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Research Article

volume 42 issue 2 (april 2019) : 178-181,   Doi: 10.18805/LR-3844

An efficient in vitro shoot regeneration protocol from embryo explants of chickpea (Cicer arietinum L.)

T
T. Raghavendra
V
V. Jayalakshmi
Y
Y. Padmalatha
1Department of Crop Physiology, Acharya N.G. Ranga Agricultural University, Nandyal-518 501, Andhra Pradesh, India.

  • Submitted03-02-2017|

  • Accepted17-05-2018|

  • First Online 20-08-2018|

  • doi 10.18805/LR-3844

Cite article:- Raghavendra T., Jayalakshmi V., Padmalatha Y. (2018). An efficient in vitro shoot regeneration protocol from embryo explants of chickpea (Cicer arietinum L.). Legume Research. 42(2): 178-181. doi: 10.18805/LR-3844.

ABSTRACT

Chickpea (Cicer arietinum L.) is an important grain legume in the world. Development of reliable tissue culture protocols is very important for genetic transformation studies. Direct shoot bud differentiation and multiple shoot induction from embryonal axes explants of chickpea varieties with combination of growth regulators in NBeG-3 and NBeG-72 was tried at Regional Agricultural Research Station, Nandyal in 2015-16. Explants were cultured on Murashige and Skoog (MS) medium augmented with different concentrations of Benzyl amino purine (BAP), Napthalene acetic acid (NAA), Kinetin and Indole acetic acid (IAA). The frequency of shoot bud regeneration was influenced by the preconditioning of explants with 6mg/l BAP and 0.5mg/l thidiazuron (TDZ) for 10 days resulted in enhanced frequency of multiple shoots. Among the various concentrations tested, 4.0 mg/l BAP and 2.0 mg/l Napthalene acetic acid (NAA) were found to be the best for maximum shoot bud differentiation. Elongation of multiple shoots were obtained in MS medium with the concentration 0.5 mg/l gibberillic acid (GA3). Among the tested varieties there was a variation in regeneration ability with fastest shoot bud initiation (18.5 days), and highest number of shoots per explant (5.3) and highest shoot length (3.5 cm) in NBeG 3.  

REFERENCES

  1. Aasim, M., Khawar, K.M and Ozcan, S. 2009 a. In vitro micropropagation from plumular apices of Turkish cowpea (Vignaun guiculata L.) cultivar Akkiz. Scientia. Horticulturae, 122:468-471.
  2. Aasim, M., Khawar, K.M and Özcan, S. 2010.Efficient in vitro propagation from preconditioned embryonic axes of Turkish cowpea (Vignaun guiculata L.) Cultivar akkiz. Archives of Biological Sciences, 62:1047-1052.
  3. Aasim, M., Day, S., Rezaei, F., Hajyzadeh, M., Tahir, S and Ozcan, S. 2011. In vitro shoot regeneration from preconditioned explants of chickpea (Cicer arietinum L.) cv.Gokce. African Journal of Biotechnolgy, 10(11):2020-2023.
  4. Aasim, M., Day, S., Rezaei, F and Hajyzadeh, M. 2013. Multiple shoot regeneration of plumular apices of chickpea. Turkish Journal of Agriculture and Forestry, 37:33-39.
  5. Barik, D.P., Naik, S.K.U., Nudgal, A and Chand, P.K. 2004. Rapid plant regeneration through in vitro axillary shoots proliferation of butterfly pea (Clitoria ternatea L.). In vitro Cellular and Developmental Biology-Plant, 43:144-148.
  6. Chakraborti, D., Sarkar, A and Das, S. 2006. Efficient and rapid in vitro plant regeneration system for Indian cultivars of chickpea (Cicer arietinum L.) Plant cell tissue organ culture, 86: 117-123.
  7. Jayanand, B., Sudarsanam, G and Sharma, K K. 2003. An efficient protocol for the regeneration of whole plants of chickpea (Cicer arietinum L.) by using axillary meristem explants derived from in vitro germinated seedlings. In vitro Cellular and Developmental Biology-Plant, 39: 171-179.
  8. Kiran Ghanti, S., Sujata, K G and Srinath Rao M. 2009. The effect of phenyl acetic acid on shoot bud induction, elongation and rooting of chickpea. Biologia Plantarum, 53: 779. 
  9. Malik, K.A and Saxena, P.K. 1992. Thidiazuron induces high frequency shoots regeneration in intact seedlings of pea (Pisum sativum),    Chickpea (Cicer arietinum L) and lentil (Lensculinaris). Plant Physiology, 19:731-740.
  10. Odutayo, O.I., Akinirinusi, F.B., Odunbososye, I and Oso, R.T. 2005. Multiple shoot induction from embryo derived callus cultures of cowpea (Vigna ungimculata L.). Walp. African Journal of Biotechnolgy,4:1214- 1216.
  11. Polisetty, R., Paul, V., Deveshwar, J.J., Khetarpal, S., Suresh, K and Chandra, R. 1997. Multiple shoot induction by benzyladenine and complete plant regeneration from seed explants of chickpea (Cicer arietinum L.). Plant Cell Reports, 16: 565-571.
  12. Parveen, S.,Venkateshwarlu, M., Srinivas, D., Jagan, K and Ugandhar, T. 2012. Direct in vitro shoots proliferation of chick pea (Cicer arietinum L.) from shoot tip explants induced by thidiazuron. Bioscience Discovery, 3(1): 01-05.
  13. Rizvi, S.M.H and Singh, R.P (2000). In vitro plant regeneration from immature leaflet-derived callus cultures of Cicer arietinum L. via organogenesis. Plant Cell Biotechnology and Molecular Biology, 1: 109-114.
  14. Shri, P.V and Davis T.M. 1992. Zeatin induced shoot regeneration from immature chickpea (Cicer arietinum L.) cotyledons. Plant cell tissue and organ Culture, 28- 45.
  15. Sharma, K.K and Ortiz, R. 2000. Program for the application of genetic transformation for crop improvement in the semi-arid tropics. In vitro Cellular and Developmental Biology-Plant, 36: 83-92.
  16. Sanyal, I., Singh, A.K, Kaushik, M and Amla, D.V. 2005. Agrobacterium mediated transformation of chickpea (Cicer arietinum L.) with Bacillus thuringiensis cry1Ac gene for resistance against pod borer insect Helicoverpa armigera. Plant Science, 168: 1135-1146.
  17. Tang, X.M and Chen, L. 2011. A review on regeneration in cowpea (Vigna unguiculate L. Walp). Journal of Agriculture Science and Technology. 5:4. ISSN 1939-1250, USA.
  18. Wang H.M., Liu M.H., Wang M.H and Zu, Y.G. 2008. Effects of Thidiazuron, basal medium and light quality on adventitious shoot regeneration from in vitro culture stem of Populus alba x P. berolinensis. Journal of Forestry Research, 19(3):257-259. 
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    Research Article

