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Current IssueEditorial BoardOnline First ArticlesArchive

volume 34 issue 3 (september 2014) : 171 - 176,   Doi: 10.5958/0976-0547.2014.00996.3

CHARACTERIZATION OF DOWNY MILDEW RESISTANT AND SUSCEPTIBLE GENOTYPES OF PEARL MILLET (PENNISETUM GLAUCUM L.) USING SDS-PAGE AND RAPD MARKERS

S
Sarang S. Sapre*
A
Abhishek Singh
G
Gautam M. Saripalli
V
Vishal R. Patil
J
J. G. Talati
1B.A. College of Agriculture, Anand Agricultural University, Anand-388 110, India
Cite article:- Sapre* S. Sarang, Singh Abhishek, Saripalli M. Gautam, Patil R. Vishal, Talati G. J. (2025). CHARACTERIZATION OF DOWNY MILDEW RESISTANT AND SUSCEPTIBLE GENOTYPES OF PEARL MILLET (PENNISETUM GLAUCUM L.) USING SDS-PAGE AND RAPD MARKERS. Agricultural Science Digest. 34(3): 171 - 176. doi: 10.5958/0976-0547.2014.00996.3.

ABSTRACT

Protein and DNA profiling of five resistant and two susceptible pearl millet genotypes to downy mildew were investigated. Characterization of total seed proteins using SDS-PAGE was performed. The resistant genotypes had more number of bands (9-14) whereas; susceptible genotypes had fewer bands (4-6). Two bands having Rm values 0.27 and 0.36 were specifically present in all the five resistant genotypes, and were absent in both the susceptible genotypes. The PCR amplification was performed using 120 oligonucleotide decamer primers of arbitrary sequence; however only 14 oligonulcleotide primers yielding more than six PCR fragments were selected. Five PCR fragments obtained in resistant genotypes with OPB 18, OPE 12, OPI 1, OPI 14 and OPK 10 reactions respectively showed its possible linkage with downy mildew resistance gene. The result of protein and DNA profiling suggests suitability of this technique for screening of different pearl millet genotypes against S. graminicola. The development of SCAR can aid in rapid and reliable screening.

REFERENCES

  1. Anonymous (2009). In: Agricultural Statistics at a Glance, Directorate of Economics and Statistics, Department of Agriculture and co-operation, Ministry of Agriculture, Government of India, pp. 93-95.
  2. Bardakci, F. (2001). Randomly amplified polymorphic DNA (RAPD) markers. Turk. J. Biol. 25: 185-196.
  3. Belkhadir, Y., Subramaniam, R. and Dangi, J. L. (2004). Plant disease resistance protein signaling: NBS-LRR proteins and their partners. Curr. Opin. Plant Biol. 7: 391–9.
  4. Ghafoor, A., Ahmed, Z., Qureshi, A. S. and Bashir, M. (2002). Genetic relationship in Vigna mungo (L.) Hepper and V. radiate (L.) R. Wilczek based on morphological traits and SDS-PAGE. Euphytica., 123(3): 367-378.
  5. Gogoi, R., Malathi, K., Srinivasan, K. and Singh, D. V. (2005). Differentiation of karnal bunt resistant and susceptible cultivars of wheat using RAPD. Indian Phytopath., 58(2): 157-162.
  6. Govindaraj, M., Selvi, B., Prabhu, D. A. and Rajarathinam, S. (2009). Genetic diversity analysis of pearl millet accessions using molecular markers. African journal of Biotechnology., 8(22): 6046-6052.
  7. Iqbal, S. M., Ghafoor, A. and Ayub, N. (2005). Relationship between SDS-PAGE markers and Ascochyta blight in chickpea. Pak. J. Bot., 37(1): 87-95.
  8. Kalpana., Reddy, N., Kumar, A. M., Rajanna, M. D., Rahhiman, A. and Gowda, B. (2009). Evaluation of maize genotypes against downy mildew and characterization of soluble seed proteins by SDS-PAGE. Archives of phytopathology and plant protection.; 142(12): 1126-1131.
  9. Martin, G. B., Bogdanove, A. J. and Sessa, G. (2003). Understanding the functions of plant disease resistance proteins. Ann. Rev. Plant Biol., 54: 23-61.
  10. Mondal, S., Ghosh, S. and Badigannavar, A. M. (2005). RAPD polymorphism among groundnut genotypes differing in disease reaction to late leaf spot and rust. IAN., 25: 27-30.
  11. Rohlf, F.J. (1998). NTSYS-pc Numerical Taxonomy and Multivariate Analysis System, Version 2.0, Exeter Software, Setauket, New York.
  12. Sadasivam, S. and Manickam, A. (2008). Biochemical methods. New Age International (P) Limited Publishers, New Delhi.
  13. Sambrook and Russel (2001). Molecular Cloning. A laboratory Manual (3rd edition). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
  14. Shuaib, M., Zeb, A., Ali, Z., Ali, W., Ahmed, T. and Khan, I. (2007). Characterization of wheat varieties by SDS-PAGE electrophoresis. African journal of Biotechnology., 6(5): 497-500.
  15. Singh, S. D. (1995). Downy mildews of pearl millet. Plant Disease., 79(6): 545-550.
  16. Zidani, S., Ferchichi, A. and Chaieb, M. (2005). DNA extraction method from pearl millet (Pennisetum glaucum) leaves. African Journal of Biotechnology., 4(8): 862-866.
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    volume 34 issue 3 (september 2014) : 171 - 176,   Doi: 10.5958/0976-0547.2014.00996.3

