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In planta validation of nitric oxide mediated defense responses in common bean against Colletotrichum gloeosporioides infection

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Abstract

Our present study demonstrates an in planta approach of defense responses in bean plants against the devastating pathogen Colletotrichum gloeosporioides (Penz.) Penz. and Sacc. Recent evidences suggest the function of nitric oxide (NO) as a strong signaling molecule in many plant pathogen interactions. Standing at this point, the effect of NO durability in activation plant defense is poorly understood. Progression and establishment of the pathogen in the host has been confirmed by bright field and scanning electron microscopy. Biochemical studies indicated that during the pathogenesis, the NO content induced up to 12 days after pathogen inoculation, then gradually declined. Induction in NO content was positively correlated with different defense related enzyme activities and antimicrobial compounds like, total phenol and total flavonoid content. Chlorophyll content was also reduced as the pathogen progressed gradually. Lipid peroxidation and proline content are important stress-related markers were thus also estimated. Cell death was also found to increase with disease progression. Overall study suggests that during early stages of pathogen progression NO produce rapidly to intensify host defense but when virulent pathogen well established in the host cell NO production hampers significantly and as a consequence host defense mechanism also become down-regulated. So, durability and maintenance of NO level is one of the key factors of host resistance against pathogen.

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References

  • Acharya K, Chakraborty N, Dutta AK, Sarkar S, Acharya R (2011) Signaling role of nitric oxide in the induction of plant defense by exogenous application of abiotic inducers. Arch Phytopathol Plant Protect 44:1501–1511

    Article  CAS  Google Scholar 

  • Agrios GN (2005) Plant pathology, 5th edn. Academic Press, New York

    Google Scholar 

  • Ahmad L, Siddiqui ZA, Abd-Allah EF (2019) Effects of interaction of Meloidogyne incognita, Alternaria dauci and Rhizoctonia solani on the growth, chlorophyll, carotenoid and proline contents of carrot in three types of soil. Acta Agric Scand Sect B Soil Plant Sci 69:324–331

    CAS  Google Scholar 

  • Anaruma ND, Schmidt FL, Duarte MCT, Figueira GM, Delarmelina C, Benato EA, Sartoratto A (2010) Control of Colletotrichum gloeosporioides (penz.) Sacc. in yellow passion fruit using Cymbopogon citrates essential oil. Braz J Microbiol 41:66–73

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–15

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ayala A, Muñoz MF, Argüelles S (2014) Lipid peroxidation: production, metabolism and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxid Med Cell Lingev 2014:360438

    Google Scholar 

  • Barampama Z, Siard RE (1993) Nutrient composition, protein quality and antinutritional factors of some varieties of dry beans (Phaseolus vulgaris) grown in Burundi. Food Chem 47:159–167

    Article  CAS  Google Scholar 

  • Bates LS, Waldran RP, Teare ID (1973) Raipid determination of free proline for water studies. Plant Soil 39:205–208

    Article  CAS  Google Scholar 

  • Beligni MV, Lamattina L (2001) Nitric oxide in plants: the history is just beginning. Plant Cell Environ 24:267–278

    Article  CAS  Google Scholar 

  • Bethke PC, Libourel IGL, Jones RL (2006) Nitric oxide reduces seed dormancy in Arabidopsis. J Exp Bot 57:517–526

    Article  CAS  PubMed  Google Scholar 

  • Bethke PC, Libourel IGL, Aoyama N, Chung Y, Still DW, Jones RL (2007) The Arabiodopsis thaliana aleurone layer responds to nitric oxide, gibberellins and abscisic acid and is sufficient and necessary for seed dormancy. Plant Physiol 143:1173–1188

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Cakmak I, Horst J (1991) Effect of aluminium on lipid peroxidation, superoxide dismutase, catalase, and peroxidase activities in root tips of soybean (Glycine max). Physiol Planta 83:463–468

    Article  CAS  Google Scholar 

  • Canno PF, Damm U, Johnston PR, Weir BS (2012) Colletotrichum—current status and future directions. Stud Mycol 73:181–213