    volume 42 issue 2 (april 2019) : 178-181,   Doi: 10.18805/LR-3844

    An efficient in vitro shoot regeneration protocol from embryo explants of chickpea (Cicer arietinum L.)

    T
    T. Raghavendra
    V
    V. Jayalakshmi
    Y
    Y. Padmalatha
    1Department of Crop Physiology, Acharya N.G. Ranga Agricultural University, Nandyal-518 501, Andhra Pradesh, India.

    • Submitted03-02-2017|

    • Accepted17-05-2018|

    • First Online 20-08-2018|

    • doi 10.18805/LR-3844

    Cite article:- Raghavendra T., Jayalakshmi V., Padmalatha Y. (2018). An efficient in vitro shoot regeneration protocol from embryo explants of chickpea (Cicer arietinum L.). Legume Research. 42(2): 178-181. doi: 10.18805/LR-3844.

    ABSTRACT

    Chickpea (Cicer arietinum L.) is an important grain legume in the world. Development of reliable tissue culture protocols is very important for genetic transformation studies. Direct shoot bud differentiation and multiple shoot induction from embryonal axes explants of chickpea varieties with combination of growth regulators in NBeG-3 and NBeG-72 was tried at Regional Agricultural Research Station, Nandyal in 2015-16. Explants were cultured on Murashige and Skoog (MS) medium augmented with different concentrations of Benzyl amino purine (BAP), Napthalene acetic acid (NAA), Kinetin and Indole acetic acid (IAA). The frequency of shoot bud regeneration was influenced by the preconditioning of explants with 6mg/l BAP and 0.5mg/l thidiazuron (TDZ) for 10 days resulted in enhanced frequency of multiple shoots. Among the various concentrations tested, 4.0 mg/l BAP and 2.0 mg/l Napthalene acetic acid (NAA) were found to be the best for maximum shoot bud differentiation. Elongation of multiple shoots were obtained in MS medium with the concentration 0.5 mg/l gibberillic acid (GA3). Among the tested varieties there was a variation in regeneration ability with fastest shoot bud initiation (18.5 days), and highest number of shoots per explant (5.3) and highest shoot length (3.5 cm) in NBeG 3.  

