Journal of Plant Sciences

| Peer-Reviewed |

An Efficient Callus Induction and Regeneration Protocol for a Drought Tolerant Rice Indica Genotype AC39020

Received: 07 August 2015    Accepted: 21 August 2015    Published: 07 September 2015
Views:       Downloads:

Share This Article

Abstract

Most of the cultivated indica rice genotypes are less amenable to genetic modifications due to their poor callus induction and regeneration potential. The prerequisite for genetic enhancement of indica rice genotypes by biotechnological approach is to develop an efficient protocol for callus induction and plant regeneration. In the present study, we established an optimized regeneration protocol for rice genotype AC39020, which is moderately drought tolerant with high root growth and biomass. To use this genotype in the crop improvement program the prerequisite is callus induction and regeneration protocol in this indica rice genotype. The mature seeds of AC39020 used as explants for callus induction on LS, MS and N6 media with different hormones and amino acid concentrations. LS basal media with 2.5 mgL-1, 2, 4-D and 500 mgL-1 glutamine showed 91.3% callus induction frequency. Subsequently the embryogenic callus was cultured on MS media supplemented with BAP, Kinetin, NAA, and TDZ. The MS medium supplementedwith 4mg L-1 BAP and 0.5 mgL-1 NAA showed 75% regeneration efficiency. Since regeneration in indica rice varieties is tedious, far-reaching, highly genotype-specific, we exposed the embryogenic calli for mild desiccation stress for 24 h and 48 h. The desiccation treatment for 48h increased shoot regeneration frequency from 16.7 % to 40.2 % compared to non-desiccated calli. The protocol developed was highly reproducible and this protocol can also be used for further improvement of this rice genotype through genetic modification.

DOI 10.11648/j.jps.20150305.11
Published in Journal of Plant Sciences (Volume 3, Issue 5, October 2015)
Page(s) 248-254
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Callus, Regeneration, Rice, Hormone, Desiccation Stress