    CHARACTERIZATION OF DOWNY MILDEW RESISTANT AND SUSCEPTIBLE GENOTYPES OF PEARL MILLET (PENNISETUM GLAUCUM L.) USING SDS-PAGE AND RAPD MARKERS

    S
    Sarang S. Sapre*
    A
    Abhishek Singh
    G
    Gautam M. Saripalli
    V
    Vishal R. Patil
    J
    J. G. Talati
    1B.A. College of Agriculture, Anand Agricultural University, Anand-388 110, India
    Cite article:- Sapre* S. Sarang, Singh Abhishek, Saripalli M. Gautam, Patil R. Vishal, Talati G. J. (2025). CHARACTERIZATION OF DOWNY MILDEW RESISTANT AND SUSCEPTIBLE GENOTYPES OF PEARL MILLET (PENNISETUM GLAUCUM L.) USING SDS-PAGE AND RAPD MARKERS. Agricultural Science Digest. 34(3): 171 - 176. doi: 10.5958/0976-0547.2014.00996.3.

    ABSTRACT

    Protein and DNA profiling of five resistant and two susceptible pearl millet genotypes to downy mildew were investigated. Characterization of total seed proteins using SDS-PAGE was performed. The resistant genotypes had more number of bands (9-14) whereas; susceptible genotypes had fewer bands (4-6). Two bands having Rm values 0.27 and 0.36 were specifically present in all the five resistant genotypes, and were absent in both the susceptible genotypes. The PCR amplification was performed using 120 oligonucleotide decamer primers of arbitrary sequence; however only 14 oligonulcleotide primers yielding more than six PCR fragments were selected. Five PCR fragments obtained in resistant genotypes with OPB 18, OPE 12, OPI 1, OPI 14 and OPK 10 reactions respectively showed its possible linkage with downy mildew resistance gene. The result of protein and DNA profiling suggests suitability of this technique for screening of different pearl millet genotypes against S. graminicola. The development of SCAR can aid in rapid and reliable screening.

    REFERENCES

    1. Anonymous (2009). In: Agricultural Statistics at a Glance, Directorate of Economics and Statistics, Department of Agriculture and co-operation, Ministry of Agriculture, Government of India, pp. 93-95.
    2. Bardakci, F. (2001). Randomly amplified polymorphic DNA (RAPD) markers. Turk. J. Biol. 25: 185-196.
    3. Belkhadir, Y., Subramaniam, R. and Dangi, J. L. (2004). Plant disease resistance protein signaling: NBS-LRR proteins and their partners. Curr. Opin. Plant Biol. 7: 391–9.
    4. Ghafoor, A., Ahmed, Z., Qureshi, A. S. and Bashir, M. (2002). Genetic relationship in Vigna mungo (L.) Hepper and V. radiate (L.) R. Wilczek based on morphological traits and SDS-PAGE. Euphytica., 123(3): 367-378.
    5. Gogoi, R., Malathi, K., Srinivasan, K. and Singh, D. V. (2005). Differentiation of karnal bunt resistant and susceptible cultivars of wheat using RAPD. Indian Phytopath., 58(2): 157-162.
    6. Govindaraj, M., Selvi, B., Prabhu, D. A. and Rajarathinam, S. (2009). Genetic diversity analysis of pearl millet accessions using molecular markers. African journal of Biotechnology., 8(22): 6046-6052.
    7. Iqbal, S. M., Ghafoor, A. and Ayub, N. (2005). Relationship between SDS-PAGE markers and Ascochyta blight in chickpea. Pak. J. Bot., 37(1): 87-95.
    8. Kalpana., Reddy, N., Kumar, A. M., Rajanna, M. D., Rahhiman, A. and Gowda, B. (2009). Evaluation of maize genotypes against downy mildew and characterization of soluble seed proteins by SDS-PAGE. Archives of phytopathology and plant protection.; 142(12): 1126-1131.
    9. Martin, G. B., Bogdanove, A. J. and Sessa, G. (2003). Understanding the functions of plant disease resistance proteins. Ann. Rev. Plant Biol., 54: 23-61.
    10. Mondal, S., Ghosh, S. and Badigannavar, A. M. (2005). RAPD polymorphism among groundnut genotypes differing in disease reaction to late leaf spot and rust. IAN., 25: 27-30.
    11. Rohlf, F.J. (1998). NTSYS-pc Numerical Taxonomy and Multivariate Analysis System, Version 2.0, Exeter Software, Setauket, New York.
    12. Sadasivam, S. and Manickam, A. (2008). Biochemical methods. New Age International (P) Limited Publishers, New Delhi.
    13. Sambrook and Russel (2001). Molecular Cloning. A laboratory Manual (3rd edition). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
    14. Shuaib, M., Zeb, A., Ali, Z., Ali, W., Ahmed, T. and Khan, I. (2007). Characterization of wheat varieties by SDS-PAGE electrophoresis. African journal of Biotechnology., 6(5): 497-500.
    15. Singh, S. D. (1995). Downy mildews of pearl millet. Plant Disease., 79(6): 545-550.
    16. Zidani, S., Ferchichi, A. and Chaieb, M. (2005). DNA extraction method from pearl millet (Pennisetum glaucum) leaves. African Journal of Biotechnology., 4(8): 862-866.
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