Skip to main content
SpringerLink
Log in

Cloning and development of pathotype-specific SCAR marker associated with Sclerospora graminicola isolates from pearl millet

  • Published:
Australasian Plant Pathology Aims and scope Submit manuscript

Abstract

Downy mildew pathogen of pearl millet in India is associated with the spread of the highly virulent Sclerospora graminicola pathotype-1. Twenty-seven S. graminicola isolates were screened using 20 inter simple sequence repeats (ISSR). Dinucleotide repeat primer [17898A-(CA)6 AC] amplified a ∼600 bp fragment specific to five isolates of pathotype-1 (Sg 048, Sg 153, Sg 212, DM-11 and DM-90). The ISSR fragment linked with pathotype-1 was cloned successfully and sequenced. To convert ISSR fragments into pathotype-specific sequence characterised amplified region (SCAR) markers, PCR primers were designed using a sequence of the cloned DNA fragment. PCR amplification using SCAR primer pair (UOM3-Sg-Path1-F/R) amplified a single 284 bp band only in isolates of S. graminicola pathotype-1. This SCARprimer pair did not amplify the 284 bp product from the other five S. graminicola pathotypes or a negative control, which demonstrates primer specificity for pathotype-1. The SCAR primer pair (UOM3-Sg-Path1-F/R) obtained in this study will provide a valuable tool for rapid identification and specific detection of S. graminicola pathotype-1.

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

Similar content being viewed by others

References

  • Albani MC, Battey NH, Wilkinson MJ (2004) The development of ISSR-derived SCAR markers around the seasonal flowering locus (SFL) in Fragaria vesca. Theoretical and Applied Genetics 109, 571–579. doi: 10.1007/s00122-004-1654-4

    Article  CAS  PubMed  Google Scholar 

  • Amemiya Y, Usami T, Shishido M (2000) Specificity of genomic DNA fragments cloned from an isolate of Verticillium dahliae pathogenic to tomato. Soil Microorganisms 54, 149–157.

    Google Scholar 

  • Amruthesh KN (2000) Investigations on pathogen variability, host resistance and durability in pearl millet downy mildew interaction. PhD Thesis, University of Mysore, Mysore, India.

    Google Scholar 

  • Amruthesh KN, Geetha NP, Jørgensen HJL, de Neergaard E, Shetty HS (2005) Unsaturated fatty acids from zoospores of Sclerospora graminicola induce downy mildew resistance in pearl millet. European Journal of Plant Pathology 111, 125–127. doi: 10.1007/s10658-004-1590-9

    Article  CAS  Google Scholar 

  • Bhat SS (1973) Investigations on the biology and control of Sclerospora graminicola on Bajra. PhD Thesis, University of Mysore. Mysore, India.

    Google Scholar 

  • Geetha NP, Amruthesh KN, Shetty HS (2002) Determination of cellular fatty acid components in five pathotypes of Sclerospora graminicola, the downy mildew pathogen of Pearl millet. Journal of Mycology and Plant Pathology 32, 345–353.

    CAS  Google Scholar 

  • Gold J, Harder D, Smith FT, Aung T, Procunier J (1999) Development of a molecular marker for rust resistance genes Sr39 and Lr35 in wheat breeding lines. Electronic Journal of Biotechnology 2, 1–6.

    CAS  Google Scholar 

  • Henson JM, French R (1993) The polymerase chain reaction and plant disease diagnosis. Annual Review of Phytopathology 31, 81–109. doi: 10.1146/annurev.py.31.090193.000501

    Article  CAS  PubMed  Google Scholar 

  • Hess DE, Thakur RP, Hash CT, Sereme P, Magill CW (2002) Pearl millet downy mildew: problems and control strategies for the new millennium. In ‘Sorghum and Pearl Millet Diseases’. (Ed. JF Leslie) pp. 37–41. (Iowa State University Press: Ames, IA)

    Google Scholar 

  • Howarth CJ, Yadav RS (2002) Successful marker assisted selection for drought tolerance and disease resistance in pearl millet. IGER Innovations (6), 18–21.

