AJPS  Vol.7 No.17 , December 2016
In Vitro Study of Callogenesis and Regeneration Potential of Elite Wheat (Triticum aestivum L.) Cultivars
Abstract: The potential for biotechnological applications in crop improvement programs requires identifying genotypes that allow cell/tissue culture with predictable plant regeneration. In the past, many genotypes of wheat (Triticum aestivum L.) have been examined for potential use in tissue culture studies. The present research work has also been designed to study in vitro callogenesis expression and regeneration potential of wheat cultivars under controlled laboratory conditions. Seeds of four elite commercial high yielding cultivars of wheat namely: NARC-2011, AAS-2011, PAK-2013 and GAL-2013, were collected from the Crop Science Institute National Agricultural Research Center (CSI-NARC) Islamabad, as the source of plant material for in vitro studies. The seeds were surface sterilized in 10% sodium hypochlorite solutions for 10 minutes with continuous shaking under laminar air flow hood. After that seeds were placed on MS (Murashige & Skoog, 1962) based callus induction and regeneration medium with various concentrations of 2, 4-D and BAP in separate test tubes. Maximum callus induction frequency of 90% for Pak-13 and AAS-11, followed by 87% and 83% for Gla-13 and NARC-11, respectively, was recorded at 4 mg/l and 6 mg/l of 2, 4-D. Similarly, maximum regeneration of 90% for AAS-11 and Pak-13, followed by 80% and 87% for NARC-11 and Gla-13 respectively, was recorded on MS basal medium containing 1.5 mg/l of BAP. An increasing trend in regeneration from 0.5 to 1.5 mg/l of BAP was observed but it gradually decreased with increasing concentration of BAP from 1.5 mg/l for all wheat cultivars. The callus formed under light was golden brown, dry nodule and smooth compact and less embryogenic while under dark conditions, it was white to yellowish white, dry nodule and compact and more embryogenic. Best results for callus induction and regeneration were obtained at temperature (24°C ± 1°C) for all wheat cultivars.
Cite this paper: Iqbal, M. , Iqbal Raja, N. , Asif, S. , Ilyas, N. , Hussain, M. , Yasmeen, F. , Ejaz, M. , Asim Sultan, M. , Aslam, S. and Javed, H. (2016) In Vitro Study of Callogenesis and Regeneration Potential of Elite Wheat (Triticum aestivum L.) Cultivars. American Journal of Plant Sciences, 7, 2515-2526. doi: 10.4236/ajps.2016.717219.

[1]   Shewry, P.R. (2009) Wheat. Journal of Experimental Botany, 60, 1537-1553.

[2]   Zale, J.M., Agarwal, S., Loar, S. and Steber, C.M. (2009) Evidence for Stable Transformation of Wheat by Floral Dip in Agrobacterium tumefaciens. Plant Cell Reports, 28, 903-915.

[3]   Alam, M.S., Quayum, M.A. and Islam, M.A. (2011) The Changing Crop Production Practices over the Years: The Mystery of Economics. Bangladesh Journal of Agricultural Research, 36, 657-668.

[4]   Sramkova, Z., Gregova, E. and Sturdik, E. (2009) Chemical Composition and Nutritional Quality of Wheat Grain. Acta Chimica Slovaca, 2, 115-138.

[5]   Bhalla, L.P., Ottonhof, H.H. and Singh, M.B. (2006) Wheat Transformation: An Update of Recent Progress. Euphytica, 149, 353-366.

[6]   Rosegrant, M.W., Sombilla, M.A., Gerpacio, R.V. and Ringler, C. (1997) Global Food Markets and US Exports in 21st Century. The Illinois World Food and Sustainable Agriculture Program Conference, Meeting the Demand for Food in 21st Century: Challenges and Opportunities, 28th May 1997, Urbana.

[7]   GeoHive. (2009) Global Statistics.

[8]   Razzaq, A., Zhiying, M. and Haibo, W. (2004) Genetic Transformation of Wheat (Triticum aestivum L.): A Review. Molecular Plant Breeding, 2, 457-464.

[9]   Hamayun, M., Khan, S.A., Shinwari, Z.K., Khan, A.L., Ahmad, N. and Lee, I.J. (2010) Effect of Polyethyleneglycol Induced Drought Stress on Physio-Hormonal Attributes of Soybean. Pakistan Journal of Botany, 42, 977-986.

[10]   Rashid, H., Khan, M.H., Chaudhry, Z., Bano, A. and Raja, N.I. (2012) An Improved Agrobacterium Mediated Transformation System in Wheat. Pakistan Journal of Botany, 44, 297-300.

[11]   Patnaik, D. and Khurana, P. (2001) Wheat Biotechnology: A Minireview. Electronic Journal of Biotechnology, 4, 94-100.

[12]   Noor, S., Ali, G.M., Rashid, U., Arshad, M., Ali, S. and Zafar, Y. (2009) Optimization of Callus Induction and Regeneration System for Pakistani Wheat Cultivars Kohsar and Khyber-87. African Journal of Biotechnology, 8, 5554-5558.

[13]   Shah, M.M., Khalid, Q., Khan, U.W., Shah, S.A.H., Shah, S.H., Hassan, A. and Pervez, A. (2009) Variation in Genotypic Responses and Biochemical Analysis of Callus Induction in Cultivated Wheat. Genetics and Molecular Research, 8, 783-793.

[14]   Mahmood, I., Razzaq, A. and Khan, Z.U. (2012) Evaluation of Tissue Culture Responses of Promising Wheat (Triticum awstivum L.) Cultivars and Development of Efficient Regeneration System. Pakistan Journal of Botany, 44, 277-284.

