In vitro plant regeneration in rough lemon (Citrus jambhiri Lush.) through epicotyl segments by direct shoot organogenesis
Article Main
Abstract
The effect of Murashige and Skoog (MS) medium supplemented with various concentrations and combinations of growth hormones on direct regeneration from one month old epicotyl segments of in vitro grown rough lemon (Citrus jambhiri Lush.) seedlings was studied. The earliest bud induction in 7.5 days, highest bud induction frequency (98.50%), percent regeneration(90.53) were obtained on MS medium supplemented with 6-Benzylaminopurine (BAP) (1mglit-1) with an average number of 12.50 buds per explants. The epicotyls segments with proliferated buds were transferred to elongation media in order to improve the recovery of normal shoots. Maximum number of elongated shoots (8.50) was obtained on MS medium having BAP (0.5mglit-1) + Gibberellic Acid (GA3)(1.0 mglit-1).These elongated shoots were then rooted on MS medium containing Indole-3-butyric acid (IBA) (0.1mglit-1) + Indole-3-aceticacid(IAA)(0.5mglit-1) with highest rooting percentage(96%) and root number(5.0). Early (10.10 days) rooting was observed in MS medium supplemented with NAA1.0 mglit-1 + IBA0.5 mglit-1.The plantlet survival was 98.52%, when plantlets were transferred to plastic pots containing a mixture of garden soil and vermiculite (1:1). The hardened plants were successfully established in the soil. The present study developed protocol which can be reliably used for in vitro regeneration of rough lemon and for gene transfer studies in rough lemon, especially to induce salinity and Phytophthora tolerance.
Article Details
Article Details
Keywords
Citrus jambhiri Lush., Direct regeneration, Epicotyl segments, In vitro, Rooting, Rough lemon
References
Ali, S. and Mirza, B. (2006). Micropropagation of rough lemon(Citrus jambhiri Lush.):Effect of explant type and hormone concentration. Acta Botanica Croatica.65(2):137-146.
Almeida, W.A.B., Filho, F.A.A.M., Mendes, B.M.J. and Rodri-guez, A.P.M. (2002). In vitro organogenesis optimiza-tion and plantlet regeneration in Citrus sinensis and Citrus limonia. Scientia Agricola. 59(1): 35-40.
Altaf, N. and Ahmad, M.S. (1997). Effect of culture medium composition on nucellar embryogenesis in three citrus cultivars: Kinnow, Jatti Khatti (Rough lemon) and Gada Dehi. In Endeavors in Biotechnology. Pp. 25-30.Edited by Ihsan Iiahi, Department of Botany, University of Peshawar.
Bordon, Y., Guardiola, J.I. and Garcia-Luis, A. (2000). Genotype affects the morphogenic response invitro epicotyl segments of citrus rootstocks. Annals of Bot-any. 86:156-166.
Cervera, M., Ortega, C., Navarro, A., Navarro, L. and Pena , L. (2000). Generation of Transgenic citrus plants with tolerance to salinity gene HAL2 from yeast. The Jour-nal of Horticultural Science and Biotechnology. 75(1):26–30.
Chakravarty, B. and Goswami, B.C. (1999). Plantlet regeneration from term callus cultures of Citrus acida Roxb. and the uniformity of regenerated plants. Scientia Horticultura. 82(1-2):159-69.
Chaturvedi, H.C., Singh, S.K., Sharma, A.K. and Agnihotri, S.(2001). Citrus tissue culture employing vegetative explants. Indian Journal of Experimental Biology. 39(11): 1080–1095.
Costa, M.G.C., Otoni, W.C. and Moor, G.A. (2002). A ele-vation of factors affecting the efficiency of Agrobacterium mediated transformation of Citrus paradise Macf. and production of transgenic plants containing carotenoid biosynthetic genes. Plant Cell Reports. 21(4): 365-373.
Das, A., Paul, A.K. and Chaudhuri, S. (2000). Micropropaga-tion of sweet orange (Citrus sinensis Osb.) for the development of nucellar seedlings. Indian Journal of Experimental Biology. 38: 269-272.
Dejam, M., Khosh-Khui, M. and Shekafandeh, A. (2006). Adventitious bud induction and plant regeneration in epicotyl segments of Bakrai (Citrus reticulata Blanco. x Citrus limetta Swing.). International Journal of Agricultural Research.1(1):14-19.