    Google Scholar 

  • ICAR(Indian Council of Agricultural Research) (2005) All India Coordinated Pearl Millet Project Annual Report. pp. 33–37. Junagad, Gujarat, India.

  • ICAR(Indian Council of Agricultural Research) (2006) All India Coordinated Pearl Millet Project Annual Report. pp. 39–44. ICRISAT, Andrapradesh, India.

    Google Scholar 

  • Li KN, Rouse DI, Eyestone EJ, German TL (1999) The generation of specific DNA primers using random amplified polymorphic DNA and its application to Verticillium dahliae. Mycological Research 103, 1361–1368. doi: 10.1017/S0953756299008357

    Article  CAS  Google Scholar 

  • Mercado-Blanco J, Rodryguez-Jurado D, Perez-Artes E, Jimenez-Dyaz RM (2001) Detection of the nondefoliating pathotype of Verticillium dahliae in infected olive plants by nested PCR. Plant Pathology 50, 609–619. doi: 10.1046/j.1365-3059.2001.00601.x

    Article  CAS  Google Scholar 

  • Mercado-Blanco J, Rodryguez-Jurado D, Perez-Artes E, Jimenez-Dyaz RM (2002) Detection of the defoliating pathotype of Verticillium dahliae in infected olive plants by nestedPCR. European Journal of Plant Pathology 108, 1–13. doi: 10.1023/A:1013994827836

    Article  CAS  Google Scholar 

  • Michelmore RW, Pawar MN, Williams RJ (1982) Heterothallism in Sclerospora graminicola. Phytopathology 72, 1368–1372. doi: 10.1094/ Phyto-72-1368

    Article  Google Scholar 

  • Nazar RN, Hu X, Schmidt J, Culham D, Robb J (1991) Potential use of PCR amplified detection and differentiation of Verticillium wilt pathogens. Physiological and Molecular Plant Pathology 39, 1–11. doi: 10.1016/ 0885-5765(91)90027-F

    Article  CAS  Google Scholar 

  • Nene YL, Singh SD (1976) Downy mildew and ergot of pearl millet. Pest Article and News Summary 22, 366–385.

    Google Scholar 

  • Pérez-Artés E, Pedrajas GMD, Aleazar BJ, Diaz JRM (2000) Differentitation of cotton defoliation and nondefoliating pathotype of Verticillium dahliae by RAPD and specific PCR analyses. European Journal of Plant Pathology 106, 507–517. doi: 10.1023/A:1008756307969

    Article  Google Scholar 

  • Radisek S, Jakse J, Javornik B (2004) Development of pathotype-specific SCAR markers for detection of Verticillium albo-atrum isolates from Hop. Plant Disease 88, 1115–1122. doi: 10.1094/PDIS.2004.88. s10.1115

    Article  CAS  Google Scholar 

  • Sastry JG, Ramakrishna W, Sivaramakrishnan S, Thakur RP, Gupta VS, Ranjekar PK (1995) DNA fingerprinting detects genetic variability in pearl millet downy mildew pathogen (Sclerospora graminicola). Theoretical and Applied Genetics 91, 856–861. doi: 10.1007/ BF00223892

    Article  Google Scholar 

  • Sharathchandra RG, Geetha NP, Amruthesh KN, Kini KR, Sarosh BR, Shetty NP, Shetty HS (2006) Isolation and characterization of a protein elicitor from S. graminicola and elicitor mediated induction of defence responses in cultured cells of P. glaucum. Functional Plant Biology 33, 267–278. doi: 10.1071/FP05197

    Article  CAS  Google Scholar 

  • Shetty HS, Ahmad R (1981) Physiologic specialization in Sclerospora graminicola. Indian Phytopathology 34, 307–309.

