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Research Article

volume 52 issue 5 (october 2018) : 489-496,   Doi: 10.18805/IJARe.A-5038

Over expression of stress responsive Pennisetum glaucum 47 helicase (PG47) improves stress tolerance in groundnut (Arachis hypogaea L)

S
Sujata Mahantesh
R
Ramesh Babu H.N
K
Kirankumar Ghanti
T
T.G. Prasad
M
Malireddy. K. Reddy
P
P.C. Raddy
1Department of Crop Physiology, University of Agricultural Sciences, Gandhi Krishi Vignana Kendra, Bangalore-560 056, Karnataka, India.
Cite article:- Mahantesh Sujata, H.N Babu Ramesh, Ghanti Kirankumar, Prasad T.G., Reddy K. Malireddy., Raddy P.C. (2018). Over expression of stress responsive Pennisetum glaucum 47 helicase (PG47) improves stress tolerance in groundnut (Arachis hypogaea L). Indian Journal of Agricultural Research. 52(5): 489-496. doi: 10.18805/IJARe.A-5038.

ABSTRACT

RNA helicases function as molecular motors that rearrange RNA secondary structure, potentially performing roles in any cellular process involving RNA metabolism in an ATP-dependent manner and play an important role in protein synthesis. Pennisetum glaucum 47 (PG47) RNA helicase overexpressed in groundnut (Arachis hypogaea L.) cultivar GPBD-4 improved drought tolerance. The transgenics plants were confirmed for presence, expression and stable integration by Kanamycin screening, genomic DNA PCR, RT-PCR and Southern analyses respectively.  In T3 generation, the promising transgenic events were identified based on stress tolerance and improvement in productivity. The transgenic events showed enhanced stay-green phenotype and increased chlorophyll stability under drought stress. The transgenics also showed reduced chlorophyll retardation under NaCl, PEG and etherel-induced stress conditions. Transgenic plants showed increased yield than wild type under stress conditions. Results suggested that PG47 RNA helicase contributing for enhanced drought-adaptive traits and improved productivity under water-limited conditions.

REFERENCES

  1. Amin M., Elias S. M., Hossain A., Ferdousi A., Rahman M. S., Tuteja N., et al. (2012). Over-expression of a DEAD-box helicase, PDH45, confers both seedling and reproductive stage salinity tolerance to rice (Oryza sativa L.). Mol. Breed. 30: 345–354.
  2. Arnon, D., L. (1949) Copper enzymes in isolated chloroplasts poly phenoloxidase in BETA vulgaris. 24(1): 1–15
  3. De, L.Cruz, J. (1999) Unwinding RNA in Saccharomyces cerevisiae: DEAD-box proteins and related families. Trends Biochem Sci 24 (5):192-8.
  4. Dunand-Sauthier, I., Walker, C. A., Narasimhan, J., Pearce, A.K. Wek, R.C. Humphrey,T.C. Walker ,C.A. Narasimhan, J. (2005) Stress-activated protein kinase pathway functions to support protein synthesis and translational adaptation in response to environmental stress in fission yeast. Eukaryot. Cell 4:1785–1793.
  5. FAOSTAT (2014). FAO Statistics Division 2014. Available online at: http://faostat.fao.org/
  6. Govind,G.,Harshavardhan,V.T.,Patricia,J.K.,Dhanalakshmi,R.,Senthilkumar,M.,Sreenivasulu,N., Udayakumar, M (2009) Identification and functional validation of a unique set of drought induced genes preferentially expressed in response to gradual water stress in peanut. Mol Genet Genomics 281:591–605.
  7. Hiscox, J.D., Israelstam, G.F. (1979) Different methods of chlorophyll extraction. Journal of Botany 57: 332.
  8. Hunseung, K., Su, J.P., Kyung, J.K. (2013) Plant RNA chaperones in stress response. Trends Plant Sci 18 (2) : 100-106.
  9. Kiran, G. S., Sujata, K.G., Srinath, R.M. (2009) The effect of phenyl acetic acid on shoot bud induction, elongation and rooting of chickpea. Biol. Plant 53: 779-783.
  10. Levitt, J. (1980) Responses of Plant to Environmental Stress: Water, Radiation, Salt and Other Stresses. Academic Press, New York , 365.
  11. Manjulatha, M., Sreevathsa, R., Kumar, A.M., Sudhakar, C., Prasad, T.G., Tuteja, N., Udayakumar,M. (2014) Overexpression of a pea DNA helicase (PDH45) in peanut (Arachis hypogaea L.) confers improvement of cellular level tolerance and productivity under drought stress. Molecular Biotechnology 56 (2): 111–125.
  12. Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-497.
  13. Nooden, L.D., Guiamet, J.J., John, I. (1997) Senescence mechanisms introduction. Physiol Plant 101:746-753
  14. Pham, X.H., Reddy, M.K., Ehtesham, N.Z., Matta, B., Tuteja, N (2000). A DNA helicase from Pisum sativum is homologous totranslation initiation factor and stimulates topoisomerase I activity. Plant J 24: 219–229.
  15. Rivero, M.R., Shulev,V., Blumwald, E. (2009) Cytokynin-dependent photorespiration and the protection of photosynthesis during water deûcit. Plant Physiology 50:1530–1540.
  16. Rocak, S.,Linder, P (2004) DEAD-box proteins: the driving forces behind RNA metabolism. Nature Rev. Mol Cell Biol 5: 232–241.
  17. Sambrook, J, Fritish, E.F, Maniatis, T. (1989) Molecular Cloning: A Molecular Manual. 2nd Ed. - Cold Spring Harbor Laboratory Press. Cold Spring Harbor - New York.
  18. Santosh, R.B.R.T, Vijaya, N.J, Sudhakar, R.P, Reddy, M.K, Mallikarjuna, G. (2017) Expression of Pennisetum glaucum Eukaryotic Translational Initiation Factor 4A (PgeIF4A) Confers Improved Drought, Salinity, and Oxidative Stress Tolerance in Groundnut. Front Plant Sci 8:453.
  19. Sarkar, A., Markus, P., Thoms, M., Beckmann, R. (2016) Ribosome-stalk biogenesis is coupled with recruitment of nuclear-export factor to the nascent 60S subunit. Nat Struct Mol Biol 23 (12):1074-1082.
  20. Sruthy, M.A., Ashwin,N.J., Divya, P., Syamaladevi, C., Appunu, M., Chakravarthi, V.R., Tuteja N, Subramonian. (2015) Introduction of pea DNA Helicase 45 into sugarcane (Saccharum spp. Hybrid) enhances cell membrane thermostability and upregulation of stress-responsive genes leads to abiotic stress tolerance. Mol Biotechnol: doi/ 015-98.
  21. Tanner,N.K., Linder, P. (2001) DExD/H box RNA helicases: from generic motors to specific dissociation functions. Mol Cell: 8: 251–262.
  22. Tuteja, N.,Tuteja, R. (2004b) Unraveling DNA helicases. Motif, structure, mechanism and function. Eur J Biochem 271: 1849–1863.
  23. Zhu, J.K. (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53: 247–273.
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Indian Journal of Agricultural Research
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    Research Article