Dominguez, A., Guerri, J., Cambra, M., Navarro, L., Moreno, P., and Pena, L.S. (2000). Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus. Plant Cell Reports .19(4): 427-433.
Garcia-Luis, A., Bordon, Y., Moreira-Dias, J.M., Molina, R. and Guardiola, J.L. (1999). Explant orientation and polarity determine the morphogenic response of epicotyl segments of Troyer citrange. Annals of Botany(London) .84: 715–723.
Ghorbel, R., Juarez, J., Navarro, L., Pena, L. (1999). Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants. Theoretical and Applied Genetics. 99(1-2): 350–358.
Gill, M.I.S. and Gosal S.S. (2002). Micropropagation of pectinifera (Citrus depressa Hayata) a potential citrus rootstock for sweet orange. Indian Journal of Citriculture. 1(1):32-37.
Gmitter, F.G., Grosser, J.W. and Moore, G.A. (1992). Citrus. In. F.A. Hamerschlag and R.E. Litz (Editors), Biotechnology of Perennial Fruit Crops. CAB International, Walling-ford, pp:335-369.
Grosser, J.W. and JR. Gmitter, F.G. (2005). Applications of somatic hybridization and cybridization in crop improvement, with citrus as a model. In vitro Cellular and Developmental Biology.41:220-225.
Guo, W.W. and Deng, X.X .(2001). Wide somatic hybrids of citrus with its related genera and their potential in ge-netic improvement. Euphytica .118:175-183.
Hidaka, T. and Omura, M. (1989). Control of embryogenesis in citrus cell culture regeneration protoplasts and attempts to callus culture bank. Bulletin of the fruit tree research station, series okitsu. 16:1-17.
Hiregoudar, L., Ashok Kumar, H.G. and Murthy, M.N.(2005). In vitro culture of Feronia lemonia (L.) Swingle from hypocotyl and internodal explants. Biologia Plantarum. 49(1):41-45.
Kaneyoshi, J., Kobayashi, S., Nakamura, Y., Shigemoto, N., and Doi, Y. (1994). A simple and efficient gene transfer system to trifoliate orange (Poincirus trifoliate Raf). Plant Cell Reports. 13(10): 541- 545.
Lesham, Y. (1973). Molecular and hormonal basis of plant growth regulation( Pergamon Press,New York), 35-86.
Moreira-Dias, J.M., Molina, R.V., Bordon, Y., Guardiola, J.L. and Garcia-Luis, A .(2000). Direct and indirect shoot organogenic pathways in epicotyl cuttings of Troyer citrange differ in hormone requirements and in their response to light. Annals of Botany(London).85(1):103-110.
Moreira-Dias, J,M,, Molina, R,V., Guardiola, J.L. and Garcia-Luis, A. (2001). Day-length and photon flux density influence the growth regulator effects on morphogenesis in epicotyl segments of Troyer citrange. Scientia Horticulturae.87(4): 275-290.
Moura, T.L.M., Almeida, W.A.B., Mendes, B.M.J. and Mourao Filho, F.A.A . (2001). Organogênese in vitro de Citrus em função de concentrações de BAP e secciona-mento do explante. Revista Brasileira de Fruticultura. 23(2): 240-245.
Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiologia Plantarum. 15:473-479.
Murashige, T. And Tucker, D.P.H. (1969). Growth factor requirement of citrus tissue culture. In: International Citrus Symposium,1, Riverside. Proceedings. Riverside: University of California. 3:1155-1169.
Nwe, Y.Y., Myint, K.T., Mochizuki, Y., Vazirzanjani, M., Okayasu, K., Suzuki, S. and Ogiwara, I. (2014). In vitro regeneration through direct shoot organogenesis in Honey Orange (Citrus tangerina). Plant Biotechnology. 31(4): 341–344.
Oh, S.D., Song, W.S., Cho, H.M. and Baek, S.H. (1991). Plant regeneration in dangyooza (Citrus grandis Osb.) through somatic embryogenesis. 3. Effect of different combinations or alone of complex addenda, sucrose and 2,4-D on somatic embryogenesis. Research Reports of the Rural Development Administration Biotechnology. 33: 33-37.
Omura, M and Hidaka, T. (1992). Shoot tip culture of citrus. Bulletin of the Fruit Tree Research Station. 22 : 23-35.
Parkash, O. (2003). Studies on invitro plantlet production and acclimatization of rough lemon (Citrus jambhiri Lush.). M.Sc. Thesis, Punjab Agricultural University, Ludhiana.
