H.I. Khan
Ex-Student, Agrotechnology Discipline, Khulna University, Khulna
T. Pervin
Ex-Student, Agrotechnology Discipline, Khulna University, Khulna
D. Biswas
Student, Agrotechnology Discipline, Khulna University, Khulna
M.R.K. Bhuiyan
Ex-Student, Agrotechnology Discipline, Khulna University, Khulna
Associate Professor, Agrotechnology Discipline, Khulna University, Khulna
Rice, Seed culture, Spirulina, 2,4-D
Plant Tissue Culture Laboratory of Agrotechnology Discipline, Khulna University, Khulna
Resource Development and Management
The experiment was conducted in the Plant Tissue Culture Laboratory of Agrotechnology Discipline, Khulna University, Khulna, during the period of August, 2006 to January, 2007.
Healthy rice seeds from the variety Kalizira were selected and manually dehusked. They were washed with tap water and then transferred to the laminar airflow cabinet. Then the seeds were surface sterilized by dipping in 70% ethanol for one minute, followed by rinsing in 0.2% HgC12 solution for 15 minutes. The materials were then washed 3 times with autoclaved double distilled water to remove the trace of HgC12. Sterilized seeds were cultured directly onto callus-inducing media with the help of sterilized forceps into the conical flask containing 25 ml of nutrient medium. The cultures were kept in the dark condition for one month at 25±1° C. After that the cultures were transferred under 16 hour photoperiod at 3000 Lux.
Three different media were used for callus induction viz. i) full dose of N6 salts (control), ii) 50% N6 salts +1.6 g rt Spirulina and iii) N6 macro salts + 1.6 g 1-1 Spirulina. Callus inducing media were supplemented with different concentrations of 2, 4 D -1.5, 2.0, 2.5, and 3.0 mg l-1.
When calli attained inconvenient size, after 8 weeks of inoculation of seeds, they were removed and were subcultured on freshly prepared MS medium, containing 3 mg 1-1kinetin for plant regeneration. The culture vessels containing calli were placed under fluorescent light in a growth room with controlled temperature (25±1°C) under a 16-hour photoperiod with a light intensity of 3000 Lux.
After one month of inoculation of rice seeds, callus induction frequency was calculated. Data on morphogenesis were recorded after one month of transference of calli onto the regeneration medium.
The experiment was laid out in the laboratory following Completely Randomized Design (CRD) with three replications. Two factors: media (A) and concentrations of 2, 4-D (B) were considered in this study to investigate their effect on callus induction. For plant regeneration only the effect of 2, 4 D was considered. The recorded data were analyzed for ANOVA with the help of a statistical package program (SPSS-8.0) on a computer. The mean differences were compared by LSD and Duncan's Multiple Range Test (Gomez and Gomez, 1984).
South Asian J. Agric., 2007, 2(1&2):7-10 ISSN 1991-0037
Journal