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Regeneration of Medicinal Plant Paederia Foetida Through Direct Organogenesis Using Nodal Explants

S. H. Binto
Dept. of Crop Botany, Bangabandhu Sheikh Mujibur Rahman Agricultural University.

J. U. Ahmed
Dept. of Crop Botany, Bangabandhu Sheikh Mujibur Rahman Agricultural University.

T. K. Ghosh
Dept. of Crop Botany, Bangabandhu Sheikh Mujibur Rahman Agricultural University.

Abstract/ Executive Summary:

Chinese fever vine (Paederia foetida L.), a valuable medicinal plant has been greatly utilized in therapeutic purposes throughout the world. Since conventional propagation techniques of P. foetida are very slow, inefficient, and cannot cope with the increasing demand, in-vitro regeneration through tissue culture could be an alternative means of rapid propagation. Therefore, efforts were made to develop a suitable protocol through direct organogenesis of P. foetida. After surface sterilization, the nodal explants were cultured in Murashigue and Skoog (MS) medium and MS medium supplemented with different concentrations and combinations of plant growth regulators. MS medium supplemented with 6-benzyl amino purine; BAP (2.0 mg L^-1) produced the maximum number of shoots; 4.40 ± 0.98 and 5.40±1.12 after 15 and 30 days of culture respectively. The number of shoots gained by 15 days was found to be the highest; 1.20±0.80 at BAP (4.0 mg L^-1) followed by 1.00±0.55 at BAP (2.0 mg L^-1). Although the combination of BAP + Kinetin (2 mg L^-1 +2 mg L^-1) showed the highest shoot growth (3.40 ± 1.08 cm) by 15 days, sole application of BAP (2.0 mg L^-1) or Kn (0.5, 1.0, 2.0 and 3.0 mg L^-1) showed similar responses. BAP (2.0 mg L^-1) showed the best responses for developing the highest number of leaves; 18.60 ± 2.42 and 29.20 ± 2.73 respectively after 15 and 30 days of culture. Similarly, the development of the maximum number of leaves (10.60 ± 0.68) was reported by 15 days at BAP (2.0 mg L^-1). Rooting was significantly induced in indole-3-acetic acid (IAA) supplemented to 1/2 strength MS medium as compared to control (only ½ strength MS medium). The best performance of rooting was observed by 0.5 mg L^-1 IAA which produced average of 4.33 roots per shoot after 21 days of culture. The regenerated plants showed similar morphology to the mother plants. Thus, a suitable protocol for successful multiplication of P. foetida in vitro was established using nodal explants.

Keywords: Explant, Regeneration, Organogenesis.

Location: Department of Crop Botany, Bangabandhu Sheikh Mujibur Rahman Agricultural University.

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

To develop a suitable protocol for direct organogenesis of P. foetida in-vitro using nodal explants with different concentrations and combinations of growth regulators.

Methodology:

sterilization and culture of plant materials About 1.0 cm of healthy and juvenile nodal explants were collected from donor P. foetida plants grown at the Department of Crop Botany, BSMRAU. Then, those explants were thoroughly washed with running tap water 5 to 6 times, followed by washing with Tween-20 solution for 5 minutes and distilled water for at least 3 times. After that, surface sterilization was done by submitting the explants in 70% ethanol for 5 mins followed by 1% sodium hypochlorite (NaOCl) for 2 mins and 0.1% mercuric chloride (HgCl₂) for 2 mins respectively. Then the explants were washed repeatedly with sterile distilled water and allowed for culture in freshly prepared MS (Murashige and Skoog, 1962) medium or medium fortified with different growth regulators for showing shooting responses. For root initiation, ½ strength MS medium and ½ strength MS medium supplemented with different concentrations of indole-3-acetic acid (IAA) were used. A 16 h photoperiod at 25±2 ºC under the illumination of 35-50 μmolm^-2s^-1 was used to incubate the cultures as described by Thirupathi et al. (2013). treatment combinations MS medium or MS medium supplemented with different concentrations and combinations of plant growth regulators viz.6-benzyl amino purine (BAP) and kinetin (Kn) either sole or in combinations were used to find out the suitable growth regulator or combination of growth regulators for direct organogenesis of P. foetida using nodal explants. For the observation of shooting responses, MS medium without growth regulators was treated as a control (0.0). MS medium supplemented with BAP (0.5, 1.0, 2.0, 3.0 and 4 mgL^-1), Kinetin; Kn (0.5, 1.0, 2.0, 3.0 and 4 mgL^-1) and combined application of BAP and Kn (0.5+0.5, 1.0+1.0, 1.5+1.5, 2.0+2.0 and 4.0+3.0 mgL^-1) were used as different treatment combinations. For rooting, ½ strength MS medium was used as control (0.0) and ½ strength MS medium supplemented with different concentrations of indole-3-acetic acid, IAA (0.2, 0.5, and 1.0 mgL^-1 ) was used as treatment. data collection and analysis regular observation was made in the growth and shooting pattern of the explants in in-Vitro and data regarding the number of shoots, shoot length, and the number of leaves were collected after 15 and 30 days after culture. The number of root initiation from the explant was recorded after 14 days and 21 days of culture. The experiment was set by following Completely Randomized Design (CRD) with 5 replications. The data were analyzed by following the Statistics 10 package computer programme. Mean separation using LSD values was calculated by the same programme. Student t-test was also used for showing the statistical differences.

Source of Information: Ann. Bangladesh Agric. (2020) 24(1) : 88-98

Article Link (Online Journal): DOI: https://doi.org/10.3329/aba.v24i1.51938

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Based on the findings of the present efforts, it can be concluded that MS medium supplemented with BAP (4.0 mg L^-1) was found to be very effective for the regeneration of new shoots. Combined application of BAP + Kn (2 mg L^-1 +2 mg L^-1) with MS medium showed the best responses for enhancing shoot length. MS medium supplemented with BAP (2.0 mg L^-1) showed the best efficacy for the development of new leaf. MS medium (1/2 strength) supplemented with IAA (0.5 mg L^-1) showed the best performance for initiation of roots. Although the protocol which we claimed here is likely to be useful for the large-scale multiplication of medicinal plant P. foetida, further efforts by engaging other plant parts and growth regulators should be needed to find out the best in-vitro regeneration strategy of P. foetida.

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