Skip to main content

Advertisement

SpringerLink
Log in

Photosynthetic and biochemical mechanisms of an EMS-mutagenized cowpea associated with its resistance to cowpea severe mosaic virus

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

Abstract

Key message

The seed treatment of a CPSMV-susceptible cowpea genotype with the mutagenic agent EMS generated mutagenized resistant plantlets that respond to the virus challenge by activating biochemical and physiological defense mechanisms.

Abstract

Cowpea is an important crop that makes major nutritional contributions particularly to the diet of the poor population worldwide. However, its production is low, because cowpea is naturally exposed to several abiotic and biotic stresses, including viral agents. Cowpea severe mosaic virus (CPSMV) drastically affects cowpea grain production. This study was conducted to compare photosynthetic and biochemical parameters of a CPSMV-susceptible cowpea (CE-31 genotype) and its derived ethyl methanesulfonate-mutagenized resistant plantlets, both challenged with CPSMV, to shed light on the mechanisms of virus resistance. CPSMV inoculation was done in the fully expanded secondary leaves, 15 days after planting. At 7 days post-inoculation, in vivo photosynthetic parameters were measured and leaves collected for biochemical analysis. CPSMV-inoculated mutagenized-resistant cowpea plantlets (MCPI) maintained higher photosynthesis index, chlorophyll, and carotenoid contents in relation to the susceptible (CE-31) CPSMV-inoculated cowpea (CPI). Visually, the MCPI leaves did not exhibit any viral symptoms neither the presence of the virus as examined by RT-PCR. In addition, MCPI showed higher SOD, GPOX, chitinase, and phenylalanine ammonia lyase activities, H2O2, phenolic contents, and cell wall lignifications, but lower CAT and APX activities in comparison to CPI. All together, these photosynthetic and biochemical changes might have contributed for the CPSMS resistance of MCPI. Contrarily, CPI plantlets showed CPSMV accumulation, severe disease symptoms, reduction in the photosynthesis-related parameters, chlorophyll, carotenoid, phenolic compound, and H2O2 contents, in addition to increased β-1,3-glucanase, and catalase activities that might have favored viral infection.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Ainsworth EA, Gillespie KM (2007) Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin-Ciocalteu reagent. Nature 2:875–877

    CAS  Google Scholar 

  • Barna B, Fodor J, Harrach BD, Pogány M, Király Z (2012) The Janus face of reactive oxygen species in resistance and susceptibility of plants to necrotrophic and biotrophic pathogens. Plant Physiol Biochem 59:37–43

    Article  CAS  PubMed  Google Scholar 

  • Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287

    Article  CAS  PubMed  Google Scholar 

  • Boller T (1993) Biochemical analysis of chitinase and β-1,3-glucanases. In: Curr SJ (ed) Molecular plant pathology: a practical approach. Oxford University Press, New York, pp 23–29

    Google Scholar 

  • Bolton MD (2009) Primary metabolism and plant defense—fuel for the fire. Mol Plant Microbe Interact 224:87–97

    Google Scholar 

  • Booker HM, Umaharan P, McDavid CR (2005) Effect of Cowpea Severe Mosaic Virus on crop growth characteristic and yield of cowpea. Plant Dis 89:515–520

    Article  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of micro-gram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Brioso PST, Santiago LM, Anjos JRN, Oliveira DE (1996) Identification of species of the Comovirus genus by polymerase chain reaction. Fitopatol Brasil 21:219–225

    CAS  Google Scholar 

  • Chaerle L, Van Caeneghem W, Messens E, Lambers H, Van Montagu M, Van Der Straeten D (1999) Presymptomatic visualization of plant–virus interactions by thermography. Nat Biotechnol 17:813–816

    Article  CAS  PubMed  Google Scholar 

  • Chaman ME, Copaja SV, Argandona VH (2003) Relationships between salicylic acid content, phenylalanine ammonia-lyase (PAL) activity, and resistance of barley to aphid infestation. J Agric Food Chem 51:2227–2231

    Article  CAS  PubMed  Google Scholar 

  • Clarke SF, Guy PL, Burritt DJ, Jameson PE (2002) Changes in the activities of antioxidant enzymes in response to virus infection and hormone treatment. Physiol Plant 114:157–164

