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Improvement of in Vitro Plant Regeneration System of Aromatic and Fine Grain Rice Varieties

A K M Ruhul Baksha
Senior Scientific Officer
Biotechnology Division, Bangladesh Rice Research Institute, Gazipur.

Md. Enamul Hoque
Senior Scientific Officer
Biotechnology Division, Bangladesh Rice Research Institute, Gazipur.

Jannatul Ferdous
Scientific Officer
Biotechnology Division, Bangladesh Rice Research Institute, Gazipur.

Md. Shamsher Ali
Principal Scientific Officer
Biotechnology Division, Bangladesh Rice Research Institute, Gazipur.

Abstract/ Executive Summary:

The study was carried out to evaluate the effect of salts in callus induction and subsequent plant regeneration of five aromatic and fine grain rice varieties. Calli were initiated on MS medium supplement with 2 mg/l 2, 4-D, 20 g/l with three different salts (NaCl, NaHCO₃ and NaSO₄) at four levels of concentrations (2.5, 5.0, 7.5 and 10.0 g/l). Addition of 2.5 g/l NaCl to callus induction medium enhanced callus formation frequency compare to those of NaHCO₃ and NaSO₄. BR5 had the highest frequency of callus production ability (100%) followed by Hassan Saraie (85%), Kararibhog (83%), Kalijira (80%) and BRRI dhan28 (75%). Medium containing high salt concentration (7.5-10.0 g/l) was found to be detrimental for callus induction as well as green plant regeneration response. Average plant regeneration response was found to be better from the medium containing 2.5 g/l NaCl followed by 2.5 g/l of each NaHCO₃ and NaSO₄. BR5 had the highest frequency of plant regeneration ability (90%) followed by Kalijira (85%), Kararibhog (75%) , BRRI dhan28 (75%) and Hassan Saraie (73%) at 2.5 g/l NaCl.

Keywords: Rice, Salt stress, Callus induction, In vitro plant regeneration

Location: Biotechnology Lab, BRRI, Gazipur

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Rice

Objectives:

Find out the effect of different salts concentration on callus induction and subsequent plant regeneration of aromatic and fine grain rice varieties

Methodology:

Five aromatic and fine rice varieties viz Hassan Saraie, Kalijira, Kataribhog, BR5 and BRRI dhan28 were used in the study. Among them BR5 and BRRI dhan28 were released as a modern varieties from Bangladesh Rice Research Institute (BRRI), Bangladesh. Hassan Saraie was collected from Iran and Kalijira and Kataribhog are locally cultivated aromatic rice variety. Mature healthy seeds were first selected and manually dehusked. Seeds were then sterilized by 70% ethanol for 3 munites followed by 0.1% mercuric chloride for 20 minutes with gentle shaking. Sterilized seed s were washed for 5-6 times with sterile distilled water and seeds were then dried with sterilized filterb paper. Media for callus induction based on the formulation of MS basal salts supplemented with 2 mg/l 2, 4-D, 20 g/l sucrose and 8 g/l agar. Three different salts (NaCl, NaHCO₃ and NaSO₄) at four different levels of concentrations were used to observe the effect on callus induction rates. Compositions of callus induction media were as follows: Control (0 salt)= T₀, NaCl supplemented media (T₁= 2.5 g/l, T₂= 5.0 g/l, T₃= 7.0 g/l and T₄= 10.0 g/l of NaCl, respectively), NaHCO₃ supplemented media (T₁= 2.5 g/l, T₂= 5.0 g/l, T₃= 7.0 g/l and T₄= 10.0 g/l of NaHCO₃, respectively), and NaSO₄ supplemented media (T₁= 2.5 g/l, T₂= 5.0 g/l, T₃= 7.0 g/l and T₄= 10.0 g/l of NaSO₄, respectively). The pH of the medium was adjusted to 5.8 before autoclaving. The media was autoclaved for 20 minutes under 121º C and 22-25 ml autoclaved media was poured into 6 cm diameter Petri dishes. Sterilized seeds were plated in the callus induction media and incubated for four weeks at 25 º C under dark condition. Six seeds were plated in each Patri dish and 240 seeds were used for each variety . The frequency of callus induction was calculated by dividing the total number of seeds produced callus by the total number of seeds cultured and multiplied by 100. For plant regeneration, MS media supplemented with 40 g/l sucrose, 1 mg/l NAA, 1 mg/l kinetin and 8 g/l agar were used. Same salts and concentrations were used to evaluate its effect on regeneration response. Four weeks old calli were transferred on to regeneration medium containing different salts. Six to eight weeks old plants were then transferred to small pot for hardening. After three weeks, plants were further transferred to big earthen pots and let them grown until maturity. A complete Randomize Design (CRD) with six replications per treatment for each variety were used analyzed statistically.

Source of Information: Molecular Biology and Biotechnology Journal. 4 (1&2):45-48. 2006

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

2.5 g/l NaCl salt concentration in the both callus induction and regeneration medium have beneficial effect on callus induction and plant regeneration.

Knowledge Management: Journal