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Callus Induction and Plant Regeneration from Mesocotyl Explant of Rice (oryza Sativa L.)

M Asaduzzaman
Biotechnology Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh

M Rahman
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh

M A Bari
Biotechnology Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

Somatic embryos were obtained from mesocotyl derived callus of Oryza sativa L. High frequency of embryogenic cultures, growth regulator supplement and incubation conditions during the callusing phase were found critical. For induction of callus from mesocotyl N₆ medium was supplemented with different concentrations of 2, 4 - D. The best callusing was obtained from explants cultured on Ns medium containing 2.5 mg/ l 2, 4 - D ranging from 80% in BR - 31, 75% in BR - 30 and 50% in BR - 28. Shoot regeneration was achieved after transferring the callus to MS medium supplemented with BAP and NAA. Mesocotyl derived calli from BR - 30 produced the highest number (4.57) of shoots and percentage of shoot formation was 67%. The media without any growth regulators failed to produce any shoots from the callus.

Keywords: Rice, Callus induction, Regeneration, Mesocotyl

Location: Regional Rice Research Institute, Rajshahi

Start Date: 01-01-1994

End Date: 30-06-1995

Program Area: Variety and Species

Commodity: Rice

Objectives:

To study the three rice varieties and their response to callus induction and regeneration.

Methodology:

Seeds of three varieties of rice, BR - 28, BR - 30 and BR - 31 were obtained from Regional Rice Research Institute of Rajshahi, Bangladesh. Seeds of rice varieties were carefully dehusked and they were surface sterilized with 70% ethyl alcohol for five minutes. Sterilized seeds were washed several times with autoclaved distilled water. Washed seeds were again sterilized with sodium hypochloride (NaOCI) for five minutes and washed several times with autoclaved distilled water. The sterilized seeds of different varieties were sown on conical flax containing MS medium and kept at 25 ± 1°C in dark for 7 days. Relatively hard and swollen base just above the seed its scutellum and leaf base is designated as mesocotyl and used as the tissue culture explants namely scutellum, leaf base, mesocotyl and root were also considered as suitable explants in rice tissue culture studies. Mesocotyls were carefully excised from the seedling and cultured on three callus inducing media viz. Ns, LS and MS containing different concentrations of 2, 4 - D. The cultured mesocotyls in petridish were incubated in dark condition at 25 ± 1 °C for about three to four weeks in order to induce callus formation. Calli were formed within three to four weeks and they were regularly sub cultured at 15 days interval. Six week old mesocotyl derived calli were transferred to three regeneration media (MS + BAP 0.5mg/ 1 + NAA 0.05 mg/ I, N₆ + BAP 0.5 mg/ 1 + NAA 0.05 mg/ 1 and 2 mg/ 1 BAP + 0.5 mg/ 1 NAA) and kept at 25 ± 1°C in the light. Mesocotyl derived calli cultured on regeneration media were kept for three to four weeks. After two to three weeks the calli began to initiate shoot as well as root in the same media. The plantlets regenerated from calli cultured in conical flax would become complete plants having good number of green leaves and roots. Plantlets were successfully transferred to the plastic vessel containing sterilized soil.

Source of Information: JOURNAL OF BIO-SCIENCE, THE INSTITUTE OF BIOLOGICAL SCIENCES RAJSHAHI UNIVERSITY, Vol. 13, 2005, pp. 23-26, ISSN 1023–8654.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present investigation it was observed that different concentrations of 2, 4 - D alone produced callus in high frequency from mesocotyl explants of rice. It is also noted that, in the present study, there were no albino plants obtained from mesocotyl derived callus. Albino plants are unwanted and their production greatly limits the objects of tissue culture technique. Among the three rice varieties studied in this investigation BR - 28 and BR - 31 failed to perform a complete process of artificial regeneration under mesocotyl culture. However BR - 30 experienced an efficient performance under both the purviews of callus induction and regeneration in mesocotyl culture. The present study revealed that the variety BR - 30 proved to be a suitable rice variety holding potentiality of genetic manipulation to be applied in mesocotyl explant culture.

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