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Research Article

volume 53 issue 4 (august 2019) : 478-482,   Doi: 10.18805/IJARe.A-399

Changes in growth, biochemical components and antioxidant genes expression in rice seedling (Oryza sativa L.) cultivar ‘IR64’ under salt stress

M
M. Pharmawati
I
I.M.A.S. Wijaya
1Biology Department, Faculty of Mathematics and Natural Sciences, Udayana University, Kampus Bukit Jimbaran, Bali, 80361, Indonesia.
Cite article:- Pharmawati M., Wijaya I.M.A.S. (2019). Changes in growth, biochemical components and antioxidant genes expression in rice seedling (Oryza sativa L.) cultivar ‘IR64’ under salt stress. Indian Journal of Agricultural Research. 53(4): 478-482. doi: 10.18805/IJARe.A-399.

ABSTRACT

One of abiotic stresses that affects rice growth is salinity.  Plant must develop adaptation process which includes morphological, biochemical and molecular changes.  This research aimed to evaluate morpho-biochemical and molecular responses of rice ‘IR64’ to several levels of salinity stress at seedling stage.  Seedlings of ‘IR64’ were grown in a hydroponic system and treated with different levels of salinity stress (4dSm-1, 6dSm-1, and 12dSm-1) for seven days.  Responses were recorded on the final day of salt treatments. Gene expression analyses were done by semi-quantitative RT-PCR. RNA was extracted using RNase plant mini kit (Qiagen) and cDNA was synthesized using GoSript™ Reverse Transcription System (Promega). Results showed that shoot height and fresh weight decreased under salt stress.  At plants treated with salt, the chlorophyll contents were lower than that of control plants, while MDA levels were higher in salt treated plants. Semi-quantitative PCR for MnSOD1and cCu/ZnSOD1 revealed that MnSOD1 and cCu/ZnSOD1expressions increased under salt stress which indicated oxidative stress defence, with the highest expression at 4dSm-1and 6dSm-1 treatment, respectively.

