ABB  Vol.4 No.5 , May 2013
Protocol optimization and evaluation of rice varieties response to in vitro regeneration
ABSTRACT

Understanding the control of related molecular mechanisms is impossible without efficient and reproducible regeneration and stable genetic transformation. The key factors for enhancing successful regeneration are genotypes, tissue source of explants, combination and concentration of growth regulators, and culture conditions. In the present study different methods of regeneration were tested in a set of 12 rice accession representing indica, japonica, aromatic and wild groups. The highest frequency of shoot regeneration was achieved using mesocotyls derived from in vitro grown seeds cultured on Murashige and Skoogs, basal medium without growth regulators, when cultured on medium supplemented with Benzyl Adenine (BA) 0.5 mg/l under subdued light at 25°C ± 2°C. Under these conditions mesocotyls (Plurality of meristems) produced 3 to 4 tiller shoots in primary culture. One seed/One single mesocotyls segment produced over 5 to 9 shoots, arising primarily through direct organogenesis after 3 weeks of culture. Through callusing phase different rice cultivars produced different percentage of callusing however Basmati 370 gave high percentage in response to callussing, organogenesis and root formation. In between these two methods, the direct shoot regeneration gave 80% - 100% result besides the wild species in respect with their genotype whereas the indirect organogenesis gave only 10% - 30% of plantlets. So the direct multiplication from mesocotyls is an efficient method for plantlet regeneration of rice cultivars through in vitro culture. Tissue culture derived plants when evaluated with rice markers no variability is found. For regeneration genotypes and its trigger ing factors like medium composition, culture conditions and type of explants are playing a combined role.


Cite this paper
Puhan, P. and Siddiq, E. (2013) Protocol optimization and evaluation of rice varieties response to in vitro regeneration. Advances in Bioscience and Biotechnology, 4, 647-653. doi: 10.4236/abb.2013.45085.
References
[1]   Khush, G.S. and Virk, P.S. (2000) Rice breeding: Achievement and future strategies. Crop Improvement, 27, 115144.

[2]   Hoque, E.H. and Mansfield, J.W. (2004) Effect of genotype and explants age on callus induction and subsequent plant regeneration from root-derived callus of indica rice genotypes. Plant Cell, Tissue and Organ Culture, 78, 217-223. doi:10.1023/B:TICU.0000025640.75168.2d

[3]   Rueb, S., Leneman, M. Schilperoort, R.A. and Hensgens, L.A.M. (1994) Efficient plant regeneration through embryogenesis from callus induced on mature rice embryos (Oryza sativaL.). Plant Cell, Tissue and Organ Culture, 36, 259-264. doi:10.1007/BF00037729

[4]   Ge, X., Chu, Z. Lin Y. and Wang, S. (2006) A tissue culture system for different germplasms of indica rice. Plant Cell Reports, 25, 392-402. doi:10.1007/s00299-005-0100-7

[5]   Khanna, H.K. and Raina, S.K. (1998) Genotype x culture media interaction effects on regeneration response of three indica rice cultivars. Plant Cell, Tissue and Organ Culture, 52, 145-153. doi:10.1023/A:1006032303195

[6]   Al-Forkan, M., Rahim, M.A. Chowdhury, T. Akter, P. and Khaleda, L. (2005) Development of highly in vitro callogenesis and regeneration system for some salt tolerant rice (Oryza sativa L.) cultivars of Bangladesh. Biotechnology, 4, 230-234. doi:10.3923/biotech.2005.230.234

[7]   Kunanuvatchaidach, R., Godwin, I.D. and Adkins, S.W. (1995) High efficiency plant regeneration from callus induced on mature indica rice caryopses. Australian Journal of Botany, 43, 337-348. doi:10.1071/BT9950337

[8]   Sugimoto, K., Miyake, H. and Takiyoka, Y. (2000) Genetic diversity of regeneration ability of Anther culture of rice (Oryza sativa L). Plant Production Science, 3, 387391. doi:10.1626/pps.3.387

[9]   Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiologia Plantarum, 15, 473-497. doi:10.1111/j.1399-3054.1962.tb08052.x

[10]   Chu, C.C. (1978) The N6 medium and its applications to anther culture of cereal crops. In: Proceedings of Symposium on Plant Tissue Culture, 25-30 May 1978, Science Press, Peking, 45-50.

[11]   Duncan, D.B. (1955) Multiple range and multiple F tests. Biometrics, 11, 1-42. doi:10.2307/3001478

[12]   Nishimura, A., Ashikari, M. Lin, S. Takashi, T. Angeles, E.R. Yamamoto, T. and Matsuoka, M. (2005) Isolation of a rice regeneration quantitative trait loci gene and its application to transformation systems. Proceedings of the National Academy of Sciences of USA, 102, 11940-11944. doi:10.1073/pnas.0504220102

[13]   Puhan, P., Vipparla, A. Vemireddy, L.R. Anuradha, G. and Siddiq, E.A. (2012) An efficient and universal Agrobacterium-mediated transformation protocol in rice. Journal of Plant Biochemistry and Biotechnology, 21, 252260.

[14]   Visarada, K.B.R.S. and Sharma, N.P. (2002) Qualitative assessment of tissue culture parameters useful in transformation of indica rice. Current Science, 82, 343-347.

 
 
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