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Establishment of the in Vitro Regeneration Protocol for BRRI Rice Varieties Using Different Salts

Md. Shamsher Ali
Chief Scientific Officer and Head
Biotechnology Divition Bangladesh Rice Research Institute Gazipur-1701

Md. Enamul Hoque
Senior Scientific Officer
Biotechnology Divition Bangladesh Rice Research Institute Gazipur-1701

AKM Ruhul Baksha
Senior Scientific Officer
Biotechnology Divition Bangladesh Rice Research Institute Gazipur-1701

Jannatul Ferdous
Scientific Officer
Biotechnology Divition Bangladesh Rice Research Institute Gazipur-1701

Abstract/ Executive Summary:

A study was carried out to find out the effect of different salts in callus induction and subsequent plant regeneration from twenty BRRI released varieties. The study revealed that callus induction and regeneration were badly affected by KCL and least affected by Na₂SO_4. Different concentrations of salts have also influenced callus induction and regeneration ability. Higher concentration of salt (7.5g/l) had deleterious effect on callus induction and regeneration. Among the rice variety tested, BR4 was the highest callus producing variety followed by BR10, respectively. Average highest plant regeneration was observed in BR8 (14.54%) followed by BR10 (13.28%) and BR6 (10.9%) respectively. In some cases, regeneration response was not observed at all. Besides the genotypic effects, the concentration of different salts in callus induction media had also influenced the regeneration frequency. Lower concentrations (2.5- 5.0 g/l) of Na₂SO₄ salts in callus induction medium seemed non-deleterious for callus formation and subsequent plant regeneration. Thus, a suitable salt and its level of concentration in commonly used media need to be identified for good callus induction and subsequently plant regeneration.

Keywords: Seed culture, Salt concentration, Plant regeneration frequency, Seed embryos, BRRI released varieties

Location: Biotechnology Division, BRRI, Gazipur-1701

Start Date: 01-07-2005

End Date: 30-06-2006

Program Area: Variety and Species

Commodity: Rice

Objectives:

To enhance callus induction and subsequently plant regeneration frequency from mature seed embryos of BRRI released varieties.

Methodology:

Dehusked seeds from the rice varieties were used for callus induction. MS medium was selected as basal medium for callus induction and plant regeneration. Rice seeds were manually dehusked and washed with 70% ethanol for 30 seconds. After that the seeds were rinsed in 0.2% HgCl₂ solution for 10 minutes. The materials then washed 3 times with autoclaved distilled water. Finally the materials were kept in tissue paper to soak the residual water. Surface sterilized seeds were cultured with the help of sterilized in forceps into the conical flask containing 25 ml of callus inducing medium supplemented with the 1 mg/L 2,4 D. Calli were initiated on MS medium supplemented with 2.0 mg/l 2.4-D with three different salts (NaCL, Na₂SO₄and KCL) at four different levels of concentrations (0, 2.5, 5.0, 7.5 g/l). In each flask, 10 seeds were inoculated and for each treatment, 30 seeds were cultured. The cultures were kept in the dark for one month at 25 + 1°C, and then they were transferred into the plant regeneration medium fortified with 1 mg/L NAA and 1 mg/L Kinetin. and kept under 16 hour’s photoperiods at 25 + 1°C. Within one month greenish plantlets were obtained from the plated calli.

Source of Information: Proceedings of the BRRI Annual Internal Review for 2005-2006

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Lower concentrations (2.5- 5.0 g/l) of Na₂SO₄ salts in callus induction medium seemed non-deleterious for callus formation and subsequent plant regeneration.

Knowledge Management: Report/Proceedings