REFERENCES

  1. Ali, Y., Aslam, Z., Ashraf, M.Y., Tahir, G.R. (2004). Effect of salinity on chlorophyll concentration, leaf area, yield and yield components of rice genotypes grown under saline environment. Int. J. Environ. Sci. Technol., 1: 221-225
  2. Amirjani, M.R. (2011).Effect of salinity stress on growth, sugar content, pigments and enzyme activity of rice. Int. J. Bot., 7: 73-81. 
  3. Aref, F., Rad, H.E. (2012). Physiological characterization of rice under salinity stress during vegetative and reproductive stages. Indian J. Sci. Technol., 5: 2578-2586 
  4. Arnon, D.I. (1949). Copper enzymes in isolated chloroplasts polyphenoloxidase in Beta vulgaris. Plant Physiol., 24: 1-15
  5. Baek, K.H., Skinner, D.Z. (2010). Molecular cloning and expression of sequence variantsofmanganese superoxide dismutase genes from wheat. Korean J. Environ. Agric., 29: 77-85
  6. Banumathy, S., Veni, K., Anandhababu, R., Arunachalam, P., Raveendran, M., Vanniarajan, C. (2018). Character association and stress indices for yield components in Saltol introgressed backcross inbred lines of rice (Oryza sativa L.). Indian J. Agric. Res., 52 (1): 28-33
  7. Dhanyalakshmi, K.H., Vijayalakshmi, C., Boominathan, P. (2013). Evaluation of physiological and biochemical responses of rice (Oryza sativa L.) varieties to salt stress. Indian J. Agric. Res., 47 (2): 91 - 99
  8. Endo, T., Yamamoto, S., Larrinaga, J.A., Fujiyama, H., Honna, T. (2011). Status and causes of soil salinization of irrigated agricultural lands in Southern Baja California, Mexico. Appl. Environ. Soil. Sci., 2011, Article ID 873625. DOI:10.1155/2011/873625
  9. Gill, S.S., Tuteja, N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol. Biochem., 48: 909-930.
  10. Hong, C-Y., Hsu, Y.T., Tsai, Y-C., Kao, C.H. (2007). Expression of Ascorbat peroxidase 8 in roots of rice (Oryza sativa L.) seedlings in response to NaCl. J. Exp. Bot., 58: 3273–3283
  11. Khush, G.S., Virk, P.S. (2005). IR Varieties and Their Impact, Philippines: IRRI. Manila,
  12. Kim, J-H., Chun, B.Y., Kim, J-S., Wi, S.G., Yang, D.H., Lee, C-H., Lee, M.C.(2004).Construction of gene-specific primers for various antioxidant isoenzyme genes and their expressions in rice (Oryza sativa L.) seedlings obtained from gamma-irradiated seeds. J. Photosci., 11: 115-120
  13. Kura-Hotta, M., Satoh, K., Katoh, S. (1987). Relationship between photosynthesis and chlorophyll content during leaf senescence of rice seedlings. Plant Cell Physiol., 7: 1321-1329
  14. Lutts, S., Kinet, M., Bouhamont, J. (1996). NaCl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Ann. Bot., 78:389-398
  15. Munns, R. (2005) Genes and salt tolerance: bringing them together. New Phytol., 167: 645–663
  16. Reddy, I.N.B.L., Kim, B.K., Yoon, I-S., Kim, K-H., Kwon, T-R. (2017). Salt Tolerance in Rice: Focus on Mechanisms and Approaches. Rice Sci., 24: 123-144
  17. Refli, Purwestri, Y.A. (2016).The response of antioxidant genes in rice (Oryza sativa L.) seedling Cv. Cempo Ireng under drought and salinity stresses. AIP Conference Proceedings 1744, 020047; doi: 10.1063/1.4953521
  18. Schneider, C.A., Rasband,W.S., Eliceiri, K.W. (2012). NIH image to Image J: 25 years of image analysis. Nature Methods 9: 671-675
  19. Shrivastava, P., Kumar, R. (2015). Soil salinity: A serious environmental issue and plant growth promoting bacteria as one of the tools for its alleviation. Saudi J. Biol. Sci., 22: 123-131
  20. Szabolcs, I. (1989). Salt-affected Soils. CRC Press Boca Raton, Florida: 
  21. Tanaka, Y., Hibino, T., Hayashi, Y., Tanaka, A., Kishitani, S., Takabe, T., Yokota, S. (1999).Salt tolerance of transgenic rice over-    expressing yeast mitochondrial Mn-SOD in chloroplasts. Plant Sci., 148: 131-138
  22. Tatar, Ö., Brueck, H., Gevrek, M.N., Asch, F. (2010). Physiological responses of two Turkish rice (Oryza sativa L.) varieties to salinity. Turk. J. Agric. For., 34, DOI:10.3906/tar-0908-311
  23. Wang, W., Vinocur, B., Altman, A. (2003). Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218: 1-14 
  24. Xu, L., Han, L., Huang, B. (2011). Antioxidant enzyme activities and gene expression. patterns in leaves of kentucky bluegrass in response to drought and post-drought. recovery. J. Am. Soc. Hortic. Sci., 136: 247–255 
  25. Xu, Y., Burgess, P., Huang, B. (2015). Root antioxidant mechanisms in relation to root thermo tolerance in perennial grass species contrasting in heat tolerance. PLoSONE 10(9): e0138268.doi:10.1371/journal.pone.0138268
  26. Zhang, Z-H., Liu, Q., Song, H-X., Rong, X-M., Ismail, A.M. (2012). Responses of different rice (Oryza sativa L.) genotypes to salt stress and relation to carbohydrate metabolism and chlorophyll content. Afr. J. Agric. Res., 7(1): 19-27. 
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Indian Journal of Agricultural Research
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    Research Article

    volume 53 issue 4 (august 2019) : 478-482,   Doi: 10.18805/IJARe.A-399

    Changes in growth, biochemical components and antioxidant genes expression in rice seedling (Oryza sativa L.) cultivar ‘IR64’ under salt stress

