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Regeneration Efficiency and Genotypic Effect of 15 Indica Type Bangladeshi Rice (oryza Sativa L.) Landraces

Mohammad Khalequzzaman
Genetic Resources and Seed Division, Bangladesh Rice Research Institute, Gazipur-1701.

Nazmul Haq
Department of Civil and Environmental Engineering, University of Southampton, Southampton, SO17 1BJ, UK

Md. Enamul Hoque
Department of Civil and Environmental Engineering, University of Southampton, Southampton, SO17 1BJ, UK

Tamal Lata Aditya
Department of Civil and Environmental Engineering, University of Southampton, Southampton, SO17 1BJ, UK

Abstract/ Executive Summary:

Genotypic effect was observed for embryogenic callus induction and subsequent plant regeneration in 15 indica type Bangladeshi rice (Oryza sativa L.) landraces. Different growth regulators were found to influence callus induction ability and green plant regeneration. The highest frequency (55.6%) of shoot regeneration was observed in Hashikalmi with the supplementary hormone concentrations and combination of 2 mg/l BA, 1 mg/l NAA and 1.5 mg/l Kn in MS. The embryogenic calli derived from mature seeds successfully produced shoots, and rooted well on MS basal salt with IAA. Five landraces which could be utilized in further manipulation, such as transformation and in vitro hybridization, were found to have higher regeneration ability (± 50%). In vitro regenerated plants were successfully established in soil and produced fertile seeds.

Keywords: Mature embryos, Genotypic variability, Embryogenic callus, Plant regeneration, Indica rice

Location: Bangladesh Rice Research Institute (BRRI), Gazipur-1701, Bangladesh

Start Date: 00-00-2000

End Date: 00-00-2001

Program Area: Variety and Species

Commodity: Rice

Objectives:

To establish any efficient in vitro plant regeneration system for future use in genetic transformation and in vitro fusion studies.

Methodology:

Experiments were conducted at the Environmental Science Laboratory of the University of Southampton, UK during 2000 and 2001. Fifteen rice landraces received from Bangladesh Rice Research Institute (BRRI), Gazipur-1701, Bangladesh were included in this study. The landraces were collected from different ecotypes and rainfall distribution pattern of Bangladesh. Mature and healthy seeds were selected and dehusked. They were first surface sterilized in 70% (v/v) ethanol for three minutes with gentle shaking. Thereafter, they were washed with sterilized distilled water for three times. Then the seeds were again surface sterilized in 50% (v/v) commercial bleach (approx. 5% NaOC1) for 30 minutes with gentle agitation and subsequently rinsed with sterilized distilled water for five times. MS basal salt supplemented with 3% sucrose (w/v) (Sigma); 1.5 - 2.5 mg/l, 2,4-D, 50 mg/l nicotinic acid and solidified with 0.5% agarose (w/v) (Sigma) was used to find out the callus induction ability of the genotypes. Three different concentrations viz. 1.5, 2 and 2.5 mg/l of 2,4-D were used in the media to evaluate their effect on callus induction ability. The percentage of plant regeneration response for each genotype was calculated dividing total number of calli produced plantlets by the total number of calli cultured and multiplied by 100. A completely randomized design with three replications per treatment for each genotype was followed in this study. ANOVA was integrated for assessment of the variation due to genotypes and media. LSD test was used to show the variation between mean values of different parameters. Standard error of means was also calculated.

Source of Information: Plant Tissue Cult. 15(1): 33-42, 2005 (June)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The present screening test results showed that out of 15 landraces, two failed to produce green plantlets from callus and the rest produced plantlets with different rate of success, which clearly indicated the genotypic effect of plant regeneration. The identification of genotype(s) with superior culture ability is a prerequisite for further experiments, such as transformation, in vitro hybridization and somatic hybridization. It appears from the present study that at least five indica rice landraces were found to be best for embryogenic callus induction and green plant regeneration. Such type of finding will prove useful for their further manipulation. The embryogenic calli derived from mature seeds successfully produced shoots, and rooted well on MS basal salt with IAA. Five landraces which could be utilized in further manipulation, such as transformation and in vitro hybridization, were found to have higher regeneration ability (± 50%). In vitro regenerated plants were successfully established in soil and produced fertile seeds.

Knowledge Management: Journal