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Development of Protocol for in Vitro Culture and Rapid Propagation of Wedelia Chinensis (osbeek) Merr.

M. M. Rahman
Department of Botany University of Chittagong Chittagong-4331, Bangladesh.

S. K. Bhadra
Department of Botany University of Chittagong Chittagong-4331, Bangladesh.

Abstract/ Executive Summary:

Three different explants of Wedelia chinensis (Osbeek) Merr. namely nodal segment (NS), shoot apex (SA) and leaf segment (LS) were grown on MS basal medium supplemented with different combinations and concentrations of cytokinin (BAP and Kn) and auxin (IAA and NAA). Nodal segment and shoot apex underwent direct organogenesis giving rise to multiple shoot buds and the response was dependent on both PGR combinations and type of explant. The maximum number of multiple shoot buds/explant (7.5±0.27) was developed from nodal segment culturing on MS +3.0 mg/l BAP+0.5 mg/l IAA. But shoot apex explant produced longest shoot (5.12±0.17 cm) when only 2.0 mg/l Kn was used in MS medium. Elongated multiple shoot buds were grown on rooting media. Half strength MS fortified with 2.0 mg/l IBA was found better for induction and proliferation of roots. The in vitro developed complete plantlets were acclimatized in outer environment through successive phases of acclimatization. On an average 80% of the seedlings could be finally established in pots.

Keywords: In vitro, Micropropagation, Protocol, Wedelia chinensis.

Location: Department of Botany University of Chittagong Chittagong-4331, Bangladesh.

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: In vitro, Medicinal Plants

Objectives:

The present study was therefore undertaken with a view to developing an efficient and repeatable protocol for rapid and mass propagation of this medicinally important plant species with detailed evaluation of PGR combinations including those of both auxin and cytokinin group in the background of culture media.

Methodology:

MATERIALS AND METHODS A few Mahavringraj plants were collected from BCSIR Laboratories, Chittagong and were grown in earthen pots maintained in the departmental medicinal plant nursery. After one month of the plantation shoot apex, nodal segments and leaves of the healthy plants were collected and used as explant for in vitro experiments. The explants were washed thoroughly under running tap water for 30 min followed by treatment with savlon for 10 min. These were further washed thoroughly in running tap water for 10 min. The explants were finally surface sterilized with 0.1% (w/v) HgCl2 for 5 min followed by a dip in 70% ethanol for 30 s. These were then thoroughly rinsed five times with sterile distilled water. Before inoculation on to the culture media the explants were cut in to small pieces (1.0 cm apprx.) with the help of sterile surgical blade. MS basal medium supplemented with different concentrations and combinations of cytokinin (BAP and Kn) and auxin (IAA and NAA) were used for induction of organogenesis/embryogenesis. After 30 days of inoculation in culture room the response of the explants were thoroughly examined and the data on different aspects were recorded. For each treatment 15 explants were used and finally the data were subjected to statistical analysis for computation of standard error of mean (SE). For rooting, elongated shoot buds were cultured on rooting medium containing half strength MS medium fortified with different concentrations and combinations of IBA and IAA. In all the cases the media were solidified with the use of 0.8% (w/v) agar (Sigma) and pH was adjusted to 5.8 prior to autoclaving for 30 min at 121°C under a pressure of 1.1 kg/cm². All the cultures were incubated in a culture room at 25 ± 2°C under a regular cycle of 14 h light and 10 h dark. The complete seedlings thus produced by in vitro culture were finally planted in small earthen pots containing a mixture of soil and compost (2:1) through successive phases of acclimatization.

Source of Information: Journal of Medicinal Plants Research Vol. 5(11), pp. 2387-2392, 4 June, 2011 ISSN 1996-0875

Article Link (Online Journal): Available online at http://www.academicjournals.org/JMPR

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

After rooting the complete plantlets were transferred to outside earthen pots through successive phases of hardening. On average 80% of the plantlets transferred finally survived in the earthen pots which are now four months old and healthy (Figure 1E and F). It proves the possibility of using this protocol for rapid and mass propagation of W. chinensis. Although theprocess of direct organogenesis appears to be less effective in the use of genetic modification experiments but the emergence of direct gene transfer technology like electroporation, electrofusion, microinjection and microprojectile bombardment can be adopted for use of this process in the genetic improvement of this medicinal plant through in vitro technique.

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