Stem rot of groundnut caused by the soilborne pathogen Sclerotium rolfsii can cause significant yield losses. Biological control of stem rot using actinomycetes is a viable alternative to existing fungicidal management. Though actinomycetes are prolific antibiotic producers, reports pertaining to their use in groundnut disease management are limited. Here, actinomycetes were isolated from groundnut rhizospheric soils and screened for antagonism against S. rolfsii through a dual culture assay. Culture filtrates and crude extracts of the potential candidates were screened further for extracellular antifungal activity and characterized for biocontrol and plant-growth-promoting traits. A promising candidate was tested under greenhouse conditions as whole organism as well as crude extracts. Isolate RP1A-12 exhibited high antagonism against S. rolfsii in dual culture assay (69 % inhibition), culture filtrate assay (78–100 % inhibition at various concentrations) and crude extract assay (100 % inhibition with 1 % crude extracts). Moreover, germination of sclerotia of the test pathogen was inhibited with 1 % crude extracts. Strain RP1A-12 produced hydrogen cyanide, lipase, siderophores and indole acetic acid. Oxalic acid production by S. rolfsii was also inhibited by crude extracts of RP1A-12. In greenhouse studies, RP1A-12 reduced stem rot severity. Overall, our results suggest that isolate RP1A-12 has potential biocontrol capabilities against stem rot pathogen. Molecular characterization based on 16S rRNA gene sequencing of RP1A-12 identified it as a species of Streptomyces, closely related to S. flocculus.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Ahmad F, Ahmad I, Khan MS (2008) Screening of free-living rhizospheric bacteria for their multiple plant growth promoting activities. Microbiol Res 163:173–181
Aldesuquy HS, Mansour FA, Abo-Hamed SA (1998) Effect of the culture filtrates of Streptomyces on growth and productivity of wheat plants. Folia Microbiol 43:465–470
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Anjaiah V, Cornelis P, Koedam N (2003) Effect of genotype and root colonization in biological control of fusarium wilts in pigeonpea and chickpea by Pseudomonas aeruginosa PNA1. Can J Microbiol 49:85–91
Bhattacharya A, Chandra S, Barik S (2009) Lipase and protease producing microbes from the environment of sugar beet field. Ind J Agric Biochem 22:26–30
Bouizgarne B (2013) Bacteria for plant growth promotion and disease management. In: Maheshwari DK (ed) Bacteria in agrobiology: disease management. Springer, Heidelberg, pp 15–47
Bowen KL, Hagan AK, Weeks R (1992) Seven years of Sclerotium rolfsii in peanut fields: yield losses and means of minimization. Plant Dis 76:982–985
Cessna SG, Sears VE, Dickman MB, Low PS (2000) Oxalic acid, a pathogenicity factor for Sclerotinia sclerotiorum, suppresses the oxidative burst of the host plant. Plant Cell 12:2191–2200
Chaiharn M, Chunhaleuchanon S, Lumyong S (2009) Screening siderophore producing bacteria as potential biological control agent for fungal rice pathogens in Thailand. World J Microbiol Biotechnol 25:1919–1928
Crawford DL, Lynch JM, Whipps JM, Ousley MA (1993) Isolation and characterization of actinomycete antagonists of a fungal root pathogen. Appl Environ Microbiol 59:3899–3905
Dennis C, Webster J (1971) Antagonistic properties of species-groups of Trichoderma: III. Hyphal interaction. Trans Br Mycol Soc 57:363–369
Dey R, Pal KK, Bhatt DM, Chauhan SM (2004) Growth promotion and yield enhancement of peanut (Arachis hypogaea L.) by application of plant growth-promoting rhizobacteria. Microbiol Res 159:371–394
Doumbou CL, Salove MKH, Crawford DL, Beaulieu C (2001) Actinomycetes, promising tools to control plant diseases and to promote plant growth. Phytoprotection 82:85–102
Duijff BJ, Meijer JW, Bakker PAHM, Schippers B (1993) Siderophore-mediated competition for iron and induced resistance in the suppression of fusarium wilt of carnation by fluorescent Pseudomonas spp. Neth J Plant Pathol 99:277–289
El-Tarabily KA, Soliman MH, Nassar AH, Al-Hassani HA, Sivasithamparam K, McKenna F, Hardy GS (2000) Biological control of Sclerotinia minor using a chitinolytic bacterium and actinomycetes. Plant Pathol 49:573–583
Endo A, Kakiki K, Misato T (1970) Mechanism of action of the antifungal agent polyoxin D. J Bacteriol 104:189–196
Gawande SP, Borkar SG, Chimote VP (2013) Variation in growth and oxalic acid production by different crop isolates of Sclerotium rolfsii Sacc. J Mycopathol Res 51:95–100
Godoy G, Steadman JR, Dickman MB, Dam R (1990) Use of mutants to demonstrate the role of oxalic acid in pathogenicity of Sclerotinia sclerotiorum on Phaseolus vulgaris. Physiol Mol Plant Pathol 37:179–191
Gopalakrishnan S, Pande S, Sharma M, Humayun P, Kiran BK, Sandeep D, Vidya MS, Deepthi K, Rupela O (2011) Evaluation of actinomycete isolates obtained from herbal vermicompost for the biological control of Fusarium wilt of chickpea. Crop Prot 30:1070–1078
Gopalakrishnan S, Srinivas V, Vidya, Rathore A (2013) Plant growth-promoting activities of Streptomyces spp. in sorghum and rice. SpringerPlus 2:574
