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In Vitro Shoot Proliferation and Plant Regeneration of Physalis Minima L. - a Perennial Medicinal Herb

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.

Nadira Begum
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh.

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

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

Abstract/ Executive Summary:

The present study describes a protocol for high frequency plant regeneration of Physalis minima. Shoots were induced by culturing nodal segments and shoot tips from 15 day old seedlings. About 29 and 32 shoots were found to be induced from nodal segment and shoot tip explants, respectively, cultured on MS medium supplemented with 1.0 mg/l BAP. When shoots were subcultured on the fresh medium with same component as mentioned above, the shoots were elongated. Shoots rooted well when they were excised individually and implanted on half-strength MS medium with 0.3 mg/l NAA, where 98% shoots rooted within 12-15 days. In vitro grown plantlets with strong root system were successfully established in normal room temperature for seven days before transplanting in pots where they were reared for three weeks through successive acclimatization. The regenerated plants were successfully transferred to the soil with 90% survival rate.

Keywords: Physalis minima, Medicinal plant, Shoot proliferation, Micropropagation, Regeneration

Location: Biological Research Division, BCSIR Laboratories, Dhaka

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Non-commodity: Performance

Objectives:

The present communication deals with the development of a protocol for in vitro asexual multiplication in Physalis minima L.

Methodology:

The shoot tips and nodal explants of Physalis minima L. were collected from the medicinal plant garden of BCSIR Laboratories, Dhaka and used as explants for this experiment. The explants were washed thoroughly under running tap water, pre-soaked in liquid detergent (0.1% Tween-20) for about 30 min, washed three times again by running tap water 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 sterile distilled water in 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. Theexplants were placed vertically on the culture medium. For shoot induction and proliferation agar gelled MS (Murashige and Skoog 1962) basal media supplemented with BAP and coconut water (CW) at varying concentrations were prepared. Shoots induced from the in vitro cultures were further used as explants for adventitious shoot proliferation. For in vitro rooting, individual shoots (3-5 cm) were excised from the proliferated shoot cultures and implanted on half strength MS media with different concentrations and combinations of IBA, NAA and BAP. All media were supplemented with 30 g/l sucrose and 7 g/l agar (Difco). The pH of each medium was adjusted to 5.8 before autoclaving at 121^OC for 20 min and the medium was dispensed into 25x150 mm culture tubes or 250 ml conical flasks. The cultures were incubated for 16 h photoperiod at 24 ± 2^OC under a fluorescent light (3000 lux). Visual observation of culture was made every week. Data on shoot proliferation and root induction were recorded after three weeks of inoculation and used for calculation. For each treatment 15 explants were used for shoot proliferation and 15 shoots were used for rooting. All the treatments were repeated thrice. Data were analyzed statistically according to Mian and Mian (1984). 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.

Source of Information: Bangladesh J. Sci. Ind. Res. 44(4), 453-456, 2009

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

After four weeks the rooted shoots were transferred to pots. None of the plantlets survived when directly transferred from rooting medium to the pot under natural conditions. About 80 percent of the transplanted plants of Physalis minima survived if the plants in the rooting culture tubes were kept in normal room temperature for seven days before transplantation in pots and reared for three weeks. The plantlets were reared under semi-controlled temperature (30 ± 2^OC) and light (2000 lux) in a chamber with 80 percent humidity. During this period of acclimation shoots elongated, leaves expanded and turned deep green looking good and healthier

Knowledge Management: Journal