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Plant Regeneration from Azadirachta Indica A. Juss Nucellar Callus

R. Islam
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

M. A. Hoque
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

O. I. Joarder
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

Abstract/ Executive Summary:

Callus cultures of nucellar tissues excised from fertilized ovules of immature fruits of Azadirachta indica were obtained in MS medium containing 2.5 - 5.0 μM benzyladenine (BA) and 5.0 - 20.0 μM naphthalene acetic acid (NAA). Compact nodular calli, upon transfer to media containing 5.0 - 20.0 μM BA and 1.0 - 5.0 μM NAA, produced adventitious shoots. Isolated shoots grown in the medium with 0.5 μM gibberellic acid and 0.1 μM BA elongated into healthy shoots. Rooting of in vitro regenerated shoots was obtained in half-strength of MS fortified with 5.0 μM indole-3-butyric acid (IBA). The regenerants were transferred successfully to soil after acclimatization with 70% survival.

Keywords: Nucellus culture, Adventitious shoots, Neem

Location: Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Indian lilac (Neem)

Objectives:

To describes a method of multiplication of A. indica from callus cultures derived from nucellar tissues.

Methodology:

Immature fruits (30 - 40 d after pollination) were collected from a 20- year-old A. indica tree. Seeds were removed and washed thoroughly under running tap water and surface disinfected by immersing in 0.1% HgCl₂ for 12 min and washing three times with autoclaved distilled water. Under aseptic conditions nucellar tissues were dissected out excluding the zygotic embryos. These nucellar explants, were induced for callus initiation on MS medium supplemented with 5.0 - 20.0 μM NAA for 2.5 - 5.0 μM BA. After five weeks, calli derived from nucellar tissues were transferred to the same basal medium supplemented with 5.0 - 20.0 μM BA and 1.0 - 5.0 μM NAA for shoot proliferation. The shoots elongated in the medium with 5.0 μM gibberellic acid (GA₃) and 0.5 μM BA. The elongated shoots were rooted in half- strength MS media with 5.0 μM IBA. The medium pH was adjusted to 5.7 prior to sterilization by autoclaving and solidified with 0.7% Difco Bacto-agar. The cultures were maintained at 26 ± 1°C at a relative humidity of 55 - 60% under a 16 h photoperiod (50 - 70 μMol m^-2s^-1) provided by warm white fluorescent tubes. All experiments were repeated twice and mean confidence intervals were obtained at p = 0.05 using t-test.

Source of Information: Plant Tissue Culture, Vol: 8, (1), pp: 69-75, ISSN 1018-8029, (June) 1998

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The present study demonstrates the formation of complete plantlets from the callus cultures derived from nucellar tissues of A. indica. This is the first report of plant regenertion from nucellar tissues of this species. In vitro plant regeneration from nucellar explants has been described. Nucellar derived plants are free of virus and other disease causing organisms due to absence of vascular connections between the surrounding maternal tissue and the nucellus. The nucellus culture technique has been used to recover virus free citrus cultivars in Spain. Efficient recovery of A. indica plants through the process described here will facilitate rapid propagation of horticulturally useful disease-free plants and is potentially useful for international exchange of germplasm.

Knowledge Management: Journal