Agricultural Research Management Information System

Home
Research Summary
Search
About Us
- ARMIS
- Brochure
Contact Us
Report
User Request
Data Input
Help
- Operation Manual
  - PDF
  - Video
- Program Area & Commodity
We have reached 37600 number of research entries at this moment.

- Logout

Research Detail

Home
Research
Detail

Ontogeny of Embryogenic Aggregates in Suspension Culture of Diploid Watermelon [citrullus Lanatus (thunb.)]

Rubaiyat Sharmin Sultana
Department of Botany, University of Rajshahi, Rajshahi 6205, Bangladesh.

Md. Mahabubur Rahman
Research Institute of Sustainable Humanosphere, Kyoto University, Uji, Kyoto 611 - 0011, Japan.

Abstract/ Executive Summary:

Cell suspension culture from leaf - derived callus of diploid watermelon [ Citrullus lanatus (Thunb.)] was established. The callus obtained on agar - gelled Murashige and Skoog (MS) medium containing 2.5 mgl^-1 2,4 - dichlorophenoxyacetic acid (2,4 - D) was used for suspension culture. The growth of cell in the suspension culture was the highest in liquid MS medium containing same concentration ( 2.5 mgl^-1 ) of 2,4 -D as used for callus induction. Cells in suspension culture underwent division as result both free cells and cell aggregates were formed. In first 15 days, profuse number of early proembryogenic structures (two -, three - and four - celled) was observed while after more 15 days of culture, multi - celled proembryogenic structure was greater in suspension. The initial amount of cell in suspension culture was effective for cell proliferation and cell aggregation. As initial amount of cells, 4 ml sedimented cell volumes (SCV) could proliferate and aggregate at a highest level after 30 days of culture. Three typical phages (lag, exponential and stationary phases) in S - shaped growth curve were found in the batch culture. Among the cell growth phages, duration of exponential phage is important for the high production of cell and biosynthesis of cell compounds.

Keywords: Callus, Cell aggregate, Cell suspension, Proembryogenic structures, Watermelon

Location: Department of Botany, University of Rajshahi, Rajshahi

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Water melon

Objectives:

To established cell suspension culture and observed cell morphology and ontogeny of cell aggregates of watermelon.

Methodology:

Seeds of watermelon were collected from local market and used for further investigation.As a basal medium, MS medium was used for all experiments studied here. The pH of media was adjusted to 5.7±0.1 and 0.8% (w/v) agar was added to medium solidification before autoclave at 121ºC for 20 min under 1.1kg cm^-2 pressure. The medium without plant growth regulators (PGRs) used as control for all experiments. The cultures were maintained in a growth chamber controlled with 27± 1 ºC and a 16 -h photoperiod (35 μmol m ^{- 2} s^-1 ). Callus was achieved followed by our previous protocol on organogenesis of watermelon. The induced callus ( 5 g fresh weight) on MS medium fortified with 2.5 mgl^-1 2,4 - dichlorophenoxyacetic acid (2,4 - D) was transferred to a 300 - ml Erlenmeyer flask containing 50 ml of liquid MS medium fortified with either 0.1 and 2.5 mgl^-1 2,4 - D alone or in combinations with 0.5 mgl^-1 6 - benzylaminopurine (BAP). The flasks were sealed with Aluminum foil, wrapped with parafilm and then they were placed on a rotary shaker (90 rpm). Callus cells were proliferated in suspension cultures for 15 days. The cell growth in each PGR treatment was measured. For measurement of cell growth, the proliferated cells in an Erlenmeyer flask were dispensed in a sterile 1 00 ml measuring cylinders and allowed to sediment for 30 min after that measured total cell amount by sedimented cell volume (SCV) as milliliter (ml). Proliferated cells from initial suspension at different amounts (2, 4, 6 and 8 ml SCV) were transferred in 300 ml Erlenmeyer flasks containing 50 ml of MS liquid medium supplemented with 2.5 mgl^-1 2,4-D. For this experiment, initially established suspension was first filtered several times to avoid cell aggregates and then transferred to fresh medium routinely in a week. After 15 days and 30 days of culture, cell suspension observed under light microscope. Additional cell amount was measured by SCV ml. The number of cell aggregates was also counted after 30 days of culture in all suspension using hemacytometer followed by method that used for cell suspension culture of sweet potato. Initially proliferated cells within 15 days were used to examine cell growth by batch culture. Therefore, 4 ml SCV cells were cultured in 300 ml Erlenmeyer flasks containing 50 ml MS liquid medium fortified with 2.5 mgl^-1 2,4 - D and routinely transferred to a fresh medium at one - week - interval. The cultures were maintained up to 8 weeks. Total amount of cell (SCV ml) per flask measured prior to transfer in each transfer. The number of replications was three for all experiments and experiments were repeated at least thrice . The experiments were conducted with a completely randomized design and the data were analyzed by analysis of variance. To distinguish differences among the mean value of treatments, Tukey’s multiple comparison test using JMP Statistical Discovery Software (SAS Institute, USA) was used, the least significance difference (LSD) test at 5% ( p ≤ 0.05) level was used.

Source of Information: Int. J. Agr. & Agri. R. Vol. 2, No.1, p. 40-46, 2012, ISSN:2223-7054

Article Link (Online Journal): http://www.innspub.net/wp-content/uploads/file/IJAAR-V2No1-p40-46.pdf

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The initial amount of cell in suspension culture was effective for cell proliferation and cell aggregation. As initial amount of cells, 4 ml sedimented cell volumes (SCV) could proliferate and aggregate at a highest level after 30 days of culture. Three typical phages (lag, exponential and stationary phases) in S - shaped growth curve were found in the batch culture. Among the cell growth phages, duration of exponential phage is important for the high production of cell and biosynthesis of cell compounds.

Knowledge Management: Journal