    Article  CAS  PubMed  Google Scholar 

  • Durner J, Klessig DF (1997) Inhibition of ascorbato peroxidase by salicylic acid and 2,6-dichloroisonicotinic acid, two inducers of plant defense responses. Proc Natl Acad Sci USA 92:11312–11316

    Article  Google Scholar 

  • El-Shora HM (2002) Properties of phenylalanine ammonia-lyase from marrow cotyledons. Plant Sci 162:1–7

    Article  CAS  Google Scholar 

  • Elvira MI, Galdeano MM, Gilardi P, Garcia-Luque I, Serra MT (2008) Proteomic analysis of pathogenesis-related proteins (PRs) induced by compatible and incompatible interactions of Pepper mild mottle virus (PMMoV) in Capsicum chinense L3 plants. J Exp Bot 59:1253–1265

    Article  CAS  PubMed  Google Scholar 

  • Flexas J, Ribas-Carbó M, Diaz-Espejo A, Galmés J, Medrano H (2008) Mesophyll conductance to CO2: current knowledge and future prospects. Plant Cell Environ 31:602–621

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gay C, Collins J, Gebicki JM (1999) Hydroperoxide assay with the ferric-xylenol orange complex. Anal Biochem 273:149–155

    Article  CAS  PubMed  Google Scholar 

  • Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930

    Article  CAS  PubMed  Google Scholar 

  • Gonçalves MC, Vega J, Oliveira JG, Gomes MMA (2005) Sugarcane yellow leaf virus infection leads to alterations in photosynthetic efficiency and carbohydrate accumulation in sugarcane leaves. Fitopatol Brasil 30:10–16

    Google Scholar 

  • Gonçalves LSA, Rodrigues R, Diz MSS, Robaina RR, Júnior ATA, Carvalho AO, Gomes VM (2013) Peroxidase is involved in Pepper yellow mosaic virus resistance in Capsicum baccatum var. pendulum. Genet Mol Res 12:1411–1420

    Article  PubMed  Google Scholar 

  • Havaux M (1998) Carotenoids as membrane stabilizers in chloroplasts. Trends Plant Sci 3:147–151

    Article  Google Scholar 

  • Havir EA, McHale NA (1987) Biochemical and developmental characterization of multiple forms of catalase in tobacco leaves. Plant Physiol 84:450–455

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hoagland DR, Arnon DI (1950) The Water Culture method for growing plants without soil, vol 347. California Agric. Exp. Station of University of California, Berkeley

    Google Scholar 

  • Hodgson RA, Beachy RN, Pakrasi HB (1989) Selective inhibition of photosystem II in spinach by tobacco mosaic virus: an effect of the viral coat protein. FEBS Lett 245:267–270

    Article  CAS  PubMed  Google Scholar 

  • Iglesias VA, Meins FJ (2000) Movement of plant viruses is delayed in a β-1,3-glucanase-deficient mutagenized showing a reduced plasmodesmatal size exclusion limit and enhanced callose deposition. Plant J 21:157–166

    Article  CAS  PubMed  Google Scholar 

  • Jones JDG, Dangl JL (2006) The plant immune system. Nature 444:323–329

    Article  CAS  PubMed  Google Scholar 

  • Kim YJ, Hwang BK (1994) Differential accumulation of β-1,3-glucanase and chitinase isoforms in pepper stems infected by compatible and incompatible isolates of Phytophthora capsici. Physiol Mol Plant Pathol 45:195–209

    Article  CAS  Google Scholar 

  • Koshiba T (1993) Cytosolic ascorbate peroxidase in seedlings and leaves of maize (Zea mays). Plant Cell Physiol 34:713–721

    CAS  Google Scholar 

  • Krzymowska M, Talarczyk A, Hennig J (1997) Is tobacco response to TMV infection modulated by catalase activity? Acta Physiol Plant 19:577–579

    Article  Google Scholar 

  • Kundu S, Chakraborty D, Kundu A, Pal A (2013) Proteomics approach combined with biochemical attributes to elucidate compatible and incompatible plant-virus interactions between Vigna mungo and Mungbean Yellow Mosaic India Virus. Proteome Sci 11:140–149

    Article  Google Scholar 

  • Laliberté JF, Zheng H (2014) Viral manipulation of plant host membranes. Annu Rev Virol 1:237–259

    Article  PubMed  Google Scholar 

  • Lellis AD, Kasschau KD, Whitham SA, Carrington JC (2002) Loss-of-susceptibility mutagenized of Arabidopsis thaliana reveal an essential role for eIF(iso)4E during potyvirus infection. Curr Biol 12:1046–1051

