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Development of Doubled Haploid Line Through Anther 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.

Hisham Al Rabbi
Scientific Officer
Biotechnology Division, BRRI Head Office, Gazipur 1701.

Abstract/ Executive Summary:

Anther culture combined with conventional breeding has proved to be an effective tool of regenerating varieties with the desired traits in a shorter period of time. Therefore, anthers of promising F₁s are cultured in the hope of generating potential high yielding early maturity rice lines. Three F₁ population were grown under the optimum fertility and management. Boots with appropriate stage were collected from F₁ plants and pre-incubated at 8ºC for 8-10 days. The panicles were cut into small pieces. They were then surface sterilized and incubated in three different media (KA, KC and KE) and kept in an incubator under dark condition at 25ºC. Calli were then transferred into regeneration medium (MS medium containing 1.0 mg/l NAA+ 1.0 mg/l Kn) and incubated under cool white fluorescent lamps (approximately 1000 lux, at 25ºC) for regeneration. Highest number of callus was obtained from hybrid anthers of BRRI dhan29 X BR1 in KE medium followed by the same hybrid anthers in KA medium. Calli obtained from all the three F₁ populations were incubated in the regeneration medium. The highest numbers of green plants were regenerated from the calli of BRRI dhan29 X BR1 followed by BR6158RWBC2-1-2-1-1-1 X BR28

Keywords: Rice, Double Haploid, Regeneration.

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

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Rice

Objectives:

To develop double haploid line with high yield and short duration.

Methodology:

For this study three F₁ population were grown in the net house under the optimum fertility and management. Boots with appropriate stage (uninucliate/mid uninucleate at reduction division stages) were collected from F₁ plants. The boots were pre-incubated in a refrigerator at 8ºC for 8-10 days. The panicles were opened from the boots and cut into small pieces. They were then surface sterilized by immersing in 70% ethanol for 2-3 minutes and placed onto sterile filter paper in Petri dishes to absorb access alcohol. For callus induction, anthers were incubated in three different media (KA, KC and KE) and kept in an incubator under dark condition at 25ºC until callus initiation and subsequent growth up to appropriate size (2.0mm). Calli were then transferred into regeneration medium (MS medium containing 1.0 mg/l NAA+ 1.0 mg/l Kn) and cultures were incubated in a cultured room under cool white fluorescent lamps (approximately 1000 lux, at 25ºC) for regeneration. Eight weeks after transfer of calli into the regeneration medium, data were taken on number of anther plated, number of callus produced, number of green and albino plant regenerated.

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

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Anthers of three F₁s were plated in three different media (KA, KC and KE). Calli were initiated 2-3 weeks after plating in the callus induction medium. Data were taken for callus induction 8-10 weeks after anthers were plated. Highest number of callus was obtained from hybrid anthers of BRRI dhan29 X BR1 in KE medium followed by the same hybrid anthers in KA medium. Calli obtained from all the three F₁ populations were incubated in the regeneration medium. The highest numbers (27) of green plants were regenerated from the calli of BRRI dhan29 X BR1 followed by BR6158RWBC2-1-2-1-1-1 X BR28.

Knowledge Management: Report/Proceedings