    volume 52 issue 5 (october 2018) : 489-496,   Doi: 10.18805/IJARe.A-5038

    Over expression of stress responsive Pennisetum glaucum 47 helicase (PG47) improves stress tolerance in groundnut (Arachis hypogaea L)

    S
    Sujata Mahantesh
    R
    Ramesh Babu H.N
    K
    Kirankumar Ghanti
    T
    T.G. Prasad
    M
    Malireddy. K. Reddy
    P
    P.C. Raddy
    1Department of Crop Physiology, University of Agricultural Sciences, Gandhi Krishi Vignana Kendra, Bangalore-560 056, Karnataka, India.
    Cite article:- Mahantesh Sujata, H.N Babu Ramesh, Ghanti Kirankumar, Prasad T.G., Reddy K. Malireddy., Raddy P.C. (2018). Over expression of stress responsive Pennisetum glaucum 47 helicase (PG47) improves stress tolerance in groundnut (Arachis hypogaea L). Indian Journal of Agricultural Research. 52(5): 489-496. doi: 10.18805/IJARe.A-5038.

    ABSTRACT

    RNA helicases function as molecular motors that rearrange RNA secondary structure, potentially performing roles in any cellular process involving RNA metabolism in an ATP-dependent manner and play an important role in protein synthesis. Pennisetum glaucum 47 (PG47) RNA helicase overexpressed in groundnut (Arachis hypogaea L.) cultivar GPBD-4 improved drought tolerance. The transgenics plants were confirmed for presence, expression and stable integration by Kanamycin screening, genomic DNA PCR, RT-PCR and Southern analyses respectively.  In T3 generation, the promising transgenic events were identified based on stress tolerance and improvement in productivity. The transgenic events showed enhanced stay-green phenotype and increased chlorophyll stability under drought stress. The transgenics also showed reduced chlorophyll retardation under NaCl, PEG and etherel-induced stress conditions. Transgenic plants showed increased yield than wild type under stress conditions. Results suggested that PG47 RNA helicase contributing for enhanced drought-adaptive traits and improved productivity under water-limited conditions.

