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Effect of Colchicine on Fruit and Seed Development in Regenerated Teasle Gourd Plant

S. S. U. Ahmed
Ph.D Student.
Department of Crop Botany. Bangladesh Agricultural University. Mymensingh-2202. Bangladesh

M. A. Sarnad
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, BAU Campus, Mymensingh-2202. Bangladesh

M. A. Karim
Department of Crop Botany, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. S. Haque
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202. Bangladesh

Abstract/ Executive Summary:

Tissue culture technique was applied for micropropagation of teasle gourd (Momordica dioica Roxb.) to achieve seedless fruit applying colchicine on callus. The study included the number of fruits per plant and its size, protein content of flesh and number of seeds fruit^-1. Number of fruits per plant from natural, regenerated and colchicine treated regenerated plants were 8.2, 7.4 and 7.0, respectively. Treated regenerated plants produced medium sized (6.6 cm) fruit which was larger than natural (6.1 cm) but smaller than regenerated one (9.1 cm). The seed number of colchicine treated fruit drastically reduced to 14.6, whereas natural and regenerated fruit had 33.2 and 26.6 seeds, respectively. The protein content of flesh varied in natural, regenerated and treated regenerated fruits. Edible flesh of 17 days old regenerated fruit had the maximum 7.08% protein, whereas the treated regenerated flesh resulted 4.62% protein followed by 5.54% in natural fruit flesh. Thirty days old fruit flesh had similar trend of protein content.

Keywords: Colchicine, Micropropagation, Seedless fruit, Teasle gourd

Location: Agricultural University. Mymensingh-2202. Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Teasle Gourd

Objectives:

i) To induce variations among the germplasms using chemical mutagensc.

ii) To screen the desired plant variants for quality fruits (seedless or reduced seeded fruit) and high yielding plant types.

Methodology:

Fast growing callus was obtained from cotyledon of immature fruit from the teasle gourd. Among hormonal supplements, 1 mgL^-1 BA was useful, The detailed procedure for explant sterilization, establishment and maintenance of callus and regeneration protocol were described earlier. All media were adjusted to pH 5.8 prior to sterilization. Cultures were kept at 24 ± 2°C. For colchicine experiment, colchicine was dissolved in water for the preparation of 0.05% solution. Colchicine treatment was given for 1, 2, 4 and 8 hours. The flasks containing 20 callus pieces flask^-1 soaked by the liquid medium. A few calli showed necrosis and early death. After treatment for a varying length of time, surviving calli were washed several times with sterilized distilled water and transferred to solid regeneration medium that contained 1 mgL^-1BA. For another type of treatment, colchicine was mixed with MS medium at 10, 20, 30 and 40 mgL^-1 for 3, 6 and 9 days. After 3, 6 and 9 days culturing on colchicine supplemented media, the calli were subcultured on MS medium with 1 mgL^-1 BA. Sterilization of colchicine was made by using filter (Pore size 0.22 mesh). Fruits from natural, regenerated and treated regenerated plants were taken for measurement of protein content. One gram of sample (oven dried seed and flesh of fruit) was weighed accurately and taken in digestion flask (Kjeldahl flask). Three to 4 gm of digestion mixture (CuSO₄: K₂SO₄ = 1 :5) were added to it. Twenty milliliter of concentrated H₂S0₄ was added into the flask. The flask was placed on digestion chamber and boiled until the solution became clear. The flask was cooled slowly and 150 ml distilled water was added to it. Some glass beads and zinc granules were added to avoid bumping. Hundred 1 ml of 40% NaOH solution was poured into the flask very slowly holding the flask about 45° angle and immediately the flask was connected to a distillation set. The liberated ammonia was collected in a conical flask which was previously placed under the condenser jet containing 25 ml of 2% boric acid solution and 2-3 drops of mixed indicator (methyl red and methyl blue). At least 125 ml distillate was collected into the conical flask. Frequent bumping or the flask indicated the completion of the distillation. Distillation was titrated with 0.2N HCl. The Calculation is shown as below: % Protein= % Nx 5.5, % N={ (T_S-T_B ) X normality of acid X 0.014/ Wt. of sample (in gm)} X100. Here, T_S= Titre value of the sample, T_B= Titre value of the Blank, 0.014=Milliequivalent of nitrogen and 5.5 is a conversion factor. Results were expressed in grams of protein per 100 grams of experimental sample.

Source of Information: Bangladesh Journal of Nuclear Agriculture. Vol: 30, pp: 37-42, ISSN 0258-7130, 2014

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The seeds of natural fruit had the maximum (16.63%) protein content which was similar to the treated regenerated fruit (16.02%), whereas the regenerated seed had the minimum (12.94%) protein content. The highest protein accumulation was found in fruits of regenerated plant and the reduced number of seeds were found by the application of colchicine.

Knowledge Management: Journal