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Mature Embryo-based in Vitro Regeneration of Indica Rice Cultivars for High Frequency Plantlets Production

H N Barman
Plant Physiology Division, Bangladesh Rice Research Institute (BRRI), Gazipur 1701, Bangladesh

M E Hoque
Biotechnology Division, Bangladesh Rice Research Institute (BRRI), Gazipur 1701, Bangladesh

R K Roy
Biotechnology Division, Bangladesh Rice Research Institute (BRRI), Gazipur 1701, Bangladesh

P L Biswas
Plant Breeding Division, Bangladesh Rice Research Institute (BRRI), Gazipur 1701, Bangladesh

M A I Khan
Plant Pathology Division, Bangladesh Rice Research Institute (BRRI), Gazipur 1701, Bangladesh

M O Islam
Department of Crop Botany

Abstract/ Executive Summary:

The study was conducted at Biotechnology Division of Bangladesh Rice Research Institute (BRRI) to investigate the effects of plant growing medium and plant growth regulator (PGR) for the callus induction and high frequency plantlets regeneration of indica rice. Ten indica rice varieties viz. BR5, BR11, BRRI dhan28, BRRI dhan29, BRRI dhan33, BRRI dhan41, BRRI dhan47, BRRI dhan48, BRRI dhan49 and BRRI dhan50 were cultured on MS, N6 and LS media. The MS medium was found better for callus induction as compared to N6 and LS media. Among the tested varieties BRRI dhan48 induced the highest percent and best quality callus. Interaction effects of BRRI dhan48 to MS medium yielded 71.55% callus induction. The regeneration efficiency of BRRI dhan48 was tested on MS medium supplemented with different combination of NAA plus BAP and NAA plus kinetin. MS medium supplemented with2.0 mg L^-1 NAA and 2.0 mg L^-1 Kn was found the best in respect of percent regenerated (76.67%) plantlet as well as for the growth of plantlets in vitro.

Keywords: Indica rice (Oryza sativa L.), Growing media, Growth regulators, Callus induction, Regeneration

Location: Biotechnology Division of BRRI, Gazipur, Bangladesh.

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Rice

Objectives:

To determine the Mature Embryo-Based in vitro Regeneration of Indica Rice Cultivars for High Frequency Plantlets Production

Methodology:

The experiment was conducted at Biotechnology Division of BRRI, Gazipur, Bangladesh. Ten BRRI released indica rice varieties viz BR5, BR11, BRRI dhan28, BRRI dhan29, BRRI dhan33, BRRI dhan41, BRRI dhan47, BRRI dhan48, BRRI dhan49 and BRRI dhan50 were used. Three culture media viz MS (Murashige and Skoog, 1962), N6 (Chu et al., 1975) and LS (Linsmaier and Skoog, 1965) were evaluated to select the best medium for callus induction and regeneration. The experiment was conducted in 18 mm × 1.2 mm test tubes and laid out following a completely randomized design (CRD) with five replications. To induce callus, dehusked mature embryo were cultured on MS, N6 and LS media and incubated in growth room at 25±2°C under 12 hr. photoperiod of 50 µmol m-2s-1 provided by florescent tubes. The basal media were supplemented with 30.0 g L^-1 sucrose, 0.3 g L^-1 casamino acid and 2.0 mg L-1 2, 4-Dichlorophenoxyacetic acid (2, 4-D) and solidified with 7.0 g L^-1 agar. The pH of the media was adjusted to 5.8 prior to autoclave at 121°C with 1.16 kg cm^-2 pressure for 20 minutes. The data on callus induction and plantlet regeneration were collected on 21 and 50 days after culture. Callus induction, regeneration, number of plantlets callus^-1, average plantlet height, number of root plantlet^-1, average root length etc were also collected. Data were analyzed to compare mean values to express treatments effects. The means were calculated and analysis of variance of all the characters was performed by F-test. Significance of the difference between the pair of means wasevaluated by Duncan’s Multiple Range Test (Gomez and Gomez, 1984) at the 5% level of significance using MSTAT-C computer programmed (Russel, 1986).

Source of Information: Bangladesh Rice J. 20 (2) : 81-87, 2016

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In present study MS medium gave better callus induction as compared to LS and N6 media respectively. Percent regenerated plantlet was found highest in BRRI dhan48 when cultured in MS medium supplemented with NAA and Kn compared to MS medium supplemented with NAA and BAP. Thus this finding revealed that Kn was more effective than BAP for regeneration and organogenesis. The present result might be helpful in future research on genetic transformation as well as in selection of stress tolerant cultivar development programme. From the result it might be concluded that MS medium for callus induction and MS medium supplemented with 2.0 mg L^-1 NAA and 2.0 mg L^-1 Kn for plantlet regeneration may be recommended for future research work.

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