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In Vitro Shoot Multiplication of the Biodiesel Plant Jatropha Curcas L. Through Direct Organogenesis

M. A. K. M. S Hassan
Department of Botany, Savar College, Savar, Dhaka-1340,

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

L. S. Bari
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

M. A. A. Jahan
Biological Research Division, BCSIR Laboratories, Dhaka-1205, Bangladesh

Abstract/ Executive Summary:

An efficient protocol was established for in vitro shoot multiplication of the biodiesel plant, Jatropha curcas L. (Euphorbiaceae) through direct organogenesis using shoot tip and nodal explants. Best shoot induction was observed on MS basal medium supplemented with 1.5 mg/l BAP + 0.5 mg/l NAA, in which 86.2% of nodal explants responded to produce maximum number (7.2 ± 0.68) of shoots per culture. In vitro raised shoots rooted on half strength MS medium with 1.0 mg/l IAA. The survival rate of regenerated plantlets was 85%.

Keywords: Jatropha curcas; Biodiesel plant; Multiplication; Organogenesis; Acclimatization

Location: Medicinal Plants Garden of Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Non-commodity: Performance

Objectives:

To develop a protocol for in vitro shoot multiplication of the biodiesel plant, Jatropha curcas L. through direct organogenesis using shoot tip and nodal explants.

Methodology:

Jatropha curcas L. at Medicinal Plants Garden of Bangladesh Council of Scientific and Industrial Research (BCSIR), Dhaka, was used as a source of explants. Shoot tip and nodal explants with a single axillary bud were used for this purpose. The explants were washed thoroughly under running tap water, pre-soaked in Tricks 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 times rinse with sterile distilled water under laminar air flow cabinet. The surface-sterilized explants were cut into 1-1.5 cm 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. The new shoots induced from the in vitro cultures were further used as an explants for adventitious shoot regeneration. MS (Murashige and Skoog, 1962) basal medium was used for shoot proliferation and adventitious shoot regeneration and half strength MS was used for in vitro root induction. All media were supplemented with 30 g/l sucrose, 7 g/l agar (Difco) and dispensed into 15×150 mm culture tubes and 250 ml conical flasks. The pH of the media was adjusted to 5.8 before autoclaving at 1.9 kg/cm2 pressure at 121°C for 20 min. The cultures were incubated for a 16 h photoperiod at 24 ± 2°C under 1200 lux/m2 fluorescent light. Shoot proliferation from shoot tip and nodal explants was obtained in two separate sets of experiments. In the first experiment 0.1-2.0 mg/L BAP were incorporated into MS media to select the best cytokinin for the response of shoot induction. In the second set, combination of BAP (0.5-2.0 mg/L) with NAA (0.1-0.5 mg/L) and BAP (0.5-2.0 mg/L) with IAA (0.1-0.5 mg/L) were assessed for shoot multiplication. Number of new proliferated shoot of in each culture was recorded after every week of inoculation. For in vitro rooting, individual shoots (3-5 cm) were excised from the proliferated shoots and implanted onto half strength MS with different concentrations and combinations of NAA, IBA and IAA. The rooted plants were washed to remove agar gel adhered to the roots and transplanted to plastic pots with soil and compost (1: 1) for hardening. The plantlets were kept in a polychamber at 80% relative humidity, 32 ± 2°C temperatures for a 12 h photoperiod under 1500 lux/m2 sun light for acclimation. Established plants were transplanted in earthen pots under natural conditions and the survival rate was recorded. All experiments were repeated three times. Each treatment had 15 replicates. The morphologenetic response of explants for microshoot induction was evaluated in eight weeks of culture. For microshoots proliferation and plantlet formation, results were evaluated in eight weeks of culture. Morphogenetic response was expressed as percentage of explants with microshoots in relation to the number of surviving explants. For acclimatization 50 or 25 plantlets were taken for each treatment. Data were statistically analyzed and in some parameters means were compared using DMRT (Duncan, 1955).

Source of Information: Bangladesh J. Sci. Ind. Res. 49(1), 41-46, 2014

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

About 85 per cent of the transplanted plants of Jatropha curcas 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 ± 2ºC) and light (1500 lux) in a chamber with 80 per cent humidity. During this period of acclimation shoots elongated, leaves expanded and turned to deep green and healthier. Following the transfer of plants to an open place and gradually acclimatized to outdoor conditions, where 85 per cent plants were survived. The technique described here appears to be readily adaptable for large scale plant regeneration and plantation of Jatropha curcas.

Knowledge Management: Journal