References
[1] Sasaki, T. (2005) The map based sequence of the rice genome. Nature, 436,793-800.
[2] Christou, P. (1997) Rice transformation: bombardment. Plant MolecularBiology, 35, 197- 203.
[3] Khush, G. S. and Virk, P. S. (2000) Rice breeding: achievements and future strategies. Crop Improvement, 27 (2), 115-144.
[4] Raina, S. K. (1989) Tissue culture in rice improvement: status and potential.Advances in Agronomy, 42, 339-398.
[5] Sikder, M. B. H., Sen, P. K., Mamun, A. A., Ali, M. R. and Rahman, S. M. (2006) In vitro regeneration of aromatic rice (Oryza sativa L.). International Journal of Agriculture Biology, 8(6), 759-760.
[6] Hoque, M. E. and Mansfield, J. W. (2004) Effect of genotype and explant age on callus induction and subsequent plant regeneration from root-derived callus of indica rice genotypes. Plant Cell Tissue Organ Culture, 78, 217-223.
[7] Rueb, S., Leneman, M., Schilperoort, R. A. and Hensgens, L. A. M. (1994) Efficient plant regeneration through somatic embryogenesis from callus induced on mature rice embryos (Oryza sativa L.). Plant Cell, Tissue and Organ Culture, 36, 259-264.
[8] Pravin, V. J., Mahendra, D., Salujat.,Sarawgi, A. K., Saxena, R. A. V. and Girish, C. (2011) Assessment of critical factors influencing callus induction, in vitro regeneration and selection of bombarded indica rice genotypes. Journal of Cell & Plant Science, 2 (2), 24-42.
[9] Chu, Q. R. and Croughan, T. P. (1990) Genetics of plant regeneration in immature panicle culture of rice. Crop Science, 30, 1194-1197.
[10] Mandal, N. and Gupta, S. (1995) Effect of and culture medium on androgenic callus formation and green plant regeneration in indica rice. Indian Journal of Experimental Biology, 33, 761-765.
[11] Zhu, Y., Ouyang, W., Li, Y. and Chen, Z. (1996) The effects of 2ip and 2, 4-D on rice calli differentiation. Plant Growth Regulators, 19, 19-24.
[12] Jain, R. K. (1997) Effect of some factors on plant regeneration from indica rice callus and protoplasts- a review. Indian Journal of Experimental Biology, 35, 323-331.
[13] Rance, I. M., Tian, W., Mathews, H., Dekochko, A., Beachy, R. N. and Fauquet, C. (1994) Partial desiccation of mature embryo-derived calli, a simple treatment that dramatically enhances the regeneration ability of indica rice. Plant Cell Reports, 13, 647-651.
[14] Biswas, A. and Mandal, A. B. (2007) Plant regeneration in different genotypes of indica rice. Indian Journal of Biotechnology, 6(4), 532-540.
[15] Wani, S. H., Sanghera, G. S. and Gosal, S. S. (2011) An efficient and reproducible method for regeneration of whole plants from mature seeds of a high yielding indica rice (Oryza sativa L.) Variety PAU 201. New Biotechnology, 28, 418-422.
[16] Hiei, Y. and Komari. T. (2008) Agrobacterium-mediated transformation of rice using immature embryos or calli induced from mature seed. Nature Protocals 3: 824-834.
[17] Joyia, F. A. and Khan, M. S. (2012) Reproducible and expedient rice regeneration system using in vitro grown plants. African Journal of Biotechnology, 11, 138-144.
[18] Yookongkaew, N., Srivatanakul, M and Narangajavana, J. (2007) Development of genotype-independent regeneration system for transformation of rice (Oryza sativa ssp. Indica). Journal of Plant Research, 120, 237-245.
[19] Puhan, P. and Siddiq, E. A. (2013) Protocol optimization and evaluation of rice varieties response to In vitro regeneration. Advances in Bioscience and Biotechnology, 4, 647-653.
[20] Yan, C. J. and Zhao, Q. H. (1982) Callus induction in plantlet regeneration from leaf blade of OryzasativaL. Subspindica. Plant Science Letter, 29. 175-182.
[21] Genovesi, A. D and Magill, C. W. (1979) Improved rate of callus and plant regeneration from rice anther culture following cold shock. Crop Science, 19, 662–664.
[22] Ramesh, M., Murugiah, V. and Gupta, A. K. (2009) Efficient in vitro plant regeneration via leaf base segments of indica rice (Oryza sativa L.). Indian Journal of Experimental Biology, 47, 68-74.
[23] Sharma, C., Kaur, M., Kaur, A. andGosal, S.S. (2012) In vitro plant regeneration studies in three indica rice varieties. International Journal of Agriculture and Environmental Biotechnology, 5(4), 309-313.
[24] Huetteman, A. and Preece, E. J. (1993) Thiodiazuran: a potent cytokinin for woody plant tissue culture. Plant Cell, Tissue and Organ Culture, 33, 105-119.
[25] Dey, M., Bakshi, S., Galiba, G., Sahoo, L. and Pand, S. K. (2012) Development of a genotype independent and transformation amenable regeneration system from shoot apex in rice (Oryza sativa spp. Indica) using TDZ. 3 Biotech, 2(3), 233-240.
[26] Rafique, M. Z., Rashid, H., Chaudhary, M. F., Chaudhry, Z. and Cheema, N. M. (2011) Study on callogenesis and organogenesis inlocal cultivars of rice (Oryza sativa l.) Pakistan Journal of Botany, 43(1), 191-203.