    Article  Google Scholar 

  • Celmeli T, Sari H, Canci H, Sari D, Adak A, EkerT, Toker C (2018) The nutritional content of common bean (Phaseolus vulgaris L.) landraces in comparison to modern varieties. Agronomy 8:166

    Article  CAS  Google Scholar 

  • Chakraborty N, Acharya K (2017) “NO way”! Says the plant to abiotic stress. Plant Gene 11:99–105

    Article  CAS  Google Scholar 

  • Chakraborty N, Chandra S, Acharya K (2015) Sublethal heavy metal stress stimulates innate immunity in tomato. Sci World J 2015:208649

    Article  Google Scholar 

  • Chakraborty N, Ghosh S, Chandra S, Sengupta S, Acharya K (2016) Abiotic elicitors mediated elicitation of innate immunity in tomato: an ex vivo comparison. Physiol Mol Biol Plants 22:307–320

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chakraborty N, Chandra S, Acharya K (2017) Biochemical basis of improvement of defense in tomato plant against Fusarium wilt by CaCl2. Physiol Mol Biol Plants 23:581–596

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chakraborty N, Mukherjee K, Sarkar A, Acharya K (2019) Interaction between bean and Colletotrichum gloeosporioides: understanding through a biochemical approach. Plants 8:345

    Article  CAS  PubMed Central  Google Scholar 

  • Chakraborty N, Banerjee M, Acharya K (2020) In vitro selection of elite clone of Withania somnifera against leaf blight disease caused by Alternaria alternata. Physiol Mol Plant Pathol 112:101560

    Article  CAS  Google Scholar 

  • Chandra S, Chakraborty N, Chakraborty A, Rai R, Bera B, Acharya K (2014) Abiotic elicitor-mediated improvement of innate immunity in Camellia sinensis. J Plant Growth Regul 33:849–859

    Article  CAS  Google Scholar 

  • Chandra S, Chakraborty N, Dasgupta A, Sarkar J, Panda K, Acharya K (2015) Chitosan nanoparticles: a positive modulator of innate immune responses in plants. Sci Rep 5:15195

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chang C-C, Yang M-H, Wen H-M, Chern J-C (2002) Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 10:178–182

    CAS  Google Scholar 

  • Cheng GW, Breen PJ (1991) Activity of phenylalanine ammonia-lyase (PAL) and concentrations of anthocyanins and phenolics in developing strawberry fruit. J Am Soc Hortic Sci 116:865–869

    Article  CAS  Google Scholar 

  • Collinge DB, Gregersen PL, Thordal-Christensen H (1994) The induction of gene expression in response to pathogenic microbes. In: Basra AS (ed) Mechanisms of plant growth and productivity: modern approaches and perspectives. Marcel Dekker, New York, pp 391–433

    Google Scholar 

  • Delledonne M, Zeier J, Marocco A, Lamb C (2001) Signal interactions between nitric oxide and reactive oxygen intermediates in the plant hypersensitive disease resistance response. Proc Natl Acad Sci 98:13454–13459

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Deng L, Zeng K, Zhou Y, Huang Y (2015) Effects of postharvest oligochitosan treatment on anthracnose disease in citrus (Citrus sinensis L. Osbeak) fruit. Eur Food Res Technol 240:795–804

    Article  CAS  Google Scholar 

  • Dickerson DP, Pascholati SF, Hagerman AE, Butler LG, Nicholson RL (1984) Phenylalanine ammonia-lyase and hydroxycinnamate: CoA ligase in maize mesocotyls inoculated with Helminthosporium maydis or Helminthosporium carbonum. Physiol Plant Pathol 25:111–123

    Article  CAS  Google Scholar 

  • Edirisinghe M, Ali A, Maqbool M, Alderson PG (2014) Chitosan controls postharvest anthracnose in bell pepper by activating defense-related enzymes. J Food Sci Technol 51:4078–4083

    Article  CAS  PubMed  Google Scholar 

  • Ge Y, Bi Y, Guest DI (2013) Defence responses in leaves of resistant and susceptible melon (Cucumis melo L.) cultivars infected with Colletotrichum lagenarium. Physiol Mol Plant Pathol 81:13–21

