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Identification of Callus Induction and Plant Regeneration Responsiveness in Presence of Nacl in In Vitro Culture of Some Deepwater Rice (oryza Sativa L.) Cultivars

L. Khaleda
Department of Genetic Engineering, University of Chittagong, Chittagong-4331, Bangladesh.

A. M. A. Ahmed
Department of Genetic Engineering, University of Chittagong, Chittagong-4331, Bangladesh.

L. W. Marzan
Department of Genetic Engineering, University of Chittagong, Chittagong-4331, Bangladesh.

M. Al-Forkan
Department of Genetic Engineering, University of Chittagong, Chittagong-4331, Bangladesh.

Abstract/ Executive Summary:

The experiment was conducted to identify the in vitro responsiveness for callus induction along with plant regeneration of deep water rice in the presence of NaCl. In this study, callus induction and plant regeneration responsiveness were tested using different concentrations of 0.1-0.3% (w/v) NaCl which added in MS and LS based media. Marked variation was observed both in calli proliferation and plant regeneration among the six deep water rice cultivars. In the presence of 0.1% (w/v) NaCl, the highest number of callus induction in all genotypes recorded on the media supplemented with 2 mg L^-1 2,4-D + 0.1% (w/v) NaCl. The highest dose of NaCl salt 0.3% (w/v) inhibited callus induction compared to 0.1 and 0.2% NaCl media combinations. Among the cultivars tested, cv. Murabajal produced the highest percentage (39%) of callus whilst cvs. Gheoch (23%) and BR224-2B-2-5 (8%) responded poorly in terms of callus production on the MS based 0.1% NaCl supplemented medium. NaCl had no clear promotive effect on regeneration percentage. In this study, green plant did not regenerate from cv. HA-8. Salinity strongly reduced regenerating capacities of callus obtained from all the cultivars. The mean number of shoots produced per regenerating calli was gradually decreased in all the cultivars when the concentration of NaCl increased in the medium. The apparent tolerance of NaCl in these deep water rice cultivars may be at least partially related to its growth rate since it can dilute the contents of ion in the shoot. The results of the present study showed the decreasing trend in callus proliferation and plant regeneration with the increasing concentrations of NaCl.

Keywords: Embryogenic callus, NaCI, Salt-tolerance, Plant regeneration, Deep-water rice

Location: Tissue Culture Laboratory of Botany Department, University of Chittagong, Chittagong, Bangladesh

Start Date: 00-00-2004

End Date: 00-00-2005

Program Area: Crop-Soil-Water Management

Commodity: Rice, Soil salinity

Objectives:

To identify the in vitro responsiveness for callus induction along with plant regeneration of deep water rice in the presence of NaCl

Methodology:

The experiment was carried out at the Tissue Culture Laboratory of Botany Department, University of Chittagong, Chittagong, Bangladesh during the period from 2004-05. Mature Seed Scutellum (MSS) of selected six cultivars namely; HA-1, HA-2, HA-8, Murabajal, Gheoch and BR224-2B-2-5 were used in the present experiments. At first seeds were dehusked and seeds were then washed thoroughly in the running tap water. The floating dehulled seeds were discarded. Then the seeds were taken in to a sterile tube and immersed into 0.2% (w/v) HgCl₂ solution for 15 min. Seeds were then rinsed 4-5 times with sterile deionished water. Then seeds were placed on Murashige and Skoog (1962, MS) and Linsmier and Skoog (1965, LS) based NaCl supplemented callus induction media. MS and LS basal media were supplemented with 2 mg L^-1 2,4-D and 0.1-0.3% (w/v) NaCl, 30 g L^-1 sucrose and 0.8% (w/v) agar. Media were sterilized in an autoclave at a temperature of 121°C for 20 min at 1.16 kg cm^-1 pressure. After 5-6 days of inoculation, seeds of six cvs. germinated on these media. A mass of callus was formed at the scutellar region after 20 days. After 20 days, calli were further inoculated on the same fresh media. After two sub-cultures, the calli showed variation in color and texture. When the texture became compact and color became brownish, the calli were then transferred to MS and LS based regeneration media which were supplemented with only 2 mg L^-1 BAP and 2 mg L^-1 BAP + 0.1% NaCl, semi solidified with 1% (w/v) agar and incubated calli were kept in the dark for 10 days to produce shoots. After that, calli were then sub-cultured on the same medium, semi solidified with 0.8% (w/v) agar and incubated to the light condition. When regenerated shoots grew to about (7-8 cm) in height, they were separated aseptically from each other and transferred to freshly prepared rooting medium supplemented with MS + 2 mg L^-1 BAP + 1.5 mg L^-1 NAA + 0.1% NaCl to induce roots. Finally, well-developed plant lets with sufficient root system were successfully transferred to potted soil along with seed derived control plants.

Source of Information: Asian Journal of Plant Sciences, 6(1): 36-41, 2007, ISSN 1682-3974.

Article Link (Online Journal): DOI: 10.3923/ajps.2007.36.41; URL: http://scialert.net/abstract/?doi=ajps.2007.36.41

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

From these findings it is concluded that the frequency of callus formation and efficient plant regeneration are influenced by the composition of cultured media in the presence of NaCl and also regenerating capacity of embryogenic callus influenced by the genotype. The response of rice cultivars to in vitro tissue culture is under complete genetic control and that separate groups of genes are involved in the control of callus induction, callus growth and plant regeneration.

Knowledge Management: Journal