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Establishment of Regeneration Protocol for BRRI Rice Varieties Through Seed Culture

Enamul Hoque
Principal Scientific Officer
Biotechnology Division, BRRI Head Office, Gazipur 1701.

Md Shamsher Ali
Chief Scientific Officer
Biotechnology Division, BRRI Head Office, Gazipur 1701.

Nasrin Sultana
Scientific Officer
Biotechnology Division, BRRI Head Office, Gazipur 1701.

Abstract/ Executive Summary:

The major constraints for in vitro culture of Indica rice are the low rates of callus production and subsequent plant regeneration. The effect of different salts at different concentration in the induction media to enhance callus induction and subsequently plant regeneration frequency from mature seed was tried. Fourteen BRRI released rice varieties were used. Mature dehusked seeds were sterilized and cultured in callus induction medium (MS containing 2.0mg/L 2, 4-D) and with 6 different concentrations (0.0, 1.0, 2.0, 3.0, 4.0 and 5.0 g/l) of two different salts (NaCl, and Na₂SO₄) . Embryogenic calli were placed into regeneration medium (MS medium with 1.0 mg/l NAA+ 1.0 mg/l Kn). On an average callus induction was higher in the medium with supplemental NaCl and lowest in the medium containing Na₂SO₄. Generally callus induction frequency reduced as salt loading increased. Higher concentration of salts (5.0 g/L) reduced the callus induction to about one third or less indicating the deleterious effect of salts on callusing. Among the rice variety tested, BRRI dhan41 was recognized as the highest callus producing variety and BRRI dhan36 was identified as the lowest. Calli of BRRI dhan38 obtained from callus induction medium containing NaCl showed higher regeneration response.

Keywords: Rice, Regeneration, Seed culture.

Location: Biotechnology Division, BRRI Head Office, Gazipur 1701.

Start Date: 01-07-2007

End Date:

Program Area: Variety and Species

Commodity: Rice

Objectives:

To identify the regeneration ability of BRRI released rice varieties using different salts.

Methodology:

Fourteen BRRI released rice varieties (BRRI dhan32 to BRRI dhan43 and BRRI dhan45 to BRRI dhan46) were used for this study. Mature healthy dehusked seeds were sterilized first by 2-3 drops of Tween 20 for 10 minutes. Then the seeds were sterilized with 70% ethanol for 3 minutes and then with 0.1% mercuric chloride for 20 minutes, followed by 5-6 washing with sterile distilled water. Sterile seeds were then cultured in Petri dish containing callus induction medium (MS containing 2.0mg/L 2, 4-D) and with 6 different concentrations (0.0, 1.0, 2.0, 3.0, 4.0 and 5.0 g/l) of two different salts (NaCl, and Na₂SO₄). The Petri dishes were sealed with Para film and incubated at 25±1°C for four weeks before callus induction responses were assessed. After four weeks of incubation, data were taken on callus induction frequency. For regeneration, eight-week-old embryogenic calli were placed into Petri dishes containing regeneration medium (MS medium supplemented with 1.0 mg/l NAA+ 1.0 mg/l Kn). All such calli were kept at 25±1°C under continuous light from fluorescent lamps. Data on shoot regeneration were collected eight weeks after transfer of calli.

Source of Information: Proceedings of the BRRI Annual Internal Review for 2007-2008

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The study revealed that both salts and concentration of the salts had great impact on callus induction and plant regeneration. On an average callus induction was higher in the medium with supplemental NaCl and lowest in the medium containing Na₂SO₄. Higher concentration of salts (5.0 g/L) reduced the callus induction to about one third indicating the deleterious effect of salts on callusing. BRRI dhan41 was recognized as the highest callus producing variety and BRRI dhan36 was identified as the lowest. In this study, plant regeneration was found to be very low, even some of the variety did not produce any plantlets. Low in vitro plant regeneration is very common in Indica rice. Our results also revealed that salts had a deleterious effect on plant regeneration. Calli of BRRI dhan38 obtained from callus induction medium containing NaCl showed higher regeneration response. It is followed by BRRI dhan37 calli which were induced from both NaCl and Na₂SO₄containing medium. Regeneration was reduced as salt concentration is increased compared to control, although 1 gm/l salt treatment showed superiority over other treatments. More studies are needed to find out a suitable media composition containing salt for elevated in vitro response.

Knowledge Management: Report/Proceedings