Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Synergistic effects of silver nanoparticles augmented Calothrix elenkinii for enhanced biocontrol efficacy against Alternaria blight challenged tomato plants

  • 14 Accesses


The biocontrol efficacy of a cyanobacterium Calothrix elenkinii (Ce), silver nanoparticles (AgNPs) and their augmented complex (AgNPs-Ce) was evaluated. Foliar application of AgNPs-Ce reduced the disease severity by 47–58%, along with significant increases of 44–45%, 40–46% and 23–33% in leaf chlorophyll, carotenoid content, and polyphenol oxidase activity in the A. alternata infected tomato plants. A significant reduction in the pathogen load was recorded, both by plate counts and microscopic observations in the AgNPs, Ce and AgNPs-Ce treatments, while AgNPs-Ce also effectively reduced ergosterol content by 63–79%. Amplification using PCR-ITS primers revealed very faint bands or none in the AgNPs-Ce treated leaves, illustrating the inhibition of fungal growth. Significantly higher yield was recorded in the pathogen challenged plants receiving AgNPs-Ce, AgNPs, and Ce treatments. Higher expression of elicited antioxidant enzymes, along with enhanced plant growth attributes and lowered fungal load highlight the biocontrol potential of AgNPs-Ce treatment in A. alternata infected plants. This synergistic association can be explored as a promising biocontrol option against A. alternata challenged tomato plants under various agroclimatic conditions.

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

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


  1. Abdel-Hafez SI, Nafady NA, Abdel-Rahim IR et al (2016) Assessment of protein silver nanoparticles toxicity against pathogenic Alternaria solani. Biotech 6:199

  2. Adhikari P, Oh Y, Panthee DR (2017) Current status of early blight resistance in tomato: an update. Int J Mol Sci 18:2019

  3. Babu S, Bidyarani N, Chopra P et al (2015) Evaluating microbe-plant interactions and varietal differences for enhancing biocontrol efficacy in root rot disease challenged cotton crop. Eur J Plant Pathol 142:345–362

  4. Baiyee B, Ito SI, Sunpapao A (2019) Trichoderma asperellum T1 mediated antifungal activity and induced defense response against leaf spot fungi in lettuce (Lactuca sativa L.). Physiol Mol Plant Pathol 106:96–101

  5. Berger S, Papadopoulos M, Schreiber U et al (2004) Complex regulation of gene expression, photosynthesis and sugar levels by pathogen infection in tomato. Physiol Plant 122:419–428

  6. Chaudhary V, Prasanna R, Nain L et al (2012) Bioefficacy of novel cyanobacteria-amended formulations in suppressing damping off disease in tomato seedlings. World J Biotechnol 28:3301–3310

  7. Chung KR (2014) Reactive oxygen species in the citrus fungal pathogen Alternaria alternata: the roles of NADPH-dependent oxidase. Physiol Mol Plant Pathol 88:10–17

  8. Dawson-Andoh BE (2002) Ergosterol content as a measure of biomass of potential biological control fungi in liquid cultures. Holz als Roh-und Werkst 60:115–117

  9. Dixit R, Agrawal L, Gupta S et al (2016) Southern blight disease of tomato control by 1-aminocyclopropane-1-carboxylate (ACC) deaminase producing Paenibacillus lentimorbus B-30488. Plant Signal Behav 11:e1113363

  10. Elmer WH, Ma C, White JC (2018) Nanoparticles for plant disease management. Curr Opin Environ Sci Health 6:66

  11. Fang C, Zhuang Y, Xu T et al (2013) Changes in rice allelopathy and rhizosphere microflora by inhibiting rice phenylalanine ammonia-lyase gene expression. J Chem Ecol 39:204–212

  12. Gold K, Slay B, Knackstedt M, Gaharwar AK (2018) Antimicrobial activity of metal and metal-oxide based nanoparticles. Adv Ther 1:1700033

  13. Gorczyca A, Pociecha E, Kasprowicz M, Niemiec M (2015) Effect of nanosilver in wheat seedlings and Fusarium culmorum culture systems. Eur J Plant Pathol 142:251–261

  14. Hagmann L, Jiittner F (1996) Fischerellin A, a novel photosystem-ll-inhibiting allelochemical of the cyanobacterium Fischerella muscicola with antifungal and herbicidal activity. Tetrahedron Lett 37:6539–6542

  15. Hata M, Ishii Y, Watanabe ERI et al (2010) Inhibition of ergosterol synthesis by novel antifungal compounds targeting C-14 reductase. Med Mycol 48:613–621

  16. Ighodaro OM, Akinloye OA (2018) First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): their fundamental role in the entire antioxidant defence grid. Alex Med J 54:287–293

  17. Jennings PH, Brannaman BL, Zscheile FP (2019) Peroxidase and polyphenol oxidase activity associated with Helminthosporium leaf spot of maize. Phytopathology 59:963–967