    Article  CAS  Google Scholar 

  • Gowtham HG, Murali M, Brijesh Singh S, Lakshmeesha TR, Narasimha Murthy K, Amruthesh KN, Niranjana SR (2018) Plant growth promoting rhizobacteria—Bacillus amyloliquefaciens improves plant growth and induces resistance in chilli against anthracnose disease. Biologic cont 126:209–217

    CAS  Google Scholar 

  • Gutteridge JM (1995) Lipid peroxidation and antioxidants as biomarkers of tissue damage. Clin chem 41:1819–1828

    Article  CAS  PubMed  Google Scholar 

  • Guzmán-Maldonado SH, Acosta-Gallegos J, Paredes‐López O (2000) Protein and mineral content of a novel collection of wild and weedy common bean (Phaseolus vulgaris L). J Sci Food Agric 80:1874–1881

    Article  Google Scholar 

  • Hayat S, Hayat Q, Alyemeni MN, Wani AS, Pichtel J, Ahmad A (2012) Role of proline under changing environments. Plant Signal Behav 7:1456–1466

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hemeda HM, Klein BP (1990) Effects of naturally occurring antioxidants on peroxidase activity of vegetable extracts. J Food Sci 55:184–185

    Article  CAS  Google Scholar 

  • Koch E, Slusarenko A (1990) Arabidopsis is susceptible to infection by a downy mildew fungus. Plant Cell 2:437–445

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kumar K, Khan P (1982) Peroxidase and polyphenol oxidase in excised ragi (Eleusine corocana cv PR 202) leaves during senescence. Indian J Exp Biol 20:412–416

    CAS  Google Scholar 

  • Kumari PR, Singh R (2017) Anthracnose of mango incited by Colletotrichum gloeosporioides: a comprehensive review. Int J Pure App Biosci 5:48–56

    Article  Google Scholar 

  • Libourel IGL, Van Bodegom PM, Fricker MD, Ratcliffe RG (2006) Nitrite reduces cytoplasmic acidosis under anoxia. Plant Physiol 142:1710–1717

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu R, Xu S, Li J, Hu Y, Lin Z (2006) Expression profile of PAL gene from Astragalus membranaceus var. mongholicus and its crucial role in flux into flavonoid biosynthesis. Plant Cell Rep 25:705–710

    Article  CAS  PubMed  Google Scholar 

  • Manzo D, Ferriello F, Puopolo G, Zoina A, D’Esposito D, Tardella L, Ferrarini A, Ercolano MR (2016) Fusarium oxysporum f. sp. radicis-lycopersici induces distinct transcriptome reprogramming in resistant and susceptible isogenic tomato lines. BMC Plant Biol 16:53

    Article  PubMed  PubMed Central  Google Scholar 

  • Marletta MA, Spiering MM (2003) Trace elements and nitric oxide function. J Nutr 133:1431–1433

    Article  Google Scholar 

  • Mohanty SK, Sridhar R (1982) Physiology of rice tungro virus disease: proline accumulation due to infection. Physiol Planta 56:89–93

    Article  CAS  Google Scholar 

  • Mylonas C, Kouretas D (1999) Lipid peroxidation and tissue damage. Vivo 13:295–309

    CAS  Google Scholar 

  • Nisha S, Revathi K, Chandrasekaran R, Kirubakaran SA, Sathish-Narayanan S, Stout MJ, Senthil-Nathan S (2012) Effect of plant compounds on induced activities of defense-related enzymes and pathogenesis related protein in bacterial blight disease susceptible rice plant. Physiol Mol Plant Pathol 80:1–9

    Article  CAS  Google Scholar 

  • Pan SQ, Ye XS, Kuć J (1991) Association of β-1,3-glucanase activity and isoform pattern with systemic resistance to blue mould in tobacco induced by stem injection with Peronospora tabacina or leaf inoculation with tobacco mosaic virus. Physiol Mol Plant Pathol 39:25–39