[27] Alam, M. J., Imran, M., Hassan, L., Rubel, M. H. and Shamsuddoha, M. J. (2012). In vitro Regeneration of High Yielding Indica Rice (Oryzasativa L.) Varieties, Journal of Environmental science & Natural Resources, 5, 173-177.
[28] Chand, S. and Sahrawat, A. K. (2000) Stimulatory effect of partial desiccation in plant regeneration in indica rice (Oryza sativa. L). Journal of plant biochemistry and biotechnology, 10(1), 43-47.
[29] Saharan, V., Yadav, R. C., Yadav, N. R. and Chapagain, B. P. (2004) High frequency plant regeneration from desiccated calli of indica rice (Oryza sativa L.). African Journal of Biotechnology, 3 (5), 256-259.
[30] Makerlya, H., Rahmata, Z. and Wagirana, A. (2012) Potential use of partial desiccation treatment for regeneration system of malaysianindica rice (Oryza sativa L.) Jurnalteknologi (sciences & engineering), 59 (1), 97-100.
[31] Wagiran, A. I. I., Zain, C. R. C. M. and Abdullah, R. (2008) Improvement of plant regeneration from embryogenic suspension cell culture of japonica rice. Journal of Biological Sciences, 8, 570-576.
[32] Murashige, T. and Skoog, F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. PhysiologiaPlantarum, 15, 473-497.
[33] Linsmaier, E. M. and Skoog, F. (1965) Organic growth factor requirements of tobacco tissue culture. Physiol. Plant, 18, 100-127.
[34] Chu, C. C., Wang, C. C., Sun, C. S., Hsu, C., Yin, K. C. and Chu, C. Y. (1975) Establishment of an efficient medium for anther culture in rice through comparative experiments on the nitrogen sources. ScientiaSinica, 18, 659-668.
[35] Islam, M. M., Ahmed, M. and Mahaldar, D. (2005) In vitro callus induction and plant regeneration in seed explants of rice (Oryza sativa L). Research Journal of Agriculture and Biological Sciences, 1(1), 72-75.
[36] Pandey SK, Ramesh B and Gupta PK (1994) Study on effect on genotype and culture medium on callus formation and plant regeneration in rice (Oryzasativa L.). Indian Journal of Genetics. 54: 293-299.
[37] Bajaj, Y. P. S., 1991. Biotechnology in Rice Improvement. In Biotechnology in Agriculture and Forestry 14. Eds. Y. P. S. Bajaj. Rice Spriger-Verlag, pp: 1-18.
[38] Libin, A.; King, P. J. H.; Ong, K. H.; Chubo, J. K. and Sipen P. (2012). Callus induction and plant regeneration of Sarawak rice (OryzasativaL.) variety Biris. African Journal of Agricultural Research, 7(30): 4260-4265.
[39] Mahajan, R., Aslam, L. and Kousar, H. (2013) Effect of growth regulators on in vitro cultures of two basmati rice genotypes: ranbir basmati and basmati 370. International Journal of Pharmaceutical, Chemical and Biological Sciences, 3(4), 1131-1138.
[40] Aditya, T. L., Hoque, M. E. and Khalequzzaman, M.(2004) Response to high frequency callus induction ability from root regions of germinated embryo in indica rice. Pakistan Journal of Biological Sciences, 7(5), 861-864.
[41] Khanna, H. K. and Raina, S. K. (1998) Genotype × culture media interaction effects on regeneration response of three indica rice cultivars. Plant Cell, Tissue and Organ Culture, 52, 145-153.
[42] Lee K, Jeon H, Kim M (2002). Optimization of a mature embryo-based in vitro culture system for high frequency somatic embryogenic callus induction and plant regeneration from japonica rice cultivars. Plant Cell, Tissue and Organ Culture 71: 237-244.
[43] Khatun, M. M., Ali, M. H. and Desamero, N. V. (2003) Effect of genotype and culture media on callus formation and plant regeneration from mature seed scutella culture in rice. Plant Tissue Culture, 13(2), 99-107.
[44] Tsukuhara, M and Hirosawa, T. (1992) Simple dehydration treatment promotes plantlet regeneration of rice (Oryzasativa L.) Callus. Plant Cell Reports, 11 (11), 550-3.
[45] Jain, R. K., Jain, S. and Wu, R. (1996) Stimulatory effect of water stress on plant regeneration in aromatic indica rice varieties. Plant cell Reports, 15(6), 449-454.
[46] Haq, I. Zhao, C. X. Mukhtar, Z. Jaleel, C. A. and Azooz, M. M. (2009) Effect of physical desiccation on plant regeneration efficiency in rice (Oryza sativa L.). Journal of Plant Physiology, 166, 1569-1575.
[47] Singh, B. (2014) Effect of desiccation and chilling treatment on somatic embryo development and germination in rough lemon (citrus jambhiri lush). British biotechnology journal 4(2): 136-148.
[48] Wenzhong, T., Rance, I., Sivamani, E., Fauquet, C. and Beachy, R. N. (1994) Improvement of plant regeneration frequency in vitro in indica rice. Chinese Journal of Genetics, 21, 1-9.
[49] Yang, Y. S., Y. D. Zheng, Y. L. Chen and Y. Y. Jian, 1999. Improvement of plant regeneration from long-term cultured calluses of Taipei 309, a model rice variety in in vitro studies. Plant Cell, Tissue and Organ Culture. 57: 199-206.
Author Information
  • Department of Botany, S.K. University, Anantapur, Andhra Pradesh, India; Department of Crop Physiology, UAS, GKVK, Bangalore, Karnataka, India

