Republic of Korea

    Kyung-Hwan Boo

Authors
  1. Mohammad Irfan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mohd Ashraf Bhat
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Uzma Rashid
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Enas A. Almanzalawi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tahani M. Alqahtani
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sheikh Mansoor
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kyung-Hwan Boo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Mohd Ashraf Bhat or Kyung-Hwan Boo.

Ethics declarations

Conflict of interest

The authors declare no conflict of interest.

Ethical approval

Not applicable.

Consent to participate

Not applicable.

Consent to publication

Not applicable.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOC 96 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Irfan, M., Bhat, M.A., Rashid, U. et al. Enzymatic activity assays and functional component profiling reveals coordinated defense response of mungbean against Cercospora canescens during leaf spot disease. Plant Biotechnol Rep (2024). https://doi.org/10.1007/s11816-024-00918-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11816-024-00918-0

Keywords

Search

Navigation