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Micropropagation of Date Palm (phoenix Dactylifera L.) (2019--2020)

S.C. Halder
Biotechnology Division, BARI, Joydebpur, Gazipur

M.M. Khatun
Biotechnology Division, BARI, Joydebpur, Gazipur

M.K.Hasan
Biotechnology Division, BARI, Joydebpur, Gazipur

M.A.Y. Akhond
Biotechnology Division, BARI, Joydebpur, Gazipur

Abstract/ Executive Summary:

The aim of this study was to develop an efficient micropropagation protocol of date palm. Offshoots and inflorescence were used as explants sources. Different concentrations of IAA, NAA and 2,4-D were used for initial culture establishment. In offshoot explant, organogenesis responses were noticed after 2 months of culture in the same fresh medium. In inflorescence explant, some floret showed pro-embryogenic masses and some swelled in size. But still, now, no embryos or shoots were found.

Keywords: Laminar airflow, Arabian date palm, Offshoot explants, Inflorescence explant

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Date

Objectives:

Micropropagation of date palm has become a preferred alternative over offshoot propagation for large-scale production to satisfy the market demand to undertake micropropagation to overcome the obstacles.

Methodology:

Offshoots of Arabian date palm were collected from the date palm garden of BRRI, Gazipur and used as an explant source. Arabian Jumbly and Barhee varieties were used. Healthy offshoots (Fig. 1a) were separated from field-grown trees using sharp tools. Older leaves were trimmed off. Leaf fibers were cleaned from the base and the leaf sheaths were removed one by one from the outer ring towards the centre. The final size of the offshoot head was about 2 to 3 cm in width and 4 to 6 cm in length. The prepared explants (Fig. 1b) were surface sterilized with Ridomil Gold 68 WG for 30 minutes to remove the fungal contamination. Then shoot tips were washed with trix and detergent for 25 minutes. After that shoot tips were sterilized with Clorox (100%) by adding 4-5 drops of Tween-20 for 20 minutes followed by 4-5 times washing in sterilized distilled water. Sterilized explants were kept in a cold sterilized solution of ascorbic acid and citric acid (150 mg/l) for 10 minutes to avoid browning. Sterilization and final dissection were carried out under a laminar airflow cabinet in aseptic condition. The final prepared explants were cut into small pieces and placed in MS medium supplemented with different concentrations of IAA, NAA and 2, 4-D. All media had 3% sugar and 0.8% agar. The pH of all media was adjusted to 5.6 before autoclaving. The media were autoclaved at 121⁰C and 1.06 kg cm² pressure for 20 minutes. All cultures were incubated in complete darkness till callus formation. The inflorescence (spathe) was excised from the mother plant, placed in a clean plastic cover and brought carefully from the open field to the laboratory. For surface sterilization, the intact spathe has been dipped in fungicide solution Ridumil Gold 68 WG for 30 seconds without any shaking. The spathe was washed under running tap water for 1 min. The water streams were directed at the basal part of the spathe to remove dirty material. After that, the spathe was sterilized with 60% sodium hypochlorite (NaOcl) solution by adding 4 drops of Tween-20 for 5 min. The spathe was finally washed with sterilized distilled water 3 times for 1 min without shaking. Then, the spathe was cut longitudinally from the middle like a T from only one side. The cut was made in the central swollen portion of the spathe due to its softness and the spikelets were taken out one by one with the help of forceps. The spikelets were cut into different sizes from their base. The cut explants are as follows: i) spikelets are 2.3 cm with 3 florets ii) spikelets are 4.4 cm with 4 florets iii) spikelets are 4.6 cm with 5 florets and iv) spikelets are 5.6 cm with 6 florets (Fig. 1e). Then, the explants were placed horizontally on the sloping surface of the nutrient medium. All cultures were incubated in a controlled growth chamber at 25 ± 2^oC in the dark. Incubated explants were subcultured every 3-4 weeks on the same culture medium. Well, responding explants were transferred to another medium.

Source of Information: Annual Research Report 2019-2020, Biotechnology Division, BARI, Joydebpur, Gazipur

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Offshoot explants of Arabian Jumbly were cut into small pieces and cultured on MS medium supplemented with different concentrations of IAA, NAA and 2,4-D for initial culture establishment. The explants were swelled and enlarged in sizes after one subcultured. Organogenesis responses were noticed after 2 months of culture in the same fresh medium. Only, root formation was observed. Some explant produced both roots and callus. They were then transferred to another medium. After 4 months, no responses were observed. In inflorescence explant, different cut spikelet sized with 3-6 florets were cultured on MS medium supplemented with different concentrations of IAA, NAA and 2,4-D. Subculture was done every 3-4 weeks. After one subculture, some floret showed blackened & died, some produced pro-embryogenic masses and some swelled, enlarged in size and produced callus. They were again subcultured in the same fresh medium. After 2-3 subculture, they were transferred to another medium which contains 2, 4-D and 2ip in basal MS medium. But still, now, no embryos or shoots were found.

Knowledge Management: Report/Proceedings