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In Vitro Morphogenesis of Arabian Date Palm (phoenix Dactylifera L.)

Md. Abul Kalam Azad
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia

Hasnatul Arefin
Biotechnology Division, Bangladesh Sugarcane Research Institute, Ishurdi, Pabna

Md. Amzad Hossain
Biotechnology Division, Bangladesh Sugarcane Research Institute, Ishurdi, Pabna

Abstract/ Executive Summary:

After inoculation of young leaves of date palm offshoot required about six months to complete the morphogenesis process. Fourteen weeks were required for embryogenic callus formation under continuous dark condition and nine weeks for shoot initiation (under 16/8 h light/dark). The highest number of explants (80%) produced callus in modified MS containing 5 mg/l 2,4‐D + 2 mg/l 2ip. Sixty per cent of explants produced callus in the modified medium containing 5 mg/l 2,4‐D + 5 mg/l NAA. While only 50 per cent of the explants formed callus in the same medium when supplemented with only 5 mg/l 2,4‐D. The induced calli were transferred to modified MS for shoot proliferation. A combination of two cytokines showed better performance than single ones in shoot induction. The highest percentage (70) of shoot developed in modified MS containing 2 mg/l BAP + 1 mg/l Kn. Forty per cent shoot induction was found in the same medium supplemented with 2 mg/l of BAP. Thirty per cent shoot formed in MS containing 1 mg/l of Kn. The shoots were subcultured at three four week intervals throughout culture duration.

Keywords: Arabian date palm, morphogenesis, Juvenile leaf, Morphogenesis

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Date palm

Objectives:

The objective of this study was to investigate the role of different concentrations and combinations of 2,4‐D, NAA, 2ip, BAP and Kn to induce somatic embryogenesis and shoot initiation from juvenile leaf explants of date palm.

Methodology:

An efficient morphogenesis protocol has been developed through in vitro culture technique from juvenile leaves of offshoot of date palm were used in various combinations and concentrations of PGR in modified MS. This is the first report of in vitro date palm morphogenesis in Bangladesh. Offshoots were obtained from adult Arabian date palm cv. Ajwah growing in Bangladesh Sugarcane Research Institute (BSRI) date palm garden. About 6 ‐ 8 cm long leaf base was cut from the unexpanded leaf. These collected parts were first washed thoroughly in running tap water for 10 ‐ 15 min, then they were cleaned with 5% Savlon and 70% ethyl‐alcohol and rinsed with sterile double distilled water. Thereafter then they were cleaned with liquid detergent Tween 20 (1% v/v) for 5 ‐ 10 min and rinsed with sterile double distilled water. Finally they were surface sterilized with 0.1% HgCl₂ (w/v) solution for 20 min and again washed well in sterile distilled water three ‐ four times to remove all traces of HgCl₂(Badawy et al. 2005). For all the above studies, modified MS contained 3% (w/v) sucrose, 0.5 g/l activated charcoal (AC), 10% coconut milk, NaH₂PO₄(170 mg/l), citrate (100 mg/l), biotin (2 mg/l) and were solidified with 0.7% agar. The pH of all media was adjusted to 5.7 prior to autoclaved at 121ºC at 15 psi for 20 min. Experiments were repeated at least three times and at least ten cultures were employed per treatment (Jain and Gupta 2005).

Source of Information: Plant Tissue Cult. & Biotech. 23(2): 211-219, 2013 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The embryogenic calli derived from young leaves of offshoot were transferred to shoot induction medium (modified MS) supplemented with different concentrations of BAP and Kn alone and or in combination. The embryogenic calli were kept in 16 hrs light conditions. The highest percentage of shoots (70) were regenerated from embryogenic calli when cultured in modified MS containing 2 mg/l BAP + 1 mg/l Kn. Forty per cent shoots were formed in modified MS containing only BAP @ 2 mg/l while only 30 per cent shoots were formed in the same medium containing 1 mg/l Kn. It was revealed that shoot induction percentage increased up to 2 mg/l BAP and or up to 1 mg/l Kn, and declined the percentage of shoot formation beyond the above concentration. Therefore, it could be mentioned that the combined effect of different cytokinins showed better performance than single one for shoot induction from embryogenic calli. Two cytokinin tested BAP was more active than Kn during multiple shoot formation. The superiority of BAP than Kn for multiple shoot formation was also demonstrated in other plants like Jetropha integerrima and Bombaxceiba. Shoot multiplication rate decreased with increasing BAP concentrations up to 3 mg/l. Similar effect was also found in many other plant tissue cultures. Present study elucidated an efficient protocol for plant morphogenesis via embryogenesis of date palm and this will be helpful in conducting micropropagation.

Knowledge Management: Journal