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Potency of Sucrose Concentration, Ph and Strength of Ms Media on Lettuce Callus Proliferation

A. N. M. Erfan
MS student
Department of Horticulture, Patuakhali Science and Technology University, Patuakhali-8602, Bangladesh.

Abstract/ Executive Summary:

The fresh and dry weight of calli varied significantly on different MS media, different doses of sucrose and different pH levels. The maximum fresh weight (1.72 g) and dry weight (0.17 g) of calli were obtained in full strength MS medium at 45 DAC. On the other hand, the minimum fresh (0.92 g) and dry (0.10 g) weight of calli were produced by the quarter strength MS medium at 45 DAC. Full strength MS medium supplemented with 30 g/L and 20 g /L sucrose provided the highest (1.92 g) and the lowest (1.08 g) fresh weight of calli at 45 DAC, respectively. The maximum (0.24 g) and the minimum (0.10 g) dry weight were gained by the 30 g/L and 20 g/L sucrose levels at 45 DAC, respectively. Full strength MS medium with pH 5.8 gave the utmost fresh (1.52 g) and dry (0.17 g) weight of callus whereas pH 6.2 gave the least fresh (0.71) and dry (0.08 g) weight of callus at 45 DAC, respectively. Full strength MS medium supplemented with 30 g/L sucrose having pH 5.8 provided the best results for callus proliferation.

Keywords: In vitro regeneration, Lettuce, MS media, pH level, Sucrose concentration.

Location: PSTU

Start Date: 01-10-2012

End Date: 31-03-2013

Program Area: Development of Host and Medicinal Plants

Commodity: Lettuce

Objectives:

To develop a protocol for micro propagation of lettuce for high frequency plantlet regeneration, and to determine the best doses of sucrose concentration, pH level and MS medium for callus proliferation.

Methodology:

The study was conducted at the Plant Biotechnology Lab, Department of Horticulture, Patuakhali Science and Technology University, during October 2012 to March 2013. The experiment was laid out in CRD with 4 replications. Newly emergence (one month aged) leaves of the winter lettuce cv. “Grand Rapids” was used as plant material. Different concentrations of sugar (10 g/L, 20 g/L, 30 g/L and 40 g/L ), levels of pH (5.6, 5.8, 6.0 and 6.2), and different strength of MS medium (full, half and quarter strength) on callus proliferation. Separate stock solutions for macronutrients, micronutrients, iron source, vitamins/organics and growth regulators, etc. were prepared prior to medium preparation and stored in a refrigerator at 4°C for subsequent use. The surface sterilized explants were then rinsed 4-5 times with sterile distilled water inside the clean bench to remove all traces of HgCl₂. Leaves were cut by a sharp knife and 1 or 0.5 mm segments were used as explants. The excised explants were then inoculated into each culture bottle containing MS medium with various concentration and combinations of hormonal supplements for in vitro callus initiation. The culture vessels with inoculated explants were incubated in both dark and light in a temperature controlled growth room (25 ± 1°C) under 16 hours photoperiod with a light intensity 1500 lux and relative humidity 60-70%. Plantlets of 45 day old were used as source explants for getting contamination free plantlets. The collected data were fresh weight of callus and dry weight of callus. The Analysis of Variance was performed and means were compared by Least Significant Difference (LSD) test for interpretation of results. The significance of the difference between the pair of means was evaluated at 1% level of significance using MSTAT-C computer package programs.

Source of Information: J. Patuakhali Sci. and Tech. Univ. 2014, 5(1): 11-17

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present experiment, full strength MS medium supplemented with 30 g/L sucrose having pH 5.8 provided the best results for callus proliferation. Further experiment can be conducted by using different concentration and combination of growth regulators, sucrose doses and pH levels in different strength of MS media to find out suitable combination.

Knowledge Management: Journal