Pena, L. and Navarro, L. (1999). Transgenic citrus. In: Y.P.S.(ed.), Biotechnology in Agriculture and Forestry, Vol.44,Transgenic Tress, pp.39-54. Springer-Verlag, Berlin, Germany.
Raman, H., Gosal, S.S. and Brar, D.S. (1992). Plant regen-eration from callus culture of Citrus limon and C jamb-hiri. Crop Improvement.19 (2):100-103.
Rashad, M., Khan, M.M., Ramzan, R., Adnan, S. and Khan F.A. (2005). In vitro regeneration and somatic embryo-genesis in Citrus autantifolia and Citrus sinensis. International Journal of Agriculture and Biology. 7(2): 518-520.
Rattanpal, H.S., Kaur, G. and Gupta, M. (2011). In vitro plant regeneration in rough lemon (Citrus jambhiri Lush) by direct organogenesis. African Journal of Bio-technology, 10(63):13724-13728.
Rezadost, M.H., Sohani, M.M., Hatamzadeh, A. and Reza, M. (2013). In vitro regeneration of sour orange (Citrus aurantium L.) via direct organogenesis. Plant Knowledge Journal. 2(4):150-156 .
Roistacher, C.N., Navarro, L. and Murashige, T. (1976). Recovery of citrus selections free of several viruses, exocortis viroid and spiroplasma citri by shoot-tip graft-ing in vitro. Proceedings 7th Conference International Organization Citrus Virologists, IOCV, Riverside, 186–193.
Saini, H.K., Gill, M.S. and Gill, M.I.S. (2010). Direct shoot organogenesis and plant regeneration in rough lemon (Citrus jambhiri Lush.). Indian Journal of Biotechnol-ogy. 9(4): 419-423.
Silva, R.P., Costa, M.A.P.C., Souza, A.S. and Almeida, W.A.B. (2005). Regeneracao de plantas de laranja-Pera via organogênese invitro. Pesquisa Agropecuaria Bra-sileira. 40(12):1153-1159.
Silva, R.P., Souza, A.J., Mendes, B.M.J. and Mourao Filho, F.A.A. (2010). Sour orange bud regeneration and in vitro plant development related to culture medium com-position and explant type. Revista Brasileira de Fruticultura .32(1):1-8.
Singh, S., Bansal, M.L., Singh, T.P. and Kumar, P. (1998). Statistical Methods for Research Workers. Kalyani Publishers, New Delhi.
Te-Chato, S. and Nudoung, S. (1998). Tissue culture of Citrus reticultata Blanco cv. Shogum and gene transfor-mation by Agrobacteria. Proc. Abstracts IMT-GT UNITE Conf. Songkxhla.
Thorpe, T.D. and Harry, I.S. (1997). Application of tissue culture to horticulture (In: Horticultural biotechnology, edited by A Altaian & M Ziv. Acta Horticulturae. 447:39-49.
Torres, K.C. (1988). Tissue culture technique for horticultural crops (Vam Mostrand Reinhold, New York), 295-298.
Almeida, W.A.B., Filho, F.A.A.M., Mendes, B.M.J. and Rodri-guez, A.P.M. (2002). In vitro organogenesis optimiza-tion and plantlet regeneration in Citrus sinensis and Citrus limonia. Scientia Agricola. 59(1): 35-40.
Altaf, N. and Ahmad, M.S. (1997). Effect of culture medium composition on nucellar embryogenesis in three citrus cultivars: Kinnow, Jatti Khatti (Rough lemon) and Gada Dehi. In Endeavors in Biotechnology. Pp. 25-30.Edited by Ihsan Iiahi, Department of Botany, University of Peshawar.
Bordon, Y., Guardiola, J.I. and Garcia-Luis, A. (2000). Genotype affects the morphogenic response invitro epicotyl segments of citrus rootstocks. Annals of Bot-any. 86:156-166.
Cervera, M., Ortega, C., Navarro, A., Navarro, L. and Pena , L. (2000). Generation of Transgenic citrus plants with tolerance to salinity gene HAL2 from yeast. The Jour-nal of Horticultural Science and Biotechnology. 75(1):26–30.
Chakravarty, B. and Goswami, B.C. (1999). Plantlet regeneration from term callus cultures of Citrus acida Roxb. and the uniformity of regenerated plants. Scientia Horticultura. 82(1-2):159-69.
