Md. Riyadh Arefin
Scientific Officer
Botany Division, Bangladesh Tea Research Institute, Srimangal-3210, Moulvibazar, Bangladesh.
Dr. Mohammad Mehfuz Hasan Saikat
Professor
Department of Genetics and Plant Breeding, Bangabandhu Sheikh Mujibur Rahman Agricultural University (BSMRAU), Gazipur, Bangladesh.
Mr. Umakanta Sarker
Professor
Department of Genetics and Plant Breeding, Bangabandhu Sheikh Mujibur Rahman Agricultural University (BSMRAU), Gazipur, Bangladesh.
Dr. M. Abdul Karim
Professor
Department of Agronomy, Bangabandhu Sheikh Mujibur Rahman Agricultural University (BSMRAU), Gazipur, Bangladesh.
Md. Sadiqur Rahman
Scientific Officer
Seed Technology Discipline, Bangladesh Agricultural Research Institute (BARI), Akbarpur, Moulvibazar, Bangladesh.
Rice, Chloride, Sulphate, In vitro, Regeneration
Advanced Plant Breeding Laboratory, Department of Genetics and Plant Breeding, Bangabandhu Sheikh Mujibur Rahman Agricultural University (BSMRAU), Salna, Gazipur-1706, Bangladesh.
Crop-Soil-Water Management
The following rice genotypes including a salt tolerant variety (BINA 8) were used as experimental materials in the present investigation.
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Sl. No.
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Rice genotypes
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Sl. No.
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Rice genotypes
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1.
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BR10
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6.
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BINA 8
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2.
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BR11
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7.
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Uknimadhu
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3.
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BRRI dhan49
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8.
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Binaphul
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4.
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BRRI dhan51
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9.
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Gopalbhog
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5.
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BRRI dhan52
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10.
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Straw
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Salt concentrations and media preparation
The healthy, disease free and dehusked seeds were used as explants for the study. MS (Murashige and Skoog, 1962) medium supplemented with two salts (NaCl and Na2SO4) was used for callus induction and the callus pieces at appropriate size were transferred to MS regeneration medium.
For callus induction, the MS (Murashige and Skoog, 1962) medium supplemented with 2, 4-D (2mg/l), sucrose (30g/l), agar (10g/l) and six levels of salt concentrations was taken for each salt. For plant regeneration the MS (Murashige and Skoog, 1962) medium supplemented with sucrose (30g/l), agar (l0g/1), NAA (l mg/L), BAP (1 mg/l) and six levels of salt concentration was used for each salt. Six levels of NaCl and Na2S04 salt solution (0, 0.1, 0.2, 0.4, 0.6 and 0.8%) were used in this experiment to observe the effect of salts.
Callus induction and regeneration
The dehusked seeds were sterilized and the seeds were air dried in the Laminar Air Flow cabinet for ten minutes and were transferred on callus induction media. This process was done for each genotype. The culture was inoculated with the treated seeds in the dark at 25 ± 2°C for callus induction. After 3-6 weeks of inoculation, seeds of the responsive varieties started to produce callus. Callus induction frequency was calculated on the basis of the number of seeds producing callus. Some seeds produced more than one callus, but for this calculation, all calli originating from one were considered as one. Calli with a size of at least 2 mm were transferred to regeneration medium (MS basal semisolid medium), and were incubated in a temperature controlled growth room at 25 ± 2°C under 16-hour light photoperiod with a light intensity of about 2000-3000 lux for plant regeneration. Day to day observations was carried out to note the response. Regenerated plants were counted on the basis of the number of callus producing plantlets.
Statistical analysis of data
The data for the characters under study were statistically analyzed wherever applicable. The experiment was conducted in growth room and arranged in Factorial Completely Randomized Design (CRD) with three replications. The Analyses of Variance for different characters were preformatted and means were compared by the Duncan's Multiple Range Test (DMRT).
International Journal of Biosciences | IJB | Vol. 13, No. 1, p. 36-41, 201
ISSN: 2220-6655 (Print), 2222-5234 (Online)
Vol. 13, No. 1, p. 36-41, 2018
Journal