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Development of Drought Tolerance Rice Lines Through in Vitro Screening

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

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

Md Monirul Islam
Senior Scientific Officer
Biotechnology Division, BRRI Head Office, Gazipur 1701.

Abstract/ Executive Summary:

Drought is one of the major environmental stresses during T aman season in Bangladesh affecting the plant growth adversely and reduces the grain yield. About 5.73 million hector of lands of Bangladesh is subjected to moderate to severe drought. Northern part of Bangladesh is the store house of crop production but these area are relatively dry and drought prone. Therefore, attempts have been undertaken for developing suitable drought tolerant variety through in vitro screening with Poly Ethylene Glycol (PEG). Three varieties (BR11, BRRI dhan28 and BRRI dhan29) were used. First mature healthy seeds were dehusked and then surface sterilized with water having 2-3 drops of Tween 20 for 10 minutes. Then this seeds were sterilized with 70% ethanol for 3 minutes followed by 0.1% mercuric chloride for 15 minutes and washed 2-3 times with sterile distilled water. About 100-120 sterilized seeds of each genotype for each treatment were placed into callus induction medium (CIM) containing MS basal salts, 2.0 mg/L, 2,4-dichlorophenoxyacetic acid (2,4-D) and 30 g/L sucrose. Three levels (5.0, 10.0 and 15.0g/L) of polyethylene glycol (PEG) were used as stressing agent in CIM media. Callus developed at the swollen junction of radicle and mesocotyl were excised on the 10^th day and cultured on the same medium. Then this calli were incubated for another 20 days in the dark. After a month, induced calli were transferred to callus multiplication medium (CMM) consisting MS basal salts, 1 mg l^-1 2,4-D 2,4-D and 3 different concentration of PEG (5.0, 10.0 and 15.0g/L). These calli were kept in the dark for 21 days for proliferation. Then calli from individual seeds were sub cultured on freshly prepared MS with 1mg/L 2,4-D without any PEG for 21 days in the dark for proliferation of putative drought-tolerant cells. Finally, calli were transferred to regeneration medium (MS with 2 mg l^-1 Kinetin , 0.5 mgl^-1 BA, and 0.5 mgl^-1 NAA). After 3 weeks green spots were appeared on the callus surface and within 1 month plantlets were regenerated. Callus production was varied in all the genotypes tested under different concentration of PEG. About 129 putative stress tolerant calli were obtained from using PEG 6000 @ 15g/L.

Keywords: Rice, Drought tolerence, PEG

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

Start Date: 01-07-2011

End Date:

Program Area: Variety and Species

Commodity: Drought

Objectives:

To develop drought tolerant rice variety

Methodology:

Three varieties (BR11, BRRI dhan28 and BRRI dhan29). First mature healthy seeds were manually cleaned and dehusked. Dehusked seeds were then surface sterilized with water having 2-3 drops of Tween 20 for 10 minutes. Then this seeds were sterilized with 70% ethanol for 3 minutes followed by 0.1% mercuric chloride for 15 minutes. Finally sterilized seeds were washed 2-3 times with sterile distilled water. About 100-120 sterilized seeds of each genotype for each treatment were placed into callus induction medium (CIM) containing MS basal salts, 2.0 mg/L, 2,4-dichlorophenoxyacetic acid (2,4-D) and 30 g/L sucrose. Three levels (5.0, 10.0 and 15.0g/L) of polyethylene glycol (PEG) were used as stressing agent in CIM media. Callus developed at the swollen junction of radicle and mesocotyl were excised on the 10^th day and cultured on the same medium. Then this calli were incubated for another 20 days in the dark. After a month, induced calli were transferred to callus multiplication medium (CMM) consisting MS basal salts, 1 mg^-1 2,4-D 2,4-D and 3 different concentration of PEG (5.0, 10.0 and 15.0g/L). These calli were kept in the dark for 21 days for proliferation. Then calli from individual seeds were sub cultured on freshly prepared MS with 1mg/L 2,4-D without any PEG for 21 days in the dark for proliferation of putative drought-tolerant cells. Finally, calli were transferred to regeneration medium (MS with 2 mg l^-1 Kinetin , 0.5 mg l^-1 BA, and 0.5 mgl^-1 NAA). After 3 weeks green spots were appeared on the callus surface and within 1 month plantlets were regenerated.

Source of Information: Proceedings of the BRRI Annual Internal Review for 2011-2012

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Callus production was varied in all the genotypes tested under different concentration of PEG. About 129 putative stress tolerant calli were obtained from using PEG 6000 @ 15g/L. After plant regeneration materials will be screened for drought tolerance and used as breeding materials for varietal development.

Knowledge Management: Report/Proceedings