  • Department of Crop Physiology, UAS, GKVK, Bangalore, Karnataka, India; Plant Biology Division, the Samuel Roberts Noble Foundation, Ardmore, USA

  • Department of Crop Physiology, UAS, GKVK, Bangalore, Karnataka, India

  • Department of Crop Physiology, UAS, GKVK, Bangalore, Karnataka, India

  • Department of Botany, S.K. University, Anantapur, Andhra Pradesh, India

  • Department of Botany, S.K. University, Anantapur, Andhra Pradesh, India

  • Department of Botany, S.K. University, Anantapur, Andhra Pradesh, India

Cite This Article
  • APA Style

    Amaranatha Reddy Vennapusa, Ramu S. Vemanna, Rajashekar Reddy B. H., K. C. Babitha, K. Kiranmai, et al. (2015). An Efficient Callus Induction and Regeneration Protocol for a Drought Tolerant Rice Indica Genotype AC39020. Journal of Plant Sciences, 3(5), 248-254. https://doi.org/10.11648/j.jps.20150305.11

    Copy | Download

    ACS Style

    Amaranatha Reddy Vennapusa; Ramu S. Vemanna; Rajashekar Reddy B. H.; K. C. Babitha; K. Kiranmai, et al. An Efficient Callus Induction and Regeneration Protocol for a Drought Tolerant Rice Indica Genotype AC39020. J. Plant Sci. 2015, 3(5), 248-254. doi: 10.11648/j.jps.20150305.11

    Copy | Download

    AMA Style

    Amaranatha Reddy Vennapusa, Ramu S. Vemanna, Rajashekar Reddy B. H., K. C. Babitha, K. Kiranmai, et al. An Efficient Callus Induction and Regeneration Protocol for a Drought Tolerant Rice Indica Genotype AC39020. J Plant Sci. 2015;3(5):248-254. doi: 10.11648/j.jps.20150305.11

