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Effect of Growth Regulators on in Vitro Multiplication of Dianthus (dianthus Chinensis)

F. Akter
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh.

S.M.A. Awal*
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh.

S.M.M. Rahman
Biotechnology and Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh.

Abstract/ Executive Summary:

The experiment was conducted to clarify the effective concentrations of cytokinin and auxin on shoots and roots formation of Dianthus chinensis. Shoot tip from in vitro germinated seedlings was inoculated onto modified MS medium supplemented with the various combinations of BA and NAA. Among them BA 10.0 mg1-1+ NAA 0.5 mgl-1 gave the highest number of multiple shoots (10.75). The highest number of usable shoots (7.65) was obtained with the same concentration. The highest shoot length (6.13) was observed at BA 0.5 mgri + NAA 0.025 mg1-1. The in vitro grown shoots were tested for rooting in both semi-solid and liquid MS medium supplemented with similar concentration of NAA, IBA and IAA where IAA produced the maximum number of roots in both of the semi-solid (10.65 at 0.05 mgl-1) and liquid media (19.00 at 0.5 mgi'). Maximum root length (4.6 cm) was also obtained from 0.5 mgri IAA in liquid media, IBA at 0.05 mei gave maximum root length (3.93 cm) in semi-solid media.

Keywords: Dianthus, Auxin, Cytokinin, Shoot and root

Location: Plant Biotechnology Laboratory of Biotechnology and Genetic Engineering Discipline, Khulna University, Bangladesh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Growth regulator, Flower

Objectives:

Micropropagation has already a proven record of increased yield and vigor through high health programmes in floriculture, and a big role to play in the rapid introduction of new varieties of ornamentals into the market. It is clear that if the correct strategies for the use of micropropagation are developed and used in the appropriate circumstancs, the techonology will have a major effect in the next decades on both the nature of the crops that are grown internationally and also on the whole structure of horticulture and agronomy worldwide (Purohit, 2002). Hense, considering the above staments, the present study was conducted to verify the suitable concentration and combination of cytokinin (BA) and auxin (NAA) for shoot proliferation and to select the optimum concentration of auxin for effective rooting after light treatment for root induction.

Methodology:

The experiment was conducted at Plant Biotechnology Laboratory of Biotechnology and Genetic Engineering Discipline, Khulna University, Bangladesh. Seeds of D. chinensis were collected from elite ornamental flower growers for the study. Seeds were first washed under running tape water in the laboratory to reduce dust and emersed in 0.1% HgC12 solution with shaking for 5 min aseptically. The seeds were then rinsed 3-4 times (each time 5 min) with sterilized distilled water to remove the trace of the sterilant. The seeds were then germinated aseptically onto MS (Murashige and Skoog, 1962) media without growth regulator(s). Seeds were inoculated into 200 test tubes, each tube containing 15 ml media. For germination test half of the inoculated test tubes were kept under dark condition at 26±2 °C and other half was maintained under light illumination (300 lux) provided from white cool fluorescent tube. After 15 days of inoculation the test tubes were observed and the rate of germination was recorded. The pH of the medium was adjusted between 5.7 and 5.8 using a digital pH meter with the help of 0.1 N NaOH or 0.1 N HCI prior to autoclaving at 121°C for 20 min at 15-psi pressure.

After 4—weeks of germination, shoot tips were collected from in vitro germinated seedlings under aseptic condition and inoculated onto MS medium supplemented with different concentrations and combinations of BA and NAA. Each treatment consisted of 20 tubes for multiplication. The germinated seedlings were aseptically taken out of the culture vessels and cut into pieces of 1 cm. long with scalpel in a laminar flow and inoculated into the medium.

The MS medium supplemented with IAA, IBA and NAA was used for root induction. Semi-solid and liquid media were tested for this study. After four weeks of shoot multiplication, elongated shoots (usable shoots, 1.5- 2.0 cm) were used for root induction. All the cultures at different steps were incubated at 25±1°C under light illumination (3000 lux) provided from white cool fluorescent tubes with photoperiod of 16h light and 10h dark. After 4 weeks of culture rooted plants were taken out from the test tubes and rinsed with tape water to remove the medium. Plantlets were then transferred to pot containing 75% soil and 25% sand. For gradual hardening, pots were covered with perforated polythene sheets to reduce the evaporation for few days and then explised to sunlight. The pots were irrigated whenever necessary. Some plantlets were died due to fungal and bacterial contamination. However 60% plants survived and hardened properly and transfer to the soil. The data pertaining to shooting and rooting per culture were subjected to statistical analysis and were analyzed using ANOVA and Dancan's Multiple Range Test (DMRT).

Source of Information: South Asian J. Agric. 2009. 4 (I &2): 128-131 ISSN 1991-0037

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

It can be concluded that Dianthus chinensis explant carry a high potential for shoot formation and rooting on MS culture media. In the studies, a successful attempt has been made to regenerate complete plantlet from excised explant of in vitro germinated seedlings of Dianthus chinensis by manipulation of nutrient media and hormonal combination.

Knowledge Management: Journal