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In Vitro Regeneration for Mass Propagation in Commercial Scale of Medicinal Plant Vitex Nigundo L.

Md Abu Hena Mostofa Jamal
Department of Biotechnology and Genetic Engineering,Islamic University, Kushtia 7003, Bangladesh

A N M Rubaiyath-Bin Rahman
Department of Biotechnology and Genetic Engineering,Islamic University, Kushtia 7003, Bangladesh

Dipak Kumar Paul
Department of Applied Nutrition and Food Technology, Islamic University, Kushtia 7003, Bangladesh

Md Rezuanul Islam
Department of Biotechnology and Genetic Engineering,Islamic University, Kushtia 7003, Bangladesh

Abstract/ Executive Summary:

Context: It is necessary to focus on the importance of adopting micropropagation technique for mass propagation of the plantlets in commercial scale as well as conservation and distribution of germplasm. Objective: The present investigation has been designed with a view to establishing protocol of in vitro regeneration of medicinal plant species i,e., Vitex nigundo L (Verbenaceae). Materials and Methods: Shoot tips and nodal segments were used for multiple shoot induction. All explants were cultured on MS medium supplemented with various plant growth regulators. HgCl₂ was used as surface sterilizing agent. For in vitro rooting, individual shoots (3-4 cm) were cut from the proliferated shoot cultures and implanted on half and full strength of MS with different concentrations and combinations of NAA and IAA. The cultures were incubated for 16 h photoperiod at 25 ± 2ºC 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. Established plantlets were transplanted in earthen pots under natural conditions and the survival rate was recorded. Results: The most effective surface sterilization treatment has been found 0.1 % HgCl₂ for 7 minutes. Highest number of shoot was observed in MS medium supplemented with 3.0 mg/ BAP. It was rooted well in full MS containing 2.0 mg/l IAA. The survival rate was 85 % and propagated plantlets were successfully acclimatized in soil. Conclusion: It was observed that shoot tips are more responsive for micropropagation of Vitex nigundo L. Thus the fruitful utilization of rapid clonal propagation, germplasm conservation and distribution of Vitex nigundo, important medicinal plant of Bangladesh, is possible.

Keywords: Vitex nigundo, Medicinal plant, Shoot induction, Micropropagation, Regeneration.

Location: Department of Biotechnology and Genetic Engineering,Islamic University, Kushtia

Start Date:

End Date:

Program Area: Resource Development and Management

Non-commodity: Performance

Objectives:

The present investigation has been designed with a view to establishing protocol of in vitro regeneration of medicinal plant species i,e., Vitex nigundo L (Verbenaceae).

Methodology:

V. nigudo plants collected from the wild population of Jhenidah, Bangldesh were used as the explant sources for micropropagation. Different kinds of plant growth regulators i.e., BAP (6-benzyl amino purine), zeatin of cytokinin group and NAA (Napthalene acetic acid), IAA (Indole-3 Acetic Acid), 2,4 D (2,4-dichlorophenoxy acetic acid) of auxin group were used for this experiment. HgCl₂ was used as surface sterilizing agent. For plant growth MS medium containing nutrient basal salts were used which content macro, micro elements and vitamins. The explants were washed thoroughly under running tap water, presoaked in liquid detergent for about 30 min. They were then surface sterilized with 0.1% (w/v) mercuric chloride for 7 min, followed by five rinses with sterile distilled water inside a clean ench. The surface sterilized explants were sized to 3-4 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 (Murashige and Skoog 1962) basal media supplemented with BAP (6-benzyl amino purine), zeatin were prepared. The new shoots induced from the in vitro cultures were further used as explants for adventitious shoot regeneration. For in vitro rooting, individual shoots were cut (3-4 cm) from the proliferated shoot cultures and implanted on half and full strength of MS with different concentrations and combinations of NAA and IAA. All media were supplemented with 3% sucrose and gelled with 1% agar. Prepared media were dispensed into test tube (150×25 mm). The pH of the media was adjusted to 5.8 before autoclaving at 121°C for 20 min. The cultures were incubated for 16 h photoperiod at 25 ± 2°C 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. Established plantlets were transplanted in earthen pots under natural conditions and the survival rate was recorded.

Source of Information: J. bio-sci. 18: 140-145, 2010 ISSN 1023-8654

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The present work demonstrated the possibility of micropropagation of Vitex negundo through in vitro methods. Considering the importance of medicinal and other aromatic properties of V. negundo, micropropagation protocols can be effectively used in this species for various applications such as in vitro conservation, cryopreservation, large scale multiplication and genetic transformation. Thus the fruitful utilization of rapid clonal propagation, germplasm conservation and distribution of Vitex nigundo, important medicinal plant of Bangladesh, is possible.

Knowledge Management: Journal