    Copy | Download

  • @article{10.11648/j.jps.20150305.11,
      author = {Amaranatha Reddy Vennapusa and Ramu S. Vemanna and Rajashekar Reddy B. H. and K. C. Babitha and K. Kiranmai and A. Nareshkumar and Chinta Sudhakar},
      title = {An Efficient Callus Induction and Regeneration Protocol for a Drought Tolerant Rice Indica Genotype AC39020},
      journal = {Journal of Plant Sciences},
      volume = {3},
      number = {5},
      pages = {248-254},
      doi = {10.11648/j.jps.20150305.11},
      url = {https://doi.org/10.11648/j.jps.20150305.11},
      eprint = {https://download.sciencepg.com/pdf/10.11648.j.jps.20150305.11},
      abstract = {Most of the cultivated indica rice genotypes are less amenable to genetic modifications due to their poor callus induction and regeneration potential. The prerequisite for genetic enhancement of indica rice genotypes by biotechnological approach is to develop an efficient protocol for callus induction and plant regeneration. In the present study, we established an optimized regeneration protocol for rice genotype AC39020, which is moderately drought tolerant with high root growth and biomass. To use this genotype in the crop improvement program the prerequisite is callus induction and regeneration protocol in this indica rice genotype. The mature seeds of AC39020 used as explants for callus induction on LS, MS and N6 media with different hormones and amino acid concentrations. LS basal media with 2.5 mgL-1, 2, 4-D and 500 mgL-1 glutamine showed 91.3% callus induction frequency. Subsequently the embryogenic callus was cultured on MS media supplemented with BAP, Kinetin, NAA, and TDZ. The MS medium supplementedwith 4mg L-1 BAP and 0.5 mgL-1 NAA showed 75% regeneration efficiency. Since regeneration in indica rice varieties is tedious, far-reaching, highly genotype-specific, we exposed the embryogenic calli for mild desiccation stress for 24 h and 48 h. The desiccation treatment for 48h increased shoot regeneration frequency from 16.7 % to 40.2 % compared to non-desiccated calli. The protocol developed was highly reproducible and this protocol can also be used for further improvement of this rice genotype through genetic modification.},
     year = {2015}
    }
    

    Copy | Download

  • TY  - JOUR
    T1  - An Efficient Callus Induction and Regeneration Protocol for a Drought Tolerant Rice Indica Genotype AC39020
    AU  - Amaranatha Reddy Vennapusa
    AU  - Ramu S. Vemanna
    AU  - Rajashekar Reddy B. H.
    AU  - K. C. Babitha
    AU  - K. Kiranmai
    AU  - A. Nareshkumar
    AU  - Chinta Sudhakar
    Y1  - 2015/09/07
    PY  - 2015
    N1  - https://doi.org/10.11648/j.jps.20150305.11
    DO  - 10.11648/j.jps.20150305.11
    T2  - Journal of Plant Sciences
    JF  - Journal of Plant Sciences
    JO  - Journal of Plant Sciences
    SP  - 248
    EP  - 254
    PB  - Science Publishing Group
    SN  - 2331-0731
    UR  - https://doi.org/10.11648/j.jps.20150305.11
    AB  - Most of the cultivated indica rice genotypes are less amenable to genetic modifications due to their poor callus induction and regeneration potential. The prerequisite for genetic enhancement of indica rice genotypes by biotechnological approach is to develop an efficient protocol for callus induction and plant regeneration. In the present study, we established an optimized regeneration protocol for rice genotype AC39020, which is moderately drought tolerant with high root growth and biomass. To use this genotype in the crop improvement program the prerequisite is callus induction and regeneration protocol in this indica rice genotype. The mature seeds of AC39020 used as explants for callus induction on LS, MS and N6 media with different hormones and amino acid concentrations. LS basal media with 2.5 mgL-1, 2, 4-D and 500 mgL-1 glutamine showed 91.3% callus induction frequency. Subsequently the embryogenic callus was cultured on MS media supplemented with BAP, Kinetin, NAA, and TDZ. The MS medium supplementedwith 4mg L-1 BAP and 0.5 mgL-1 NAA showed 75% regeneration efficiency. Since regeneration in indica rice varieties is tedious, far-reaching, highly genotype-specific, we exposed the embryogenic calli for mild desiccation stress for 24 h and 48 h. The desiccation treatment for 48h increased shoot regeneration frequency from 16.7 % to 40.2 % compared to non-desiccated calli. The protocol developed was highly reproducible and this protocol can also be used for further improvement of this rice genotype through genetic modification.
    VL  - 3
    IS  - 5
    ER  - 

    Copy | Download

  • Sections