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Development of an Efficient Regeneration System of Banana

M. M. Khatun
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

S. Islam
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

M. R. Kabir
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

D. Khanam
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

M. A. Y. Akhond
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

Abstract/ Executive Summary:

The present study was undertaken with a view to establish a reliable regeneration protocol of banana (Musa spp.). Flower buds, rhizome and apical dome sections were used as explant for callus induction. Explants were cultured on MS medium supplemented with different concentrations of 2, 4-D singly and in combination with different concentrations of IAA and NAA. Among the jive different treatments of 2, 4-D, T₅ (58%) showed better compared to other treatments in male flower bud explant. In rhizome and apical dome sections, all the treatment showed excessive swelling but no callus was observed upto seven weeks of culture initiation.

Keywords: Fruit, Banana, Flower buds, Rhizome, Apical dome

Location: Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur, Bangladesh

Start Date:

End Date:

Program Area: Farming System

Commodity: Banana

Objectives:

To develop an efficient regeneration protocol for banana which will be used for the development of banana transgenic with desirable traits.

Methodology:

Suckers of banana variety Amritsagar were used as source material. From the selected sucker a cube of tissue about 1-2 cm³ containing the apical meristem was excised. Firstly, these blocks of tissues were dipped in water, put 1-2 spoon of Ridomil gold MZ 68 WG, swirled with a spoon for 5 to 10 min. and washed in running tap water. Again these blocks were surface sterilized with jet and trix under running tap water for 15 min. followed by 3-4 times washing with distilled water. After that, these were taken inside the laminar air flow cabinet and put in sterile pot and dipped in Clorox solution (60 ml Clorox: 40ml distilled water) with 2 drops of Tween-20 for 20 minutes with shaking. Finally these were washed 3 to 4 times with autoclaved distilled water. The sterilized explants were excised with the help of sterilized scalpel and forceps. The dead tissue of sterilized block was cut off to leave a 5 x 8 rnm portion containing the apical dome covered with several leaf primordia together with 3 x 4 mm of rhizomatous bases. A thin layer of apical dome and rhizome sections were aseptically dissected. The prepared explants were placed in MS medium supplemented with 2, 4-D and four different concentrations of IAA and NAA. Another type of explants (male flower bud) were used. The fresh male flower buds (variety Sabri) were collected at the time of complete bunch emergence. Outer enveloping bracts were removed and surfaces were cleaned with absolute alcohol. Under laminar air flow cabinet, the remaining outer enveloping bracts were removed and the inner part (2-3 cm in length) containing male flower primordia were isolated. Individual male flower primordia were excidse and cultured on MS medium supplemented with different concentrations of 2, 4-D for initiation of callus. The p^H of all media was adjusted to 5.8 before autoclaving. The media were autoclaved at 121°C and 1.06 kg cm² pressure for 20 minutes. All cultures were incubated at 2.5±2°C in complete darkness and maintained on the same medium without subculture until callus formed.

Source of Information: Annual Research Report (2013-2014), Biotechnology Division, BARI, Gazipur, Bangladesh

Article Link (Online Journal):

Funding Source:

1.

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Male flower buds cultured on MS medium supplemented with five different concentrations of 2, 4-D showed enlargement of the floral primordia and the development of whitish callus after two and half months of inoculation. Results showed that bottom porting responded better. Among all the treatments, maximum callus was found in T₅ treatment (58%) followed by T₄ treatment (20%). Rest of the treatments showed only swelling of explant, no callus was found upto 75 days. Rhizome and apical dome section culture on different concentrations and combinations of 2, 4-D, IAA and NAA were shown. Result showed that in all the treatments explants exhibited swelling within four to five weeks. It was observed that upto seven weeks the explant showed excessive swelling but no callus was observed.

Knowledge Management: Report/Proceedings