  18. Khan N, Mishra A, Nautiyal CS (2012) Paenibacillus lentimorbus B-30488r controls early blight disease in tomato by inducing host resistance associated gene expression and inhibiting Alternaria solani. Biol Control 62:65–74

  19. Kreitlow S, Mundt S, Lindequist U (1999) Cyanobacteria—a potential source of new biologically active substances. J Biotechnol 70:61–63

  20. Krome K, Kabsch U, Aumann J (2016) The effect of benzothiadiazole and fungal extracts of Cercospora beticola and Fusarium graminearum on phosphoenolpyruvate carboxylase activity in cucumber leaves. J Plant Dis Prot 114:250–255

  21. Kumari M, Pandey S, Bhattacharya A et al (2017) Protective role of biosynthesized silver nanoparticles against early blight disease in Solanum lycopersicum. Plant Physiol Biochem 121:216–225

  22. Kuvalekar A, Redkar A, Gandhe K, Harsulkar A (2011) Peroxidase and polyphenol oxidase activities in compatible host-pathogen interaction in Jasminum officinale and Uromyces hobsoni: insights into susceptibility of host. N Z J Bot 49:351–359

  23. Lamsal K, Kim SW, Jung JH et al (2011) Application of silver nanoparticles for the control of Colletotrichum species in vitro and pepper anthracnose disease in field. Mycobiology 39:194–199

  24. Latifi A, Ruiz M, Zhang C (2009) Oxidative stress in cyanobacteria. FEMS Microbiol Rev 33:258–278

  25. Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382

  26. Lobato AKS, Gonçalves-Vidigal MC, Vidigal Filho PS et al (2010) Relationships between leaf pigments and photosynthesis in common bean plants infected by anthracnose. N Z J Crop Hortic Sci 38:29–37

  27. Magyarosy AC, Schurmann P, Buchanan BB (1975) Effect of powdery mildew infection on photosynthesis by leaves and chloroplasts of sugar beets. Plant Physiol 57:486–489

  28. Mahawar H, Prasanna R (2018) Prospecting the interactions of nanoparticles with beneficial microorganisms for developing green technologies for agriculture. Environ Nanotechnol Monit Manag 10:477–485

  29. Mahawar H, Prasanna R, Singh SB, Nain L (2018) Influence of silver, zinc oxide and copper oxide nanoparticles on the cyanobacterium Calothrix elenkinii. BioNanoScience 8:802–810

  30. Mai VC, Nguyen BH, Nguyen DD (2017) Nostoc calcicola extract improved the antioxidative response of soybean to cowpea aphid. Bot Stud 58:55

  31. Manjunath M, Prasanna R, Nain L et al (2010) Biocontrol potential of cyanobacterial metabolites against damping off disease caused by Pythium aphanidermatum in solanaceous vegetables. Arch Phytopathol Plant Prot 43:666–667

  32. Manjunath M, Kanchan A, Ranjan K et al (2016) Beneficial cyanobacteria and eubacteria synergistically enhance bioavailability of soil nutrients and yield of okra. Heliyon 2:e00066

  33. Marslin G, Sheeba CJ, Franklin G (2017) Nanoparticles alter secondary metabolism in plants via ROS burst. Front Plant Sci 8:1–8

  34. Miranda M, Ralph SG, Mellway R et al (2007) The transcriptional response of hybrid poplar (Populus trichocarpa × P. deltoides) to infection by melampsora medusae leaf rust involves induction of flavonoid pathway genes leading to the accumulation of proanthocyanidins. Mol Plant Microbe Interact 20:816–831

  35. Natarajan C, Prasanna R, Gupta V et al (2012) Characterization of the fungicidal activity of Calothrix elenkinii using chemical methods and microscopy. Appl Biochem Microbiol 48:59–65

  36. Ng HE, Raj SSA, Wong SH et al (2008) Estimation of fungal growth using the ergosterol assay: a rapid tool in assessing the microbiological status of grains and feeds. Lett Appl Microbiol 46:113–118

  37. O’Leary B, Park J, Plaxton WC (2011) The remarkable diversity of plant PEPC (phosphoenolpyruvate carboxylase): recent insights into the physiological functions and post-translational controls of non-photosynthetic PEPCs. Biochem J 436:15–34

  38. Ouda SM (2014) Antifungal activity of silver and copper nanoparticles on two plant pathogens, Alternaria alternata and Botrytis cinerea. Res J Microbiol 9:34–42

  39. Pane C, Zaccardelli M (2015) Evaluation of Bacillus strains isolated from solanaceous phylloplane for biocontrol of Alternaria early blight of tomato. Biol Control 84:11–18

  40. Prabhu S, Poulose EK (2012) Silver nanoparticles: mechanism of antimicrobial action, synthesis, medical applications, and toxicity effects. Int Nano Lett 2:1–10

  41. Prasanna R, Nain L, Tripathi R et al (2008) Evaluation of fungicidal activity of extracellular filtrates of cyanobacteria—possible role of hydrolytic enzymes. J Basic Microbiol 48:186–194