    Google Scholar 

  • Shishupala S (1994) Variability in the Pearl Millet downy mildew pathogen. PhD Thesis, University of Mysore, Mysore, India.

    Google Scholar 

  • Singh SD (1995) Downy mildew of pearl millet. Plant Disease 79, 545–550.

    Article  Google Scholar 

  • Singru R, Sivaramakrishnan S, Thakur RP, Gupta VS, Ranjekar PK (2003) Detection of genetic variability in pearl millet downy mildew (Sclerospora graminicola) by AFLP. Biochemical Genetics 41, 361–374. doi: 10.1023/B:BIGI.0000007772.27724.0e

    Article  CAS  PubMed  Google Scholar 

  • Sudisha J, Ananda Kumar S, Shekar Shetty H (2008) Characterization of Downy Mildew Isolates of Sclerospora graminicola by using Differential Cultivars and Molecular Markers. Journal of Cell and Molecular Biology 7, 41–55.

    Google Scholar 

  • Thakur RP, Magill C, Sivaramakrishnan S, Hash CT, Shetty HS, Hess DE (2002) Variability in Sclerospora graminicola — the pearl millet downy mildew pathogen. In ‘Sorghum and Pearl Millet Diseases’. (Ed. JF Leslie) pp. 51–56. (Iowa State University Press: Ames, IA)

    Google Scholar 

  • Thakur RP, Rao VP, Amruthesh KN, Shetty HS, Datar VV (2003) Field surveys of pearl millet downy mildew effects of hybrids, fungicide and cropping sequence. Journal of Mycology and Plant Pathology 33, 387–394.

    Google Scholar 

  • Thakur RP, Rao VP, Wu BM, Subbarao KV, Shetty HS et al. (2004) Host resistance stability to downy mildew in pearl millet and pathogenic variability in Sclerospora graminicola. Crop Protection (Guildford, Surrey) 23, 901–908. doi: 10.1016/j.cropro.2004.02.003

    Article  Google Scholar 

  • Travensolo R, Ciapina LP, Lemoa EGM (2005) Development of a SCAR marker for Pierce’s disease strains of Xylella fastidiosa. Fitopatologia Brasileira 30, 115–120. doi: 10.1590/S0100-41582005000200002

    Article  Google Scholar 

  • Usami T, Abiko M, Shishido M, Amemiya Y (2002) Specific Detection of Tomato Pathotype of Verticillium dahliae by PCR Assays. Journal of General Plant Pathology 68, 134–140. doi: 10.1007/PL00013066

    Article  CAS  Google Scholar 

  • Usami T, Ishigaki S, Takashina H, Matsubara Y, Amemiya Y (2007) Cloning of DNA fragments specific to the pathotype and race of Verticillium dahliae. Journal of General Plant Pathology 73, 89–95. doi: 10.1007/ s10327-006-0334-4

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Downy Mildew Research Laboratory, Department of Studies in Applied Botany, Seed Pathology and Biotechnology, University of Mysore, Manasagangotri, 570 006, Mysore, Karnataka, India

    J. Sudisha, S. R. Niranjana & H. Shekar Shetty

  2. Microbiology and Cell Biology Division, Indian Institute of Science (IISc), 560 012, Bangalore, Karnataka, India

    S. Ananda Kumar

  3. Department of Plant Biology, Royal Veterinary and Agricultural University, Thorvaldsensvej 40, 1871 Frederiksberg C, Copenhagen, Denmark

    N. P. Shetty

Authors
  1. J. Sudisha
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. Ananda Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S. R. Niranjana
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. N. P. Shetty
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. H. Shekar Shetty
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to H. Shekar Shetty.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sudisha, J., Kumar, S.A., Niranjana, S.R. et al. Cloning and development of pathotype-specific SCAR marker associated with Sclerospora graminicola isolates from pearl millet. Australasian Plant Pathology 38, 216–221 (2009). https://doi.org/10.1071/AP08098

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1071/AP08098

Additional keywords

Search

Navigation