Chaturvedi, H.C., Singh, S.K., Sharma, A.K. and Agnihotri, S.(2001). Citrus tissue culture employing vegetative explants. Indian Journal of Experimental Biology. 39(11): 1080–1095.
Costa, M.G.C., Otoni, W.C. and Moor, G.A. (2002). A ele-vation of factors affecting the efficiency of Agrobacterium mediated transformation of Citrus paradise Macf. and production of transgenic plants containing carotenoid biosynthetic genes. Plant Cell Reports. 21(4): 365-373.
Das, A., Paul, A.K. and Chaudhuri, S. (2000). Micropropaga-tion of sweet orange (Citrus sinensis Osb.) for the development of nucellar seedlings. Indian Journal of Experimental Biology. 38: 269-272.
Dejam, M., Khosh-Khui, M. and Shekafandeh, A. (2006). Adventitious bud induction and plant regeneration in epicotyl segments of Bakrai (Citrus reticulata Blanco. x Citrus limetta Swing.). International Journal of Agricultural Research.1(1):14-19.
Dominguez, A., Guerri, J., Cambra, M., Navarro, L., Moreno, P., and Pena, L.S. (2000). Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus. Plant Cell Reports .19(4): 427-433.
Garcia-Luis, A., Bordon, Y., Moreira-Dias, J.M., Molina, R. and Guardiola, J.L. (1999). Explant orientation and polarity determine the morphogenic response of epicotyl segments of Troyer citrange. Annals of Botany(London) .84: 715–723.
Ghorbel, R., Juarez, J., Navarro, L., Pena, L. (1999). Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants. Theoretical and Applied Genetics. 99(1-2): 350–358.
Gill, M.I.S. and Gosal S.S. (2002). Micropropagation of pectinifera (Citrus depressa Hayata) a potential citrus rootstock for sweet orange. Indian Journal of Citriculture. 1(1):32-37.
Gmitter, F.G., Grosser, J.W. and Moore, G.A. (1992). Citrus. In. F.A. Hamerschlag and R.E. Litz (Editors), Biotechnology of Perennial Fruit Crops. CAB International, Walling-ford, pp:335-369.
Grosser, J.W. and JR. Gmitter, F.G. (2005). Applications of somatic hybridization and cybridization in crop improvement, with citrus as a model. In vitro Cellular and Developmental Biology.41:220-225.
Guo, W.W. and Deng, X.X .(2001). Wide somatic hybrids of citrus with its related genera and their potential in ge-netic improvement. Euphytica .118:175-183.
Hidaka, T. and Omura, M. (1989). Control of embryogenesis in citrus cell culture regeneration protoplasts and attempts to callus culture bank. Bulletin of the fruit tree research station, series okitsu. 16:1-17.
Hiregoudar, L., Ashok Kumar, H.G. and Murthy, M.N.(2005). In vitro culture of Feronia lemonia (L.) Swingle from hypocotyl and internodal explants. Biologia Plantarum. 49(1):41-45.
Kaneyoshi, J., Kobayashi, S., Nakamura, Y., Shigemoto, N., and Doi, Y. (1994). A simple and efficient gene transfer system to trifoliate orange (Poincirus trifoliate Raf). Plant Cell Reports. 13(10): 541- 545.
Lesham, Y. (1973). Molecular and hormonal basis of plant growth regulation( Pergamon Press,New York), 35-86.
Moreira-Dias, J.M., Molina, R.V., Bordon, Y., Guardiola, J.L. and Garcia-Luis, A .(2000). Direct and indirect shoot organogenic pathways in epicotyl cuttings of Troyer citrange differ in hormone requirements and in their response to light. Annals of Botany(London).85(1):103-110.
Moreira-Dias, J,M,, Molina, R,V., Guardiola, J.L. and Garcia-Luis, A. (2001). Day-length and photon flux density influence the growth regulator effects on morphogenesis in epicotyl segments of Troyer citrange. Scientia Horticulturae.87(4): 275-290.
Moura, T.L.M., Almeida, W.A.B., Mendes, B.M.J. and Mourao Filho, F.A.A . (2001). Organogênese in vitro de Citrus em função de concentrações de BAP e secciona-mento do explante. Revista Brasileira de Fruticultura. 23(2): 240-245.
Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiologia Plantarum. 15:473-479.
Murashige, T. And Tucker, D.P.H. (1969). Growth factor requirement of citrus tissue culture. In: International Citrus Symposium,1, Riverside. Proceedings. Riverside: University of California. 3:1155-1169.