Gyaneshwar P, Naresh Kumar G, Parekh LJ (1998) Effect of buffering on the phosphate-solubilizing ability of microorganisms. World J Microbiol Biotechnol 14:669–673
Hagan AK, Weeks JR, Bowen K (1991) Effects of application timing and method on control of southern stem rot of peanut with foliar-applied fungicides. Peanut Sci 18:47–50
Hagan AK, Campbell HL, Bowen KL, Wells L, Goodman R (2010) Managing early leaf spot and stem rot with reduced fungicide inputs on disease-resistant peanut cultivars. Peanut Sci 37:129–136
Hirano S, Nagao N (1988) An improved method for the preparation of colloidal chitin by using methanesulfonic acid. Agric Biol Chem 52:2111–2112
Jog R, Pandya M, Nareshkumar G, Rajkumar S (2014) Mechanism of phosphate solubilization and antifungal activity of Streptomyces spp. isolated from wheat roots and rhizosphere and their application in improving plant growth. Microbiology 160:778–788
Kortemaa H, Rita H, Haahtela K, Smolander A (1994) Root-colonization ability of antagonistic Streptomyces griseoviridis. Plant Soil 163:77–83
Le CN (2011) Diversity and biological control of Sclerotium rolfsii, causal agent of stem rot of groundnut. PhD thesis, Wageningen University, Wageningen
Lorck H (1948) Production of hydrocyanic acid by bacteria. Physiol Plant 1:142–146
Macagnan D, Romeiro RS, Pomella AWV, deSouza JT (2008) Production of lytic enzymes and siderophores, and inhibition of germination of basidiospores of Moniliophthora (ex Crinipellis) perniciosa by phylloplane actinomycetes. Biol Control 47:309–314
Mahadevan A, Sridhar R (1986) Methods in physiological plant pathology, 3rd edn. Sivakami, Madras, p 182
Mayee CD, Datar VV (1988) Diseases of groundnut in the tropics. Rev Trop Plant Pathol 5:85–118
Nene YL, Thapliyal PN (1993) Fungicides in plant disease control, 3rd edn. Oxford and IBH Publishing, New Delhi
Pande S, Rao JN (2000) Changing scenario of groundnut diseases in Andhra Pradesh, Karnataka and Tamil Nadu states of India. Int Arachis Newsl 20:42–44
Paramasivan M, Mohan S, Muthukrishnan N, Chandrasekaran A (2013) Degradation of oxalic acid (OA) producing Sclerotium rolfsii (Sacc.) by organic biocides. Arch Phytopath Plant Prot 46:357–363
Patten CL, Glick BR (1996) Bacterial biosynthesis of indole-3-acetic acid. Can J Microbiol 42:207–220
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56
Seong CN, Choi JH, Baik KS (2001) An improved selective isolation of rare actinomycetes from forest soil. J Microbiol 39:17–23
Shokes FM, Róźalski K, Gorbet DW, Brenneman TB, Berger DA (1996) Techniques for inoculation of peanut with Sclerotium rolfsii in the greenhouse and field. Peanut Sci 23:124–128
Sterner O (2012) Isolation of microbial natural products. In: Sarker SD, Nahar L (eds) Natural products isolation. Methods in molecular biology, vol. 864. Springer Protocols, Heidelberg pp 393–413
Stockwell VO, Duffy B (2012) Use of antibiotics in plant agriculture. Rev Sci Tech Off Int Epiz 31:199–210
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA 6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729
Thompson JD, Gibsom TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 24:4876–4882
Tokala RK, Strap JL, Jung CM, Crawford DL, Salove MH, Deobald LA, Bailey JF, Morra MJ (2002) Novel plant–microbe rhizosphere interaction involving Streptomyces lydicus WYEC108 and the pea plant (Pisum sativum). Appl Environ Microbiol 68:2161–2171
Trejo-Estrada SR, Paszczynski A, Crawford DL (1998) Antibiotics and enzymes produced by the biocontrol agent Streptomyces violaceusniger YCED-9. J Ind Microbiol Biotechnol 21:81–90
Valois D, Fayad K, Barasubiye T, Garon M, Dery C, Brzezinski R, Beaulieu C (1996) Glucanolytic actinomycetes antagonistic to Phytophthora fragariae var. rubi, the causal agent of raspberry root rot. Appl Environ Microbiol 62:1630–1635
Vidhyasekaran P, Muthamilan M (1995) Development of formulations of Pseudomonas fluorescens for control of chickpea wilt. Plant Dis 79:782–786
Voisard C, Keel C, Haas D, Défago G (1989) Cyanide production by Pseudomonas fluorescens helps suppress black root rot of tobacco under gnotobiotic conditions. EMBO J 8:351–358
Yuan WM, Crawford DL (1995) Characterization of Streptomyces lydicus WYEC108 as a potential biocontrol agent against fungal root and seed rots. Appl Environ Microbiol 612:3119–3128
The authors are thankful to Biotechnology Industry Research Assistance Council (BIRAC), A Government of India Enterprise, for providing financial support under Biotechnology Industry Partnership Programme (BIPP) scheme (Proposal No. BT/BIPP0429/11/10). The authors are also thankful to Sri Biotech Laboratories India Pvt. Ltd, Hyderabad, India for their collaboration on this project.
Conflict of interest
The authors confirm that there are no conflicts of interest regarding any experimental data.
No laws have been violated while carrying out any of the experiments for this study.
About this article
Cite this article
Jacob, S., Sajjalaguddam, R.R., Kumar, K.V.K. et al. Assessing the prospects of Streptomyces sp. RP1A-12 in managing groundnut stem rot disease caused by Sclerotium rolfsii Sacc. J Gen Plant Pathol 82, 96–104 (2016). https://doi.org/10.1007/s10327-016-0644-0
- Stem rot
- Streptomyces sp.
- Antifungal metabolites