    Article  CAS  Google Scholar 

  • Piršelová B, Matušiková I (2013) Callose: the plant cell wall polysaccharide with multiple biological functions. Acta Physiol Plant 35:635–644

    Article  Google Scholar 

  • Rajapakse RGAS, Ranasinghe JADAR (2002) Development of variety screening method for anthracnose disease of chilli (Capsicum annum L.) under field conditions. Trop Agric Res Ext 5:7–11

    Google Scholar 

  • Ranathunge NP, Mongkolporn O, Ford R, Taylor PWJ (2012) Colletotrichum truncatum pathosystem on Capsicum spp: infection, colonization and defence mechanisms. Austral Plant Pathol 41:463–473

    Article  Google Scholar 

  • Romero-Puertas MC, Perazzolli M, Zago ED, Delledonne M (2004) Nitric oxide signalling functions in plant-pathogen interactions. Cell Microbiol 6:795–803

    Article  CAS  PubMed  Google Scholar 

  • Sarkar A, Chakraborty N, Acharya K (2021) Unraveling the role of nitric oxide in regulation of defense responses in chilli against Alternaria leaf spot disease. Physiol Mol Plant Pathol 114:101621

    Article  CAS  Google Scholar 

  • Schenk ST, Schikora A (2015) Staining of callose depositions in root and leaf tissues. Bio-Protocol 5:e1429

    Article  Google Scholar 

  • Siddiqui ZA, Parveen A, Ahmad L, Hashem A (2019) Effects of graphene oxide and zinc oxide nanoparticles on growth, chlorophyll, carotenoids, proline contents and diseases of carrot. Sci Hortic 249:374–382

    Article  CAS  Google Scholar 

  • Silva KJD, Souza EA, Ishikawa FH (2007) Characterization of Colletotrichum lindemuthianum isolates from the state of Minas Gerais, Brazil. J Phytopathol 155:241–247

    Article  Google Scholar 

  • Tu JC (1983) Epidemiology of anthracnose caused by Colletotrichum lindemuthiaum on white bean (Phaseolus vulgaris) in southern Ontario: survival of the pathogen. Plant Dis 67:402–404

    Article  Google Scholar 

  • Van Loon LC, Bakker PAHM, Pieters CMJ (1998) Systemic resistance induced by rhizosphere bacteria. Annu Rev Phytopathol 36:453–483

    Article  PubMed  Google Scholar 

  • Waller JM, Lenné JM, Waller SJ (2002) Plant pathologist’s pocketbook, 3rd edn. CABI, Wallingford

    Google Scholar 

  • Wang Y, Loake GJ, Chu C (2013) Cross-talk of nitric oxide and reactive oxygen species in plant programed cell death. Front Plant Sci 4:314

    Article  PubMed  PubMed Central  Google Scholar 

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

    CAS  Google Scholar 

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Acknowledgements

The authors declare their inexplicable gratitude toward Dr. Shampa Bhattacharyya and Dr. Rajyasri Ghosh, Post Graduate Department of Botany, Scottish Church College, Kolkata, for providing us fungal and plant samples with which this study has been performed.

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This research did not receive any external funding.

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Authors and Affiliations

  1. Department of Botany, Scottish Church College, Kolkata, 700006, India

    Nilanjan Chakraborty, Anamika Paul & Kabita Mukherjee

  2. Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, Kolkata, 700019, India

    Anik Sarkar, Adhiraj Dasgupta & Krishnendu Acharya

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  1. Nilanjan Chakraborty
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  2. Anik Sarkar
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  3. Adhiraj Dasgupta
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  4. Anamika Paul
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  6. Krishnendu Acharya
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Conceptualization, NC; methodology and investigation, AS and KM; writing—original draft preparation, AP and NC; writing—review and editing, KA and ADG.

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Correspondence to Nilanjan Chakraborty.

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Chakraborty, N., Sarkar, A., Dasgupta, A. et al. In planta validation of nitric oxide mediated defense responses in common bean against Colletotrichum gloeosporioides infection. Indian Phytopathology 75, 15–24 (2022). https://doi.org/10.1007/s42360-021-00425-0

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