    Article  CAS  PubMed  Google Scholar 

  • Lichtenthales HK, Wellburn AR (1983) Determinations of total carotenoids and chlorophylls a and b leaf extracts in different solutions. Biochem Soc 11:591–592

    Article  Google Scholar 

  • Liu J, Yang J, Bi H, Zhang P (2014) Why mosaic? Gene expression profiling of African Cassava Mosaic virus-infected cassava reveals the effect of chlorophyll degradation on symptom development. J Integr Plant Biol 56:122–132

    Article  CAS  PubMed  Google Scholar 

  • Loveless A (1958) Increased rate of plaque-type and host-range mutation following treatment of bacteriophage in vitro with ethyl methanesulphonate. Nature (London) 181:1212–1213

    Article  CAS  Google Scholar 

  • Maia JM, Voigt EL, Ferreira-Silva SL, Fontenele AV, Macêdo CEC, Silveira JAG (2013) Differences in cowpea root growth triggered by salinity and dehydration are associated with oxidative modulation involving types I and III peroxidases and apoplastic ascorbate. J Plant Growth Regul 32:376–387

    Article  CAS  Google Scholar 

  • Mlícková K, Luhová L, Lebeda A, Mieslerová B, Pec P (2004) Reactive oxygen species generation and peroxidase activity during Oidium neolycopersici infection on Lycopersicon species. Plant Physiol Biochem 42:753–761

    Article  PubMed  Google Scholar 

  • Molano J, Durán A, Cabib E (1977) A rapid and sensitive assay for chitinase using tritiated chitin. Anal Biochem 83:648–656

    Article  CAS  PubMed  Google Scholar 

  • Mori T, Sakurai M, Sasuta M (2001) Effects of conditioned medium on activities of PAL, CHS, DAHP synthase (DS-Co and Ds-Mn) and anthocyanin production in suspension cultures of Fragaria ananassa. Plant Sci 160:355–360

    Article  CAS  PubMed  Google Scholar 

  • Ngadze E, Icishahayo D, Coutinho TA, van der Waals JE (2012) Role of polyphenol oxidase, peroxidase, phenylalanine ammonia lyase, chlorogenic acid, and total soluble phenols in resistance of potatoes to soft rot. Plant Dis 96:186–192

    Article  CAS  Google Scholar 

  • Pallas V, García JA (2011) How do plant viruses induce disease? Interactions and interference with host components. J Gen Virol 92:2691–2705

    Article  CAS  PubMed  Google Scholar 

  • Passardi F, Penel C, Dunand C (2004) Performing the paradoxical: how plant peroxidases modify the cell wall. Trends Plant Sci 9:534–540

    Article  CAS  PubMed  Google Scholar 

  • Paz CD, Lima JAA, Pio-Ribeiro G, Assis Filho FM, Andrade GP, Gonçalves MFB (1999) Purificação de um isolado do vírus do mosaico severo do caupi, obtido em Pernambuco, produção de antissoros e determinação de fontes de resistência em caupi. Summa Phytopathol 25:285–288

    Google Scholar 

  • Peixoto PHP, Cambraia J, Sant’Anna R, Mosquim PR, Moreira MA (1999) Aluminum effects on lipid peroxidation and on the activities of enzymes of oxidative metabolism in sorghum. Rev Bras Fisiol Veg 11:137–143

    CAS  Google Scholar 

  • Pérez-Bueno ML, Ciscato M, Van de Ven M, García-Luque I, Valcke R, Barón M (2006) Imaging viral infection: studies on Nicotiana benthamiana plants infected with the pepper mild mottle Tobamovirus. Photosynth Res 90:111–123

    Article  PubMed  Google Scholar 

  • Piron F, Nicolaï M, Minoïa S, Piednoir E, Moretti A, Piednoir E, Moretti A, Salgues A, Zamir D, Caranta C, Bendahmane A (2010) An induced mutation in tomato eIF4E leads to immunity to two potyviruses. PLoS One 5:1–10

    Article  Google Scholar 

  • Reissig JL, Storminger JL, Leloir LF (1955) A modified colorimetric method for the estimation of N-acetylamino sugars. J Biol Chem 217:959–966