    REFERENCES

    1. Amin M., Elias S. M., Hossain A., Ferdousi A., Rahman M. S., Tuteja N., et al. (2012). Over-expression of a DEAD-box helicase, PDH45, confers both seedling and reproductive stage salinity tolerance to rice (Oryza sativa L.). Mol. Breed. 30: 345–354.
    2. Arnon, D., L. (1949) Copper enzymes in isolated chloroplasts poly phenoloxidase in BETA vulgaris. 24(1): 1–15
    3. De, L.Cruz, J. (1999) Unwinding RNA in Saccharomyces cerevisiae: DEAD-box proteins and related families. Trends Biochem Sci 24 (5):192-8.
    4. Dunand-Sauthier, I., Walker, C. A., Narasimhan, J., Pearce, A.K. Wek, R.C. Humphrey,T.C. Walker ,C.A. Narasimhan, J. (2005) Stress-activated protein kinase pathway functions to support protein synthesis and translational adaptation in response to environmental stress in fission yeast. Eukaryot. Cell 4:1785–1793.
    5. FAOSTAT (2014). FAO Statistics Division 2014. Available online at: http://faostat.fao.org/
    6. Govind,G.,Harshavardhan,V.T.,Patricia,J.K.,Dhanalakshmi,R.,Senthilkumar,M.,Sreenivasulu,N., Udayakumar, M (2009) Identification and functional validation of a unique set of drought induced genes preferentially expressed in response to gradual water stress in peanut. Mol Genet Genomics 281:591–605.
    7. Hiscox, J.D., Israelstam, G.F. (1979) Different methods of chlorophyll extraction. Journal of Botany 57: 332.
    8. Hunseung, K., Su, J.P., Kyung, J.K. (2013) Plant RNA chaperones in stress response. Trends Plant Sci 18 (2) : 100-106.
    9. Kiran, G. S., Sujata, K.G., Srinath, R.M. (2009) The effect of phenyl acetic acid on shoot bud induction, elongation and rooting of chickpea. Biol. Plant 53: 779-783.
    10. Levitt, J. (1980) Responses of Plant to Environmental Stress: Water, Radiation, Salt and Other Stresses. Academic Press, New York , 365.
    11. Manjulatha, M., Sreevathsa, R., Kumar, A.M., Sudhakar, C., Prasad, T.G., Tuteja, N., Udayakumar,M. (2014) Overexpression of a pea DNA helicase (PDH45) in peanut (Arachis hypogaea L.) confers improvement of cellular level tolerance and productivity under drought stress. Molecular Biotechnology 56 (2): 111–125.
    12. Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-497.
    13. Nooden, L.D., Guiamet, J.J., John, I. (1997) Senescence mechanisms introduction. Physiol Plant 101:746-753
    14. Pham, X.H., Reddy, M.K., Ehtesham, N.Z., Matta, B., Tuteja, N (2000). A DNA helicase from Pisum sativum is homologous totranslation initiation factor and stimulates topoisomerase I activity. Plant J 24: 219–229.
    15. Rivero, M.R., Shulev,V., Blumwald, E. (2009) Cytokynin-dependent photorespiration and the protection of photosynthesis during water deûcit. Plant Physiology 50:1530–1540.
    16. Rocak, S.,Linder, P (2004) DEAD-box proteins: the driving forces behind RNA metabolism. Nature Rev. Mol Cell Biol 5: 232–241.
    17. Sambrook, J, Fritish, E.F, Maniatis, T. (1989) Molecular Cloning: A Molecular Manual. 2nd Ed. - Cold Spring Harbor Laboratory Press. Cold Spring Harbor - New York.
    18. Santosh, R.B.R.T, Vijaya, N.J, Sudhakar, R.P, Reddy, M.K, Mallikarjuna, G. (2017) Expression of Pennisetum glaucum Eukaryotic Translational Initiation Factor 4A (PgeIF4A) Confers Improved Drought, Salinity, and Oxidative Stress Tolerance in Groundnut. Front Plant Sci 8:453.
    19. Sarkar, A., Markus, P., Thoms, M., Beckmann, R. (2016) Ribosome-stalk biogenesis is coupled with recruitment of nuclear-export factor to the nascent 60S subunit. Nat Struct Mol Biol 23 (12):1074-1082.
    20. Sruthy, M.A., Ashwin,N.J., Divya, P., Syamaladevi, C., Appunu, M., Chakravarthi, V.R., Tuteja N, Subramonian. (2015) Introduction of pea DNA Helicase 45 into sugarcane (Saccharum spp. Hybrid) enhances cell membrane thermostability and upregulation of stress-responsive genes leads to abiotic stress tolerance. Mol Biotechnol: doi/ 015-98.
    21. Tanner,N.K., Linder, P. (2001) DExD/H box RNA helicases: from generic motors to specific dissociation functions. Mol Cell: 8: 251–262.
    22. Tuteja, N.,Tuteja, R. (2004b) Unraveling DNA helicases. Motif, structure, mechanism and function. Eur J Biochem 271: 1849–1863.
    23. Zhu, J.K. (2002) Salt and drought stress signal transduction in plants. Annu Rev Plant Biol 53: 247–273.
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