    REFERENCES

    1. Aasim, M., Khawar, K.M and Ozcan, S. 2009 a. In vitro micropropagation from plumular apices of Turkish cowpea (Vignaun guiculata L.) cultivar Akkiz. Scientia. Horticulturae, 122:468-471.
    2. Aasim, M., Khawar, K.M and Özcan, S. 2010.Efficient in vitro propagation from preconditioned embryonic axes of Turkish cowpea (Vignaun guiculata L.) Cultivar akkiz. Archives of Biological Sciences, 62:1047-1052.
    3. Aasim, M., Day, S., Rezaei, F., Hajyzadeh, M., Tahir, S and Ozcan, S. 2011. In vitro shoot regeneration from preconditioned explants of chickpea (Cicer arietinum L.) cv.Gokce. African Journal of Biotechnolgy, 10(11):2020-2023.
    4. Aasim, M., Day, S., Rezaei, F and Hajyzadeh, M. 2013. Multiple shoot regeneration of plumular apices of chickpea. Turkish Journal of Agriculture and Forestry, 37:33-39.
    5. Barik, D.P., Naik, S.K.U., Nudgal, A and Chand, P.K. 2004. Rapid plant regeneration through in vitro axillary shoots proliferation of butterfly pea (Clitoria ternatea L.). In vitro Cellular and Developmental Biology-Plant, 43:144-148.
    6. Chakraborti, D., Sarkar, A and Das, S. 2006. Efficient and rapid in vitro plant regeneration system for Indian cultivars of chickpea (Cicer arietinum L.) Plant cell tissue organ culture, 86: 117-123.
    7. Jayanand, B., Sudarsanam, G and Sharma, K K. 2003. An efficient protocol for the regeneration of whole plants of chickpea (Cicer arietinum L.) by using axillary meristem explants derived from in vitro germinated seedlings. In vitro Cellular and Developmental Biology-Plant, 39: 171-179.
    8. Kiran Ghanti, S., Sujata, K G and Srinath Rao M. 2009. The effect of phenyl acetic acid on shoot bud induction, elongation and rooting of chickpea. Biologia Plantarum, 53: 779. 
    9. Malik, K.A and Saxena, P.K. 1992. Thidiazuron induces high frequency shoots regeneration in intact seedlings of pea (Pisum sativum),    Chickpea (Cicer arietinum L) and lentil (Lensculinaris). Plant Physiology, 19:731-740.
    10. Odutayo, O.I., Akinirinusi, F.B., Odunbososye, I and Oso, R.T. 2005. Multiple shoot induction from embryo derived callus cultures of cowpea (Vigna ungimculata L.). Walp. African Journal of Biotechnolgy,4:1214- 1216.
    11. Polisetty, R., Paul, V., Deveshwar, J.J., Khetarpal, S., Suresh, K and Chandra, R. 1997. Multiple shoot induction by benzyladenine and complete plant regeneration from seed explants of chickpea (Cicer arietinum L.). Plant Cell Reports, 16: 565-571.
    12. Parveen, S.,Venkateshwarlu, M., Srinivas, D., Jagan, K and Ugandhar, T. 2012. Direct in vitro shoots proliferation of chick pea (Cicer arietinum L.) from shoot tip explants induced by thidiazuron. Bioscience Discovery, 3(1): 01-05.
    13. Rizvi, S.M.H and Singh, R.P (2000). In vitro plant regeneration from immature leaflet-derived callus cultures of Cicer arietinum L. via organogenesis. Plant Cell Biotechnology and Molecular Biology, 1: 109-114.
    14. Shri, P.V and Davis T.M. 1992. Zeatin induced shoot regeneration from immature chickpea (Cicer arietinum L.) cotyledons. Plant cell tissue and organ Culture, 28- 45.
    15. Sharma, K.K and Ortiz, R. 2000. Program for the application of genetic transformation for crop improvement in the semi-arid tropics. In vitro Cellular and Developmental Biology-Plant, 36: 83-92.
    16. Sanyal, I., Singh, A.K, Kaushik, M and Amla, D.V. 2005. Agrobacterium mediated transformation of chickpea (Cicer arietinum L.) with Bacillus thuringiensis cry1Ac gene for resistance against pod borer insect Helicoverpa armigera. Plant Science, 168: 1135-1146.
    17. Tang, X.M and Chen, L. 2011. A review on regeneration in cowpea (Vigna unguiculate L. Walp). Journal of Agriculture Science and Technology. 5:4. ISSN 1939-1250, USA.
    18. Wang H.M., Liu M.H., Wang M.H and Zu, Y.G. 2008. Effects of Thidiazuron, basal medium and light quality on adventitious shoot regeneration from in vitro culture stem of Populus alba x P. berolinensis. Journal of Forestry Research, 19(3):257-259. 
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