    CAS  PubMed  Google Scholar 

  • Rys M, Juhasz C, Surowka E, Janeczko A, Saja D, Tobiás I, Skoczowski A, Barna B, Gullner G (2014) Comparison of a compatible and an incompatible pepper-tobamovirus interaction by biochemical and non-invasive techniques: chlorophylla fluorescence, isothermal calorimetry and FT-Raman spectroscopy. Plant Physiol Biochem 83:267–278

    Article  CAS  PubMed  Google Scholar 

  • Scharte J, Schön H, Weis E (2005) Photosynthesis and carbohydrate metabolism in tobacco leaves during an incompatible interaction with Phytophthora nicotianae. Plant, Cell Environ 28:1421–1435

    Article  CAS  Google Scholar 

  • Schreiber U, Bilger W, Neubauer C (1994) Chlorophyll fluorescence as a nonintrusive indicator for rapid assessment of in vivo photosynthesis. In: Schulze ED, Caldwell MM (eds) Ecophysiology of photosynthesis. Springer, Berlin, pp 49–70

    Google Scholar 

  • Shimura H, Pantaleo V, Ishihara T, Myojo N, Inaba J, Sueda K, Burgyán J, Masuta C (2011) A viral satellite RNA induces yellow symptoms on tobacco by targeting a gene involved in chlorophyll biosynthesis using the RNA silencing machinery. PLoS Pathog 7:1–12

    Article  Google Scholar 

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

    Article  Google Scholar 

  • Silveira JAG, Costa RCL, Oliveira JTA (2001) Drought-induced effects and recovery of nitrate assimilation and nodule activity in cowpea plants inoculated with Bradyrhizobium spp. under moderate nitrate level. Braz J Microbiol 32:187–194

    Article  Google Scholar 

  • Soosaar JL, Burch-Smith TM, Dinesh-Kumar SP (2005) Mechanisms of plant resistance to viruses. Nat Rev Microbiol 3:789–798

    Article  CAS  PubMed  Google Scholar 

  • Torres MA (2010) ROS in biotic interactions. Physiol Plant 138:414–429

    Article  CAS  PubMed  Google Scholar 

  • Urbanek H, Kuzniak-Gebarowska E, Herka K (1991) Elicitation of defense responses in bean leaves by Botrytis cinerea polygalacturonase. Acta Physiol Plant 13:43–50

    CAS  Google Scholar 

  • Van Loon LC, Rep M, Pieterse CM (2006) Significance of inducible defense-related proteins in infected plants. Annu Rev Phytopathol 44:135–162

    Article  PubMed  Google Scholar 

  • Van Rossum MWPC, Alberda M, Van der Plas LHW (1997) Role of oxidative damage in tulip bulb scale micropropagation. Plant Sci 130:207–216

    Article  Google Scholar 

  • Vanholme R, Demedts B, Morreel K, Ralph J, Boerjan W (2010) Lignin biosynthesis and structure. Plant Physiol 153:895–905

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

Council for Advanced Professional Training (CAPES) sponsored Pedro F. N. Souza with a doctoral grant. National Council for Scientific and Technological Development (CNPq, grant 308107/2013-6), Research Council of the State of Ceara (FUNCAP, grant 2155/PRONEX), and Cearense Federation of Research and Culture (FCPC, Grant 09/2008) financially supported this work.

Author information

Authors and Affiliations

  1. Laboratory of Plant Defense Proteins, Department of Biochemistry and Molecular Biology, Federal University of Ceara, Av. Mister Hull, P.O. Box: 60451, Fortaleza, CE, Brazil

    Pedro F. N. Souza, Fredy D. A. Silva, Fabricio E. L. Carvalho, Joaquim A. G. Silveira, Ilka M. Vasconcelos & Jose T. A. Oliveira

Authors
  1. Pedro F. N. Souza
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fredy D. A. Silva
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Fabricio E. L. Carvalho
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Joaquim A. G. Silveira
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ilka M. Vasconcelos
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jose T. A. Oliveira
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jose T. A. Oliveira.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by C.-H. Dong.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (TIFF 1893 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Souza, P.F.N., Silva, F.D.A., Carvalho, F.E.L. et al. Photosynthetic and biochemical mechanisms of an EMS-mutagenized cowpea associated with its resistance to cowpea severe mosaic virus . Plant Cell Rep 36, 219–234 (2017). https://doi.org/10.1007/s00299-016-2074-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-016-2074-z

Keywords

Search

Navigation