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In Vitro Regeneration of Vitex Negundo L., a Woody Valuable Medicinal Plant Through High Frequency Axillary Shoot Proliferation

Farhana Afroz
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

A.K. M. Sayeed Hassan
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

Laila Shamroze Bari
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

Rebeka Sultana
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

John Liton Munshi
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

Miskat Ara Akhter Jahan
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

Rahima Khatun
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

Abstract/ Executive Summary:

An efficient protocol was established for rapid and large scale propagation of woody aromatic medicinal plant Vitex negundo L. by in vitro shoot multiplication from shoot tips and nodal segments of mature plant. Of the four different growth regulators BA, Kn, GA₃ , NAA and coconut water, MS fortified with BA 1.0 mg/l was found to be the most effective for inducing multiple shoots from nodal explants. The percentage (96%) of shoot multiplication per node (21.83) was highest up to second subculture pas-sages, after which there was a gradual decline in shoot development. Best rooting was induced (93%) in excised shoots on half strength MS medium supplemented with an optimal combination of NAA (0.3 mg/l). Soil, compost and sand (1:1:1) mixture was the most suitable planting substrate for hardening. The survival rate was 80% and the regenerated plants were successfully transferred to the soil.

Keywords: Vitex negundo, Medicinal plant, Shoot proliferation, Micropropagation, Regeneration

Location: BCSIR Laboratories, Dhaka-1205

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Medicinal Plants

Objectives:

To development of a protocol for in vitro asexual multiplication in Vitex negundo.

Methodology:

Shoot tips and nodal explants of Vitex negundo L. collected from the garden of medicinal plants in the premises of BCSIR Laboratories, Dhaka-1205, were used for this experiment. The explants were washed thoroughly under running tap water, pre-soaked in liquid detergent for about 30 min, wiped with cotton and dipped in 70% (v/v) ethanol for 1 min. They were then surface sterilized with 0.1% (w/v) mercuric chloride for 5 min, followed by five rinses with ster-ile distilled water inside a laminar air flow cabinet. The surface sterilized explants were sized to 1.0-1.5 cm in length containing a single node with an axillary bud or a shoot tip with an apical bud. The explants were placed vertically on the culture medium. For shoot induction agar-gelled MS basal media supplemented with BA, Kn, GA₃ , coconut water (CW) at varying concentrations were prepared. The new shoots induced from the in vitro cultures were further used as an explants for adventitious shoot regeneration. For in vitro rooting, individual shoots (3-5 cm) were cut from the proliferated shoot cultures and implanted on half strength MS with different concentrations and combinations of NAA and IBA. All media were supplement-ed with 30 g/l sucrose and gelled with 7 g/l agar (Difco). Prepared media were dispensed into 150 x15 mm culture tubes or 250 ml conical flasks according to experiment. The pH of the media was adjusted to 5.8 before autoclaving at 121^oC for 20 min. The cultures were incubated for 16 h photoperiod at 24 ± 2^oC under a fluorescent light. Visual observation of culture was made every week. Data on shoot induction and proliferation and root induction were recorded after three weeks of inoculation and used for calculation. For each treatment 15 explants were used and all the treatments were repeated thrice. The healthy plantlets were taken out from the culture tubes, washed to make free from agar gel with running tap water and transplanted to plastic pots containing soil, sand and compost (1:1:1) for hardening. The plantlets were kept in a poly chamber at 80% relative humidity, 32 ± 2^oC under 12 h photoperiod for acclimation. Established plantlets were transplanted in earthen pots under natural conditions and the survival rate was recorded.

Source of Information: Bangladesh J. Sci. Ind. Res.43(3), 345-352, 2008

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The explants were found to be swollen and they produced two to five shoots within three-four weeks after inoculation. The highest percentage of shooting response of the inoculated explants was 96% with 21.83 shoots per culture when the explants were cultured in MS medium with 1.0 mg/l BA, which was followed by the media supplemented with 1.0 mg/l BA + 5% CW, 1.5 mg/l BA, 2.0 mg/l BA, 0.4 mg/l BA + 0.1 mg/l NAA, 0.1 mg/l BA and 0.2 mg/l Kn + GA₃. Both the explants responded in the same medium but the highest number of micro shoots were found to be induced in nodal explants.

Knowledge Management: Journal