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The Influence of Carbohydrate as Energy Source in Medium on Germination and Seedlings Growth of Spathoglottis Plicata Orchid in Vitro

M. Obaidul Islam
Professor
Department of Crop BotanyBangladesh Agricultural University, Mymensingh-2202, Bangladesh

S. M. Mahfoozul Muzeer
Department of Crop BotanyBangladesh Agricultural University, Mymensingh-2202, Bangladesh

A. K. M. Masuduzzaman
Department of Crop BotanyBangladesh Agricultural University, Mymensingh-2202, Bangladesh

K. Sharif Mahmud
Horticulturist, Dipta Orchid Ltd., Fulbaria, Mymensingh

Abstract/ Executive Summary:

An experiment was conducted in Tissue Culture Laboratory of the Department of Crop Botany BAU from August 2008 to May 2009 to investigate the effects of carbohydrate supplemented as energy source in medium on seeds germination and seedlings growth of Spathoglottis plicata orchid in vitro. The Hyponex medium of 20 ml and 30 ml with pH 5.8 at 25°±1°C was taken in 100 ml conical flask for seeds germination and seedlings growth, respectively. The culture was maintained at 25⁰C room temperature under 16 hours lighting of 30 µ mol m^-2 s^-1 provided by florescent tubes. Among carbohydrates sucrose at 30 g L^-1 was the best as energy source for seeds germination and seedlings growth in vitro.

Keywords: Carbohydrate, Seed germination, Seedling growth, Orchid, In vitro

Location: BAU, Mymensingh

Start Date: 00-08-2008

End Date: 00-05-2009

Program Area: Variety and Species

Commodity: Cactus

Objectives:

To investigate the effects of carbohydrate supplemented as energy source in medium on seeds germination and seedlings growth of Spathoglottis plicata orchid in vitro.

Methodology:

Plant materials: Mature but indehiscent pod of Spathoglottis plicata collected from plants grown in BAU campus were used for seeds. Capsules contained numerous powders like seeds were preserved in normal freezer at 4 °C. Germination test was done within 2-3 weeks after collection. Culture method and treatments: A clean indehiscent pod was sterilized by dipping into 0.1% HgCl₂ with 2-3 drop tween-20 for 15 minutes inside a clean bench. The pod was then washed 5 times with double distilled water. Dissecting pod by a sharp scalpel seeds were suspended in 100 ml sterilized distilled water and agitated gently by a glass rod. Few drops of suspension were taken on slide and number of seeds per drop was counted under microscope. Five drops suspensions were dispensed uniformly on 30 ml culture medium in 100 ml flask. The pH of the medium was 5.8. Germination percent was recorded after 7, 8, 9 and 10 weeks culture. Protocorm like body (PLB) development was considered as germination because it is an essential and first step in germination process of orchid seeds. Seedlings grown for 7 weeks more in same culture flask (total 17 weeks from dispensing) were subcultured on same fresh medium to investigate their growth in vitro. Each flask contained 30 ml medium and 10 seedlings of similar size at transplantation. The culture temperature was maintained 25°±1°C in a growth room with 16 hours photoperiod of 30 µ mol m^-2s^-1 provided with fluorescent tubes. Five different concentrations of sucrose (10, 20, 30, 40 and 50 gL^-1) as energy source were supplemented into BM and standardized. Furthermore, maltose and sorbitol of equivalent mol. weight were compared to sucrose 20. Each treatment was replicated 5 times. Complete randomized design (CRD) was followed in the experiment. Preparation of Hyponex medium: To prepare 1 litre medium, 3 g Hyponex medium (The Kyoto solution) powder and 10-50 g sucrose (following treatments) were taken separately in 1 liter erlenmeyer flask. Adding distilled water (DW) sucrose was dissolved and volume was made about 990 ml and pH was adjusted to 5.8 at 25°C temperature. Pouring DW volume was made 1 L and then 3g gelrite (Merck and Co., Inc.) was added to the solution as solidifier. By heating in water bath with stirring gelrite dissolved. The medium was then dispensed at 20 ml in each 100 ml conical flasks, capped with aluminum foil and finally autoclaved. Data recording and analysis: Numbers of PLB initiated on medium were counted and % germination calculated. Similarly data on seedlings growth were collected after 3, 6 and 9 weeks culture and analysed by computer programe developed by Russel (1986) and means were compared to least significant differences (LSD) test for interpretation of results.

Source of Information: Bangladesh J. Seed Sci. Technol. 13(1&2): 83–88 (2009), ISSN 1029-8800

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In the present experiment, sucrose was found as an essential factor for germination of seeds of Spathoglottis plicata. During germination sugar metabolizes into glucose as energy in germinating embryo. It is not clear that whether germinating embryo converts sucrose into glucose inside cells or outside of cells during culture. Sucrose cleavage is vital to multicellular plants, not only for the allocation of crucial carbon resources but also for the initiation of hexose-based sugar signals in importing structures. Among carbohydrates sucrose at 30 g L^-1 was the best as energy source for seeds germination and seedlings growth in vitro.

Knowledge Management: Journal