Agricultural Research Management Information System

Home
Research Summary
Search
About Us
- ARMIS
- Brochure
Contact Us
Report
User Request
Data Input
Help
- Operation Manual
  - PDF
  - Video
- Program Area & Commodity
We have reached 37600 number of research entries at this moment.

- Logout

Research Detail

Home
Research
Detail

In Vitro Regeneration and Screening for Salt Tolerance in Rice (oryza Sativa L.)

K.M.A. Zinnah
Dept. of Genetic Engineering and Biotechnology, School of life sciences, Shahjalal University of Sci. and Tech., Sylhet-3114, BANGLADESH

Nayem Zobayer
Dept. of Genetic Engineering and Biotechnology, School of life sciences, Shahjalal University of Sci. and Tech., Sylhet-3114, BANGLADESH

Saif U. Sikdar
Dept. of Genetic Engineering and Biotechnology, School of life sciences, Shahjalal University of Sci. and Tech., Sylhet-3114, BANGLADESH

Lutfun Nahar Liza
Dept. of Genetic Engineering and Biotechnology, School of life sciences, Shahjalal University of Sci. and Tech., Sylhet-3114, BANGLADESH

Md. Al Nayem Chowdhury
Dept. of Genetic Engineering and Biotechnology, School of life sciences, Shahjalal University of Sci. and Tech., Sylhet-3114, BANGLADESH

M. Ashrafuzzaman
Dept. of Genetic Engineering and Biotechnology, School of life sciences, Shahjalal University of Sci. and Tech., Sylhet-3114, BANGLADESH

Abstract/ Executive Summary:

The study was conducted to obtain salt tolerant genotype of BRRI Dhan 38 and Chini Kanai (local variety) rice varieties through somaclonal variation. Different concentration and combinations of growth regulators were supplemented to MS medium to observe the callus induction and plantlet regeneration ability of mature rice seeds. On the other hand, the calli were transferred to the best regeneration medium at different concentrations of NaCl to check the inherent capacity of calli to regenerate on medium under salt stress condition. Maximum percentage of callus induction was observed in MS medium supplemented with 5 mg/l 2,4-D for BRRI Dhan 38 and 3 mg/l for Chini Kanai. Calli derived from the different concentrations of 2, 4-D were cultured on MS medium supplemented with 1 mg/l NAA, 2 mg/L BA and various concentration of Kinetin for plantlet regeneration. It was observed that MS media supplemented with 2 mg/l of kinetin in combination with 1 mg/l NAA and 2 mg/l BA produced highest percentage of callus for BRRI Dhan 38 (80%) and Chini Kanai (60%) respectively. Plant regeneration of BRRI dhan 38 was 80% at 0 mM NaCl, but decreased to 20% at 100 mM NaCl. There was 0% plant regeneration at 150 mM NaCl for BRRI 38 and Chini Kanai respectively. In Chini Kanai plant regeneration on the no-stress medium was 60%. At 150 mM it decreased to 20% and there was no regeneration at 200 mM NaCl. It indicates that Chini Kanai is more salt tolerant then BRRI Dhan 38.

Keywords: Oryza sativa, Plant growth regulators, Salt tolerency.

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Rice

Objectives:

In recent years tissue culture techniques are being used as a useful tool to elucidate the mechanism involved in salt tolerance by using in vitro selected salt tolerant cell lines. Besides, these lines have been used to regenerate salt tolerant plants.

Methodology:

This experiment was conducted in the plant genetic engineering laboratory of the Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology (SUST), Sylhet, Bangladesh. During this field grown seeds of aromatic rice (Oryzasativa L.) variety were used for callus induction, plant regeneration and salt screening. Explant source: Mature seeds were used as explants in this experiment. The rice variety (Oryza sativa) BRRI 38 and CHINI KANAI were collected from the Bangladesh Rice Research Institute (BRRI) and Paikgasa, Khulna respectively. Explant Sterilization: The mature embryo was sterilized by following the standard explants sterilization protocol, previously described by different researchers. The explants were thoroughly washed with distilled water for three times. Then, the explants were washed with 70% ethanol (for 2 to 3 min). In the laminar air flow cabinet, the explants were treated with 0.1% HgCl2 with the addition of few drops of Tween-20 for inner surface sterilization (for 4 to 6 min). Finally, the explants were washed with sterile distilled water for several times to remove all the sterilizing agents. Callus induction media: The basal medium MS was used for callus induction. The proposed medium was supplemented with concentration of growth hormone 2,4-D (0, 1, 2. 3,4,5 and 6 mg/L). The pH of the media was adjusted to 5.8 ± 2 before autoclaving. After inoculation, the surface sterilized seeds of two rice varieties were transferred and maintained in an environmentally controlled growth room for 3 weeks for callus induction and growth. The cultures were positioned away from continuous light provided by general electric white florescent tubes. Temperature was maintained at 25 ± 3 ⁰C thought the growth period. Callus induction frequency for two varieties was recorded 7–9 days after inoculation. Callus quality was recorded at 2-3 weeks after inoculation in two rice varieties for all treatments. The callus contained both embryogenic (white to light yellow in color, compact and friable) as well as non- embryogenic (mucilagenious and smooth) parts. Subculture was carried out once in every two weeks with transfer of only the vigorously growing portions of calli. The embryo calli were induced from the scutellar tissues of mature seeds, excised and used in later experiments for regeneration after subculturing (to exploit the full potential of cell growth) for 3 weeks on the same media used for callus induction. Experiments were replicated three times and twelve test tubes with twelve seeds were used per replication for each genotype. All the calli originated from a single seed was considered as one. Regeneration Media: For plant regeneration, the embryogenic part of calli was cut into small pieces by removing non embryogenic part. Calli were then inoculated on regeneration media with different combination and concentrations of hormones (Tab.2.8.). The MS basal medium was supplemented with 3% sucrose and kinetin (1, 2, 3 and 4 mg/L) while maintaining 1 mg/L of NAA and 2 mg/l of BA as constant. The pH of media was adjusted to 5.8 ± 2 before autoclaving. Sixteen calli were inoculated on the regeneration media and the culture was performed at 25 ± 3ºC under a cycle of 16 hours light/8 hours dark for 4 weeks, after which the frequencies of plant regeneration were calculated., based on the appearance of shoots. Media for screening salt tolerance: Four-week-old callus was divided into pieces of 100 mg. These pieces were transferred onto the same medium those were used for plant regeneration, supplemented with different NaCl concentrations, such as, 0, 50, 100, 150 and 200 mM, for salt stress responses. At the end of the four-week period, the callus was taken for growth analysis.

Source of Information: International Research Journal of Biological Sciences Vol. 2(11), 29-36, November (2013) ISSN 2278-3202

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In vitro tissue culture could be an important means of improving crop tolerance and yield through genetic transformation as well as by induced somaclonal variation. Therefore it is important to devise an efficient protocol of callus prolife ration to start in vitro selection for salt stress tolerance, and to extand opportunities for genetic manipulation of rice through tissue culture, including trying various explants and media.

Knowledge Management: Journal