    M
    M. Pharmawati
    I
    I.M.A.S. Wijaya
    1Biology Department, Faculty of Mathematics and Natural Sciences, Udayana University, Kampus Bukit Jimbaran, Bali, 80361, Indonesia.
    Cite article:- Pharmawati M., Wijaya I.M.A.S. (2019). Changes in growth, biochemical components and antioxidant genes expression in rice seedling (Oryza sativa L.) cultivar ‘IR64’ under salt stress. Indian Journal of Agricultural Research. 53(4): 478-482. doi: 10.18805/IJARe.A-399.

    ABSTRACT

    One of abiotic stresses that affects rice growth is salinity.  Plant must develop adaptation process which includes morphological, biochemical and molecular changes.  This research aimed to evaluate morpho-biochemical and molecular responses of rice ‘IR64’ to several levels of salinity stress at seedling stage.  Seedlings of ‘IR64’ were grown in a hydroponic system and treated with different levels of salinity stress (4dSm-1, 6dSm-1, and 12dSm-1) for seven days.  Responses were recorded on the final day of salt treatments. Gene expression analyses were done by semi-quantitative RT-PCR. RNA was extracted using RNase plant mini kit (Qiagen) and cDNA was synthesized using GoSript™ Reverse Transcription System (Promega). Results showed that shoot height and fresh weight decreased under salt stress.  At plants treated with salt, the chlorophyll contents were lower than that of control plants, while MDA levels were higher in salt treated plants. Semi-quantitative PCR for MnSOD1and cCu/ZnSOD1 revealed that MnSOD1 and cCu/ZnSOD1expressions increased under salt stress which indicated oxidative stress defence, with the highest expression at 4dSm-1and 6dSm-1 treatment, respectively.

