European Journal of Biotechnology and Bioscience

Authors
Subhabrata Paul, Biswajit Ghosh

Abstract
An efficient propagation procedure for seven wild aromatic rice varieties in West Bengal was developed by using seed germination and the genetic stability of the micropropagated plants was verified by randomly amplified polymorphic DNA (RAPD) analysis. A DNA bank of eighteen aromatic rice varieties was done and stored at -80 ºC. Seedlings were obtained on Murashige and Skoog (MS) medium with 0.70% agar after one week of culture and used as a explants. Different concentrations of cytokines i.e. 6-benzylaminopurine (BAP), 6-γ-γ-[Dimethylallylamino]-purine (2iP) and kinetin (Kn) were used for the next subculture. After 40 d of culture, multiple shoots were obtained. The highest number of shoot, 6.45 ± 0.28 in Kataribhog and 6.50 ± 0.90 in Valuduboraj was observed on MS media supplemented with 3 mg/L BAP and 5 mg/L 2iP respectably. For in vitro conservation purpose, Kataribhog was conserved for maximum time at about 157 d on 5 mg/L 2iPwhich was an impressive results over the all tested media. After profuse rooting was obtained complete plants were successfully transferred to soil and grown up to maturity. Further, colonel fidelity assessment of the micropropagated plants was carried out using RAPD markers analysis. 19 out of 25 arbitrary decamer primers amplified a total of 79reproducibledistinct DNA fragments ranging from 350 to 1,300 bp. Comparison of the bands with the mother plant revealed the monomorphic nature. A result of RAPD markers analysis indicates all the regenerates are true-to-type and there were no somaclonal variations among these regenerates. The above Micropropagation protocol will be useful for long term in vitro conservation and maintenance of genetically uniform aromatic rice plants micropropagated in the lab. This also involves a range of projects which have utilized the DNA bank for further studies and can provide a key resource to exploit the capabilities in the field of biotechnology.