Abstract
Two novel and rapid in vitro agar plate methods for simultaneous screening of two bacterial isolates for antagonism against multiple fungal phytopathogens in a single agar plate are described. In the modified plus streak method (MPS), in a 10 cm diameter PDA agar plate, two different bacterial isolates were screened against four fungal pathogens. The second method used 20 cm diameter PDA agar plate surface and was termed as the parallel multiple streak (PMS) method. Here, two bacterial isolates were screened for antagonism against ten fungal phytopathogens on a single agar plate surface. Our new methods are rapid and more economical, and could reduce the number of agar plates and the medium required for in vitro screening by several fold, compared to that used in the conventional dual culture plate assay.
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References
Ali S, Hameed S, Shahid M, Iqbal M, Lazarovits G, Imran A (2020) Functional characterization of potential PGPR exhibiting broad-spectrum antifungal activity. Microbiol Res 232:126389. https://doi.org/10.1016/j.micres.2019.126389
Anith KN, Radhakrishnan NV, Manomohandas TP (2002) Management of nursery wilt of black pepper (Piper nigrum L.) with antagonistic bacteria. Curr Sci 83:561–562
Anith KN, Radhakrishnan NV, Manomohandas TP (2003) Screening of antagonistic bacteria for biological control of nursery wilt of black pepper (Piper nigrum). Microbiol Res 158:91–97
Anith KN, Nysanth NS, Natarajan C (2021) Novel and rapid agar plate methods for in vitro assessment of bacterial biocontrol isolates’ antagonism against multiple fungal phytopathogens. Lett Appl Microbiol 73:229–236
Athira S, Anith KN (2020) Plant growth promotion and suppression of bacterial wilt incidence in tomato by rhizobacteria, bacterial endophytes and the root endophytic fungus Piriformospora indica. Indian Phytopathol 73:629–642
Beneduzi A, Ambrosini A, Passaglia LMP (2012) Plant growth-promoting rhizobacteria (PGPR): their potential as antagonists and biocontrol agents. Genet Mol Biol 35:1044–1051
Daayf F, Adam L, Fernando WGD (2003) Comparative screening of bacteria for biological control of potato late blight (strain US-8), using in vitro, detached-leaves, and whole-plant testing systems. Can J Plant Pathol 25:276–284
Dinu A, Kumar A, Aravind R, Eapen SJ (2007) An improved method for selection of antagonistic bacteria against Phytophthora capsici Leonian infections in black pepper (Piper nigrum L.). J Spices Aromat Crops 16:1–7
Fridlender M, Inbar J, Chet I (1993) Biological control of soilborne plant pathogens by a β-1,3 glucanase-producing Pseudomonas cepacia. Soil Biol Biochem 25:1211–1221
Hass D, Defago G (2005) Biological control of soil-borne pathogens by fluorescent pseudomonads. Nat Rev Microbiol 3:307–319
Hassan N, Nakasuji S, Elsharkawy MM, Naznin HA, Kubota M, Ketta H, Shimizu M (2017) Biocontrol potential of an endophytic Streptomyces sp. strain MBCN152-1 against Alternaria brassicicola on cabbage plug seedlings. Microbes Environ 32:133–141
Kloepper JW, Schroth MN (1981) Relationship of in vitro antibiosis of plant growth-promoting rhizobacteria to plant growth and the displacement of root microflora. Phytopathol 71:1020–1024
Kollakkodan N, Anith KN, Radhakrishnan NV (2017) Diversity of endophytic bacteria from Piper spp. with antagonistic property against Phytophthora capsici causing foot rot disease in black pepper (Piper nigrum L.). J Trop Agri 55:63–70
Kollakkodan N, Anith KN, Nysanth NS (2021) Endophytic bacteria from Piper colubrinum suppress Phytophthora capsici infection in black pepper (Piper nigrum L.) and improve plant growth in the nursery. Arch Phytopathol Plant Prot 54:86–108
Larkin RP, Fravel DR (1998) Efficacy of various fungal and bacterial biocontrol organisms for control of Fusarium wilt of tomato. Plant Dis 82:1022–1028
O’Brien PA (2017) Biological control of plant diseases. Australas Plant Pathol 46:293–304
Paul T, Nysanth NS, Yashaswini MS, Anith KN (2021) Inoculation with bacterial endophytes and the fungal root endophyte, Piriformospora indica improves plant growth and reduces foliar infection by Phytophthora capsici in black pepper. J Trop Agric 59:224–235
Pliego C, Ramos C, de Vicente A, Cazorla FM (2011) Screening for candidate bacterial biocontrol agents against soilborne fungal plant pathogens. Plant Soil 340:505–520
Raaijmakers JM, de Bruijn I, Nybroe O, Ongena M (2010) Natural functions of lipopeptides from Bacillus and Pseudomonas: more than surfactants and antibiotics. FEMS Microbiol Rev 34:1037–1062
Raymaekers K, Ponet L, Holtappels D, Berckmans B, Cammue BP (2020) Screening for novel biocontrol agents applicable in plant disease management—a review. Biol Control. https://doi.org/10.1016/j.biocontrol.2020.104240
Santoyo G, Sánchez-Yáñez JM, de los Santos-Villalobos S (2019) Methods for detecting biocontrol and plant growth-promoting traits in Rhizobacteria. In: Methods in rhizosphere biology research, Springer, Singapore, p 133–149. https://doi.org/10.1007/978-981-13-5767-1_8
Schroth MN, Hancock JG (1982) Disease suppressive soil and root colonizing bacteria. Science 216:1376–1381
Tanya A, Filion M (2017) Biocontrol through antibiosis: exploring the role played by sub inhibitory concentrations of antibiotics in soil and their impact on plant pathogens. Can J Plant Pathol 39:267–274
van Loon LC, Bakker PAHM, Pieterse CMJ (1998) Systemic resistance induced by rhizosphere bacteria. Annu Rev Phytopathol 36:453–483
Varkey S, Anith KN, Narayana R, Aswini S (2018) A consortium of rhizobacteria and fungal endophyte suppress the root-knot nematode parasite in tomato. Rhizosphere 5:38–42
Yashaswini MS, Nysanth NS, Anith KN (2021) Endospore-forming bacterial endophytes from Amaranthus spp. improve plant growth and suppress leaf blight (Rhizoctonia solani Kühn) disease of Amaranthus tricolor L. Rhizosphere 19:100387
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The authors are grateful to Kerala Agricultural University for providing research facilities.
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KNA: conceptualization and designing of the experiments, writing—original draft, reviewing and editing. NSN and SLS: experimentation, data collection and compilation. CN: data compilation and writing—reviewing and editing.
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Nysanth, N.S., Sivapriya, S.L., Natarajan, C. et al. Novel in vitro methods for simultaneous screening of two antagonistic bacteria against multiple fungal phytopathogens in a single agar plate. 3 Biotech 12, 140 (2022). https://doi.org/10.1007/s13205-022-03205-3
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DOI: https://doi.org/10.1007/s13205-022-03205-3