    REFERENCES

    1. Ali, Y., Aslam, Z., Ashraf, M.Y., Tahir, G.R. (2004). Effect of salinity on chlorophyll concentration, leaf area, yield and yield components of rice genotypes grown under saline environment. Int. J. Environ. Sci. Technol., 1: 221-225
    2. Amirjani, M.R. (2011).Effect of salinity stress on growth, sugar content, pigments and enzyme activity of rice. Int. J. Bot., 7: 73-81. 
    3. Aref, F., Rad, H.E. (2012). Physiological characterization of rice under salinity stress during vegetative and reproductive stages. Indian J. Sci. Technol., 5: 2578-2586 
    4. Arnon, D.I. (1949). Copper enzymes in isolated chloroplasts polyphenoloxidase in Beta vulgaris. Plant Physiol., 24: 1-15
    5. Baek, K.H., Skinner, D.Z. (2010). Molecular cloning and expression of sequence variantsofmanganese superoxide dismutase genes from wheat. Korean J. Environ. Agric., 29: 77-85
    6. Banumathy, S., Veni, K., Anandhababu, R., Arunachalam, P., Raveendran, M., Vanniarajan, C. (2018). Character association and stress indices for yield components in Saltol introgressed backcross inbred lines of rice (Oryza sativa L.). Indian J. Agric. Res., 52 (1): 28-33
    7. Dhanyalakshmi, K.H., Vijayalakshmi, C., Boominathan, P. (2013). Evaluation of physiological and biochemical responses of rice (Oryza sativa L.) varieties to salt stress. Indian J. Agric. Res., 47 (2): 91 - 99
    8. Endo, T., Yamamoto, S., Larrinaga, J.A., Fujiyama, H., Honna, T. (2011). Status and causes of soil salinization of irrigated agricultural lands in Southern Baja California, Mexico. Appl. Environ. Soil. Sci., 2011, Article ID 873625. DOI:10.1155/2011/873625
    9. Gill, S.S., Tuteja, N. (2010). Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol. Biochem., 48: 909-930.
    10. Hong, C-Y., Hsu, Y.T., Tsai, Y-C., Kao, C.H. (2007). Expression of Ascorbat peroxidase 8 in roots of rice (Oryza sativa L.) seedlings in response to NaCl. J. Exp. Bot., 58: 3273–3283
    11. Khush, G.S., Virk, P.S. (2005). IR Varieties and Their Impact, Philippines: IRRI. Manila,
    12. Kim, J-H., Chun, B.Y., Kim, J-S., Wi, S.G., Yang, D.H., Lee, C-H., Lee, M.C.(2004).Construction of gene-specific primers for various antioxidant isoenzyme genes and their expressions in rice (Oryza sativa L.) seedlings obtained from gamma-irradiated seeds. J. Photosci., 11: 115-120
    13. Kura-Hotta, M., Satoh, K., Katoh, S. (1987). Relationship between photosynthesis and chlorophyll content during leaf senescence of rice seedlings. Plant Cell Physiol., 7: 1321-1329
    14. Lutts, S., Kinet, M., Bouhamont, J. (1996). NaCl-induced senescence in leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Ann. Bot., 78:389-398
    15. Munns, R. (2005) Genes and salt tolerance: bringing them together. New Phytol., 167: 645–663
    16. Reddy, I.N.B.L., Kim, B.K., Yoon, I-S., Kim, K-H., Kwon, T-R. (2017). Salt Tolerance in Rice: Focus on Mechanisms and Approaches. Rice Sci., 24: 123-144
    17. Refli, Purwestri, Y.A. (2016).The response of antioxidant genes in rice (Oryza sativa L.) seedling Cv. Cempo Ireng under drought and salinity stresses. AIP Conference Proceedings 1744, 020047; doi: 10.1063/1.4953521
    18. Schneider, C.A., Rasband,W.S., Eliceiri, K.W. (2012). NIH image to Image J: 25 years of image analysis. Nature Methods 9: 671-675
    19. Shrivastava, P., Kumar, R. (2015). Soil salinity: A serious environmental issue and plant growth promoting bacteria as one of the tools for its alleviation. Saudi J. Biol. Sci., 22: 123-131
    20. Szabolcs, I. (1989). Salt-affected Soils. CRC Press Boca Raton, Florida: 
    21. Tanaka, Y., Hibino, T., Hayashi, Y., Tanaka, A., Kishitani, S., Takabe, T., Yokota, S. (1999).Salt tolerance of transgenic rice over-    expressing yeast mitochondrial Mn-SOD in chloroplasts. Plant Sci., 148: 131-138
    22. Tatar, Ö., Brueck, H., Gevrek, M.N., Asch, F. (2010). Physiological responses of two Turkish rice (Oryza sativa L.) varieties to salinity. Turk. J. Agric. For., 34, DOI:10.3906/tar-0908-311
    23. Wang, W., Vinocur, B., Altman, A. (2003). Plant responses to drought, salinity and extreme temperatures: towards genetic engineering for stress tolerance. Planta 218: 1-14 
    24. Xu, L., Han, L., Huang, B. (2011). Antioxidant enzyme activities and gene expression. patterns in leaves of kentucky bluegrass in response to drought and post-drought. recovery. J. Am. Soc. Hortic. Sci., 136: 247–255 
    25. Xu, Y., Burgess, P., Huang, B. (2015). Root antioxidant mechanisms in relation to root thermo tolerance in perennial grass species contrasting in heat tolerance. PLoSONE 10(9): e0138268.doi:10.1371/journal.pone.0138268
    26. Zhang, Z-H., Liu, Q., Song, H-X., Rong, X-M., Ismail, A.M. (2012). Responses of different rice (Oryza sativa L.) genotypes to salt stress and relation to carbohydrate metabolism and chlorophyll content. Afr. J. Agric. Res., 7(1): 19-27. 
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