[15]   Sarker, R.H. and Biswas, A. (2002) In Vitro Plantlet Regeneration and Agrobacterium-Mediated Genetic Transformation of Wheat (Triticum aestivum L.) Plant Tissue Culture, 12, 155-165.

[16]   Shah, M.I., Jabeen, M. and Ilahi, I. (2003) In Vitro Callus Induction, Its Proliferation and Regeneration in Seed Explants of Wheat (Triticum aestivum L.) var.LU-26S. Pakistan Journal of Botany, 35, 209-217.

[17]   Kabir, A.H., Mahfuz, I., Razvy, M.A., Ahmed, M.B. and Alam, M.F. (2008) Indirect Organogenesis and Somaclonal Variation in Four Rice Cultivars of Bangladesh. Journal of Applied Science Research, 4, 451-458.

[18]   Rahman, M.M., Shamsuddin, A.K.M. and Asad, U. (2008) In Vitro Regeneration from Mature Embryos in Spring Wheat. International Journal of Sustainable Crop Production, 3, 76-80.

[19]   Saad, M., Hamid, R., Yasmin, S. and Minhas, N.M. (2004) Plant Regeneration by Somatic Embryogenesis from Callus of Mature Seed Explants of Bread Wheat (Triticum aestivum L.). Pakistan Journal of Botany, 36, 629-634.

[20]   Benkirane, H., Sabounji, K., Chlyah, A. and Chlyah, H. (2000) Somatic Embryogenesis and Plant Regeneration from Fragments of Immature Inflorescences and Coleoptiles of Durum Wheat. Plant Cell, Tissue & Organ Culture, 61, 107-113.

[21]   Nasircilar, A.G., Turgut, K. and Fiskin, K. (2006) Callus Induction and Plant Regeneration from Mature Embryos of Different Wheat Genotypes. Pakistan Journal of Botany, 38, 637-645.

[22]   Yasmin, S., Khan, I.A., Khatri, A., Seema, N., Nizamani, G.S. and Arain, M.A. (2009) In Vitro Plant Regeneration in Bread Wheat (Triticum aestivum L.). Pakistan Journal of Botany, 41, 2869-2876.

[23]   Jones, H.D. (2005) Wheat Transformation: Current Technology and Applications to Grain Development and Composition. Journal of Cereal Science, 41, 137-147.

[24]   Vendruscolo, E.C.G., Schuster, I., Negra, E.S. and Scapim, C.A. (2008) Callus Induction and Plant Regeneration by Brazilian New Elite Wheat Genotypes. Crop Breeding & Applied Biotechnology, 8, 195-201.

[25]   Iqbal, M., Asif, S., Ilyas, N., Raja N.I., Hussain, M., Shabir, S., Faz, M.N.A. and Rauf, A. (2016) Effect of Plant Derived Smoke on Germination and Post Germination Expression of Wheat (Triticum aestivum L.). American Journal of Plant Sciences, 7, 806-813.

[26]   Murashige, T. and Skoog, F. (1962) A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Plant Physiology, 15, 473-497.

[27]   Satyavathi, V.V., Jauhar, P.P., Elias, E.M. and Rao, B. (2004) Effects of Growth Regulators on in Vitro Plant Regeneration in Durum Wheat. Crop Science, 44, 1839-1846.

[28]   Rashid, U., Ali, S., Ali, G.M., Ayub, N. and Masood, M.S. (2009) Establishment of an Efficient Callus Induction and Plant Regeneration System in Pakistani Wheat (Triticum aestivum) cultivars. Electronic Journal of Biotechnology, 12, 1-12.

[29]   Raja, N.I., Bano, A., Rashid, H., Khan, M.H. and Chaudhry, Z. (2009) Effect of Age of Embryogenic Callus on Plant Regeneration in Local Cultivars of Wheat (Triticum aestivum L.). Pakistan Journal of Botany, 41, 2801-2806.

[30]   Biagioli, C., Rios, R., Basso, A., Franzone, P. and Prina, A. (2006) In Vitro Culture Response of Barley (Hordeum vulgare) Ethylene Synthesis Mutant MC 169. Electronic Journal of Bio-technology, 9, 272-275.

[31]   Ali, A., Muhammd, I., Abdul, M., Naima, H.N., Abdul, R. and Shahid, A. (2013) In Vitro Conservation and Production of Vigorous and Desiccate Tolerant Synthetic Seed Formation in Sugarcane (Saccharum officinarum L.). 47th Annual Conference of Pakistan Society of Sugar Technologists, Rawalpindi Pakistan, 9-10 September 2013.

[32]   Hussain, M., Raja, N.I., Iqbal, M., Iftikhar, A., Sadaf, H.M., Sabir, S., Sultan, M.A. and Faz, M.N.A. (2016) Plantlets Regeneration via Somatic Embryogenesis from the Nucellus Tissues of Kinnow Mandarin (Citrus reticulata L.). American Journal of Plant Sciences, 7, 798-805.

[33]   Hussain, M., Yamin, B., Naveed, I.R., Muhammad, I., Sumaira, A., Nida, T., Muhammad, I. and Anam, I. (2016) A Review of Therapeutic Potential of Ajuga bracteosa: A Critically Endangered Plant from Himalaya. Journal of Coastal Life and Medicine, 4, 918-924.

[34]   Iqbal, M., Aamir, A., Naima, H.N., Umair, A.K., Muhammad, N.A.F., Muhammad, I., Danish, A. and Mubashir, H. (2016) Effect of Explants and Growth Regulators on the Expression of Callogenesis Somatic Embryogenesis and Plantlets Formation in Sugarcane (Saccharum officinarum L.). International Journal of Biosciences, 9, 147-156.