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In Vitro Propagation in Three Varieties of Gerbera (gerbera Jamesonii Bolus.) from Flower Bud and Flower Stalk Explants

Nazma Akter
Plant Breeding and Biotechnology Laboratory, Department of Botany, University of Dhaka, Dhaka

M. I. Hoque
Plant Breeding and Biotechnology Laboratory, Department of Botany, University of Dhaka, Dhaka

R. H. Sarker
Plant Breeding and Biotechnology Laboratory, Department of Botany, University of Dhaka, Dhaka

Abstract/ Executive Summary:

Three types of tender leaves derived explants such as leaf tip, leaf with mid-rib and leaf blade segments as well as flower buds and flower stalks obtained from three selected varieties of gerbera (Gerbera jamesonii Bolus.) cultivated in Bangladesh were exploited for callus induction and in vitro regeneration of plantlets. Among the explants flower bud and flower stalk were suitable and superior for callus induction and subsequent regeneration of in vitro shoots when cultured on MS supplemented with 5.0 mg/l BAP and 1.0 mg/l NAA. However, highest number of multiple shoots were obtained when the flower bud derived callus was subcultured on MS supplemented with 2.0 mg/l BAP. Variety with red petal showed the best response in producing multiple shoots among the three varieties. Root induction at the base of the in vitro regenerated shoots was tried using full and half the strengths of MS containing different concentrations of IBA. Best root induction was observed on half the strength of MS supplemented with 0.2 mg/l IBA. Following adequate acclimatization the regenerated plantlets were successfully transferred to soil where they grew to maturity and produced flowers.

Keywords: Propagation, Gerbera, Flower bud, Flower stalk

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Gerbera

Objectives:

The present investigation was undertaken with a view to developing an in vitro plant regeneration protocol for large scale propagation of selected Gerbera varieties cultivated in Bangladesh using suitable explants

Methodology:

Plants of three varieties of gerbera (Gerbera jamesonii Bolus.) having red, yellow and white coloured petals collected from BRAC Research and Development Centre, Joydebpur, Gajipur and were used for this study. Tender leaves and 2.5 - 4.0 cm long flower buds (9 - 11 days old) along with flower stalks were collected from the field grown materials. Special treatments were applied to reduce the level of contamination since field grown materials were used for this investigation. Leaves and flower buds with stalks were first washed under running tap water for 30 min. Then the explants were washed with detergent and then the detergent was washed out completely. The explants were then immersed in 20% Savlon (v/v), an antiseptic disinfectant containing chlorhexidine gluconate and cetrimide (Novartis Consumer Health UK Ltd.) for 2 min followed by washing with distilled water. After that the explants were treated with a fungicide called bendagime (1.0 gm/500 ml) for 5 min followed by several washing with distilled water. The explants were then deepened in 70% alcohol for 30 seconds followed by washing three times with distilled water. The final surface sterilization of the explants was done with 0.1% HgCl₂ solution for 6 - 7 min inside the laminar flow cabinet. During this operation, the flask was agitated frequently and finally the explants were washed five times with sterilized distilled water. For inoculation the surface sterilized leaf explants were cut into three pieces, namely leaf tip, leaf with mid-rib and leaf blades. One - two cm long segments of these leaves derived explants were cultured for callus induction and shoot multiplication. On the other hand the surface sterilized flower buds were dissected into 8 - 10 pieces for using them as explants. About 1 - 2 cm long segments of petiole and flower stalks were also used as explants for callus induction and in vitro regeneration. MS supplemented with different concentrations and combinations of BAP (1.0 - 6.0 mg/l), NAA (0.5 - 2.0 mg/l) were used for the induction of callus and development of multiple shoots from different explants of three varieties. All media contained 3% sucrose and 0.8% agar with pH 5.8, adjusted before autoclaving. For rooting 3 - 4 cm long regenerated shoots were excised and cultured on freshly prepared rooting medium containing full and half strength of MS with different combinations and concentrations of IBA and NAA. All cultures were maintained less than 16 hrs photoperiod at 25 ± 2°C. Following the development of sufficient roots, plantlets were transferred to small plastic pots containing sterilized soil. These plantlets were acclimated and then transferred to the field and raised there till their maturity to flowering.

Source of Information: Plant Tissue Cult. & Biotech. 22(2): 143-152, 2012 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Induction of healthy root system from the regenerated shoots is an essential part for successful development of plantlets. During the present study it was noticed that several roots developed spontaneously from the in vitro grown shoots but the spontaneously developed roots were found to be inadequate for transplantation of the in vitro grown shoots to the soil. Therefore, separate root induction was necessary. For root induction, regenerated shoots were cultured on half and full strength of MS supplemented with IBA, IAA and NAA. It was observed that half strength of MS with 0.2 mg/l IBA showed 95 - 100% for root induction in the red, yellow and white varieties. When NAA was used at 0.1 and 0.2 mg/l with half the strength of MS, only callus was induced. Efforts were made to establish plantlets in pots with proper root systems. Plants with sufficient roots were transplanted to soil ) and their survival rate was 100% in the field condition. These tissue culture derived plants flowered very much identical to that of the control plants. All the three Gerbera varieties used in this study showed almost identical responses towards in vitro regeneration. The overall responses of the red petal variety were found to be relatively better compared to the other two varieties of local gerbera. Through this investigation it has been possible to develop an efficient and reproducible in vitro mass propagation system from different explants of gerbera. Based on the results it may be concluded that the in vitro mass propagation protocol developed in this investigation could profitably be explored for the commercial cultivation of gerbera. The present protocall of propagation may help in obtaining adequate number of plantlets for cultivation in fulfilling the requirements of the local market of gerbera.

Knowledge Management: Journal