  42. Prasanna R, Chaudhary V, Gupta V et al (2013) Cyanobacteria mediated plant growth promotion and bioprotection against Fusarium wilt in tomato. Eur J Plant Pathol 136:337–353

  43. Prasanna R, Babu S, Bidyarani N et al (2015) Prospecting cyanobacteria-fortified composts as plant growth promoting and biocontrol agents in cotton. Exp Agric 51:42–65

  44. Rosati RG, Lario LD, Hourcade ME et al (2018) Plant science primary metabolism changes triggered in soybean leaves by Fusarium tucumaniae infection. Plant Sci 274:91–100

  45. Ruiz-Romero P, Valdez-Salas B, González-Mendoza D, Mendez-Trujillo V (2018) Antifungal effects of silver phytonanoparticles from Yucca shilerifera against strawberry soil-borne pathogens: Fusarium solani and Macrophomina phaseolina. Mycobiology 46:47–51

  46. Shilpi G, Shilpi S, Sunita S (2015) Tolerance against heavy metal toxicity in cyanobacteria: role of antioxidant defense system. Int J Pharm Pharm Sci 7:9–16

  47. Simranjit K, Kanchan A, Prasanna R et al (2019) Microbial inoculants as plant growth stimulating and soil nutrient availability enhancing options for cucumber under protected cultivation. World J Microbiol Biotechnol 35:51

  48. Singh PK, Rai S, Pandey S et al (2012) Cadmium and UV-B induced changes in proteomic and some biochemical attributes of Anabaena sp. PCC7120. Phykos 42:39–50

  49. Singh A, Sarma BK, Upadhyay RS, Singh HB (2013) Compatible rhizosphere microbes mediated alleviation of biotic stress in chickpea through enhanced antioxidant and phenylpropanoid activities. Microbiol Res 168:33–40

  50. Stanier RY, Kunisawa R, Mandel M, Cohen-Bazire G (1971) Purification and properties of unicellular blue-green algae (order Chroococcales). Bacteriol Rev 35:171–205

  51. Strzalka K, Kostecka-guga A, Latowski D (2003) Carotenoids and environmental stress in plants: significance of carotenoid-mediated modulation of membrane physical properties. Russ J Plant Physiol 50:168–172

  52. Tarafdar JC, Raliya R (2013) ZnO nanoparticle biosynthesis and its effect on phosphorous-mobilizing enzyme secretion and gum contents in clusterbean (Cyamopsis tetragonoloba L.). Agric Res 2:48–57

  53. Torres MA, Jones JDG, Dangl JL (2006) Reactive oxygen species signaling in response to pathogens. Plant Physiol 141:373–378

  54. Tripathi DK, Tripathi A, Singh S, Singh Y (2017) Uptake, accumulation and toxicity of silver nanoparticle in autotrophic plants, and heterotrophic microbes: a concentric review. Front Microbiol 8:1–16

  55. Waygood ER, Pao LY, Godavari HR (1974) Stimulation of phosphoenolpyruvate carboxylase activity in rust-infected wheat leaves rust. Experientia 30:986–988

  56. Worrall EA, Hamid A, Mody KT et al (2018) Nanotechnology for plant disease management. Agronomy 8:1–24

  57. Wu M-X, Wedding RT (1985) Diurnal regulation of phosphoenolpyruvate carboxylase from Crassula. Plant Physiol 77:667–675

  58. Yadav OP, Dabbas MR, Gaur LB (2014) Screening of tomato advanced lines, genotypes against Alternaria solani. Plant Arch 14:553–555

  59. Yan A, Chen Z (2019) Impacts of silver nanoparticles on plants: a focus on the phytotoxicity and underlying mechanism. Int J Mol Sci 20:1003

Download references


The study was supported by the UGC-DAE Consortium for Scientific Research, University Grants Commission who provided fellowship and the Post Graduate School, ICAR-IARI, who provided the facilities towards the fulfilment of Ph.D. program. The study was also partly funded by the AMAAS Network Project on Microorganisms, granted by the Indian Council of Agricultural Research (ICAR), New Delhi, to RP. We thank the Central Research Facility, IIT, Delhi for helping with the ICP analyses of our samples. We are thankful to the Division of Microbiology, National Phytotron Facility, Division of Plant Pathology, Division of Agricultural Chemicals and Division of Nematology, ICAR-IARI, New Delhi, for providing necessary facilities for undertaking this study.

Author information

All the authors have gone through the final manuscript and agree to its contents and its publication.

Correspondence to Radha Prasanna.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Mahawar, H., Prasanna, R., Gogoi, R. et al. Synergistic effects of silver nanoparticles augmented Calothrix elenkinii for enhanced biocontrol efficacy against Alternaria blight challenged tomato plants. 3 Biotech 10, 102 (2020).

Download citation


  • Alternaria blight
  • Biocontrol
  • Cyanobacterium
  • Nanoparticles
  • Tomato