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Effect of Plant Growth Regulators on Callus Induction and Plant Regeneration in Anther Culture of Rice

Md. Monirul Islam
Agrotechnology Discipline, Khulna University, Khulna-9208, Bangladesh.

Sanjay Kumar Adhikary
Agrotechnology Discipline, Khulna University, Khulna-9208, Bangladesh.

Purnendu Gain
Agrotechnology Discipline, Khulna University, Khulna-9208, Bangladesh.

Md. Mizanur Rahman
Agrotechnology Discipline, Khulna University, Khulna-9208, Bangladesh.

Noor-e-Alam Siddique
Agrotechnology Discipline, Khulna University, Khulna-9208, Bangladesh.

Abstract/ Executive Summary:

An experiment was conducted to find out the effects of different concentrations and combinations of growth regulators viz. 2, 4 D, IAA, ?-NAA and Kinetin on callus induction from the anthers of a commercial hybrid rice line IR -69690, developed by Bangladesh Rice Research Institute and subsequent plant regeneration. N₆ medium was used as basal medium. Callus induction frequencies in different media combinations ranged from 1.2 to 35.5%. The medium supplemented with 2,4 D 1 mg L^-1, ?-NAA 2 mg L^-1 and Kinetin 1 mg L^-1 was found most effective for callus induction (35.5%). Regeneration of plants from the callus on agarified MS medium supplemented with ?-NAA 0.5 mg L^-1 and Kinetin 3 mg L^-1 was also variable and ranged from 16.7 to 69.3%. Calli derived from the media supplemented with 2,4 D 1 mg L^-1, ?-NAA 2 mg L^-1 and Kinetin 1 mg L^-1 also showed better performance for plant regeneration (69.3%) and among these plants 56.14% were green. However the callus induction medium containing ?-NAA 1 mg L^-1 and Kinetin 1 mgL^-1directly produced green regenerated plants in higher frequency (70%), without transferring the calli on to the regeneration medium; but rate of callus induction in this medium was very low (6%).

Keywords: Hybrid rice, Anther, Callus, Plant regeneration, Haploid

Location: Laboratory of Genetics and Plant Breeding of Agro-technology Discipline, Khulna University

Start Date: 00-00-2001

End Date: 00-00-2001

Program Area: Crop-Soil-Water Management

Commodity: Rice

Objectives:

To study the ability of a commercial hybrid rice line under study in callus induction from anthers and subsequent plant regeneration.
To determine the most suitable combination and concentration of growth regulators that enhances callus induction and plant regeneration.

Methodology:

The experiment was conducted in 2001 in the laboratory of Genetics and Plant Breeding of Agro-technology Discipline, Khulna University, A commercial hybrid rice variety (IR-69690) developed by BRRI was used as source plant for anthers. The panicles were collected from the demonstration plot located at Dumuria Upazilla, Khulna. At the late uninucleate stage (when the distance between the base of the flag leaf and the auricle of the last leaf was 3-6 cm.), the panicles were collected between 8 and 10am and treated with clays at 4-8°C in sealed polythene bags in order to enhance callus production. Cold treated panicles were thoroughly washed in tap water and were placed in the laminar flow cabinet. The panicles were then surface sterilized by immersing them in 70% ethanol for 20 seconds followed by immersing in 0.2% HgCl, solution for 10 min. The treated materials washed 3 times with sterile distilled water. Only those panicles were selected in which maximum anthers had reached 50% length of the spike lets and were green in colour. Fifty to sixty spike lets were taken at a time on sterile petridish and when surface was dried, individual spike lets were cut at the base to free the anthers from the filaments. After that with the help of sterile forceps and needle the anthers were plated to conical flask containing appropriate nutrient medium. The cultures were incubated in the dark at 26±1°C for one month and then under 16 hours photo periods at about 3000 Lux.The induced calli were transplanted into test tubes containing MS medium 'P' supplemented with cx-NAA 05 mg L^-1, Kinetin 3 mg L -1, 50 g L^-1 sucrose, 8 g L^-1 agar. The plated calli were incubated in a growth chamber at 26±1 °C with 16 h of light, at a light intensity of about 2000 Lux. pH of the media both for callus induction and plant regeneration was adjusted to 5.6. In the 7th week after inoculation of anthers, callus induction frequency was calculated on the basis of the number of anthers producing callus. Regenerated plants were counted on the basis of the number of callus producing plant lets. The frequency of callus induction and plant regeneration were calculated as below:

Callus induction frequency (%) = (No. of anthers producing callus/No. of anthers plated) x 100

Plant regeneration frequency (%)= (No. of calli regenerated plantlets /No. of calli plated for regeneration) x 100

Analysis of Variance (ANOVA) was done to determine the effect of different combinations and concentrations of the selected growth regulators on callus induction.

Source of Information: Pakistan Journal of Biological Sciences 7 (3): 331-334, 2004 (ISSN 1028-8880)

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The commercial hybrid rice line (IR-69690) was found to be competent for anther culture study and by the addition of 2, 4 D 1 mg L^-1, with ?-NAA 2 mg L^-1 and Kinetin 1 mg L^-1 to callus induction medium, a higher callusing and plant regeneration response could be obtained.

Knowledge Management: Journal