Nwe, Y.Y., Myint, K.T., Mochizuki, Y., Vazirzanjani, M., Okayasu, K., Suzuki, S. and Ogiwara, I. (2014). In vitro regeneration through direct shoot organogenesis in Honey Orange (Citrus tangerina). Plant Biotechnology. 31(4): 341–344.
Oh, S.D., Song, W.S., Cho, H.M. and Baek, S.H. (1991). Plant regeneration in dangyooza (Citrus grandis Osb.) through somatic embryogenesis. 3. Effect of different combinations or alone of complex addenda, sucrose and 2,4-D on somatic embryogenesis. Research Reports of the Rural Development Administration Biotechnology. 33: 33-37.
Omura, M and Hidaka, T. (1992). Shoot tip culture of citrus. Bulletin of the Fruit Tree Research Station. 22 : 23-35.
Parkash, O. (2003). Studies on invitro plantlet production and acclimatization of rough lemon (Citrus jambhiri Lush.). M.Sc. Thesis, Punjab Agricultural University, Ludhiana.
Pena, L. and Navarro, L. (1999). Transgenic citrus. In: Y.P.S.(ed.), Biotechnology in Agriculture and Forestry, Vol.44,Transgenic Tress, pp.39-54. Springer-Verlag, Berlin, Germany.
Raman, H., Gosal, S.S. and Brar, D.S. (1992). Plant regen-eration from callus culture of Citrus limon and C jamb-hiri. Crop Improvement.19 (2):100-103.
Rashad, M., Khan, M.M., Ramzan, R., Adnan, S. and Khan F.A. (2005). In vitro regeneration and somatic embryo-genesis in Citrus autantifolia and Citrus sinensis. International Journal of Agriculture and Biology. 7(2): 518-520.
Rattanpal, H.S., Kaur, G. and Gupta, M. (2011). In vitro plant regeneration in rough lemon (Citrus jambhiri Lush) by direct organogenesis. African Journal of Bio-technology, 10(63):13724-13728.
Rezadost, M.H., Sohani, M.M., Hatamzadeh, A. and Reza, M. (2013). In vitro regeneration of sour orange (Citrus aurantium L.) via direct organogenesis. Plant Knowledge Journal. 2(4):150-156 .
Roistacher, C.N., Navarro, L. and Murashige, T. (1976). Recovery of citrus selections free of several viruses, exocortis viroid and spiroplasma citri by shoot-tip graft-ing in vitro. Proceedings 7th Conference International Organization Citrus Virologists, IOCV, Riverside, 186–193.
Saini, H.K., Gill, M.S. and Gill, M.I.S. (2010). Direct shoot organogenesis and plant regeneration in rough lemon (Citrus jambhiri Lush.). Indian Journal of Biotechnol-ogy. 9(4): 419-423.
Silva, R.P., Costa, M.A.P.C., Souza, A.S. and Almeida, W.A.B. (2005). Regeneracao de plantas de laranja-Pera via organogênese invitro. Pesquisa Agropecuaria Bra-sileira. 40(12):1153-1159.
Silva, R.P., Souza, A.J., Mendes, B.M.J. and Mourao Filho, F.A.A. (2010). Sour orange bud regeneration and in vitro plant development related to culture medium com-position and explant type. Revista Brasileira de Fruticultura .32(1):1-8.
Singh, S., Bansal, M.L., Singh, T.P. and Kumar, P. (1998). Statistical Methods for Research Workers. Kalyani Publishers, New Delhi.
Te-Chato, S. and Nudoung, S. (1998). Tissue culture of Citrus reticultata Blanco cv. Shogum and gene transfor-mation by Agrobacteria. Proc. Abstracts IMT-GT UNITE Conf. Songkxhla.
Thorpe, T.D. and Harry, I.S. (1997). Application of tissue culture to horticulture (In: Horticultural biotechnology, edited by A Altaian & M Ziv. Acta Horticulturae. 447:39-49.
Torres, K.C. (1988). Tissue culture technique for horticultural crops (Vam Mostrand Reinhold, New York), 295-298.
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In vitro plant regeneration in rough lemon (Citrus jambhiri Lush.) through epicotyl segments by direct shoot organogenesis. (2016). Journal of Applied and Natural Science, 8(2), 724-729. https://doi.org/10.31018/jans.v8i2.865
