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In Vitro Clonal Propagation of the Neem Tree (azadirachta Indica A. Juss.)

M. Shahin-uz-zaman
MS Student
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. Ashrafuzzaman
Professor
Department of Crop Botany, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. Shahidul Haque
Professor
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Lutfun Nahar Luna
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

A study was conducted at Biotechnology Laboratory of Bangladesh Agricultural University, Mymensingh with root and shoot tip explants of neem to develop an efficient protocol of regeneration. Shoot tips and root tips from 10 - 20 days old seedlings of neem were cultured on (MS) medium supplemented with different concentrations and combinations of BAP (0.0, 0.1, 0.2, 0.3, 0.4, 0.5 and 0.6 mg l^-1) and NAA (0.0 and 0.05 mg l^-1). Shoot buds were initiated from the both explants. The highest percentage of regeneration was found from shoot tip but the highest average number of shoots/explants was found from root tip explants. The regenerated shoots were further sub-cultured and later could be rooted on a medium supplemented with different concentrations and combinations of IBA and IAA and complete plants could be obtained. The rooted plants were transplanted to pots for hardening.

Keywords: Neem, Root tip, Shoot tip, Explants, Indole-butyric acid, Indole-3-acetic acid

Location: Biotechnology Laboratory of Bangladesh Agricultural University, Mymensingh

Start Date: 00-00-2005

End Date: 00-00-2005

Program Area: Variety and Species

Commodity: Indian lilac (Neem)

Objectives:

To develop an efficient protocol of regeneration.

Methodology:

A study was conducted at Biotechnology Laboratory of Bangladesh Agricultural University, Mymensingh with root and shoot tip explants of neem to develop an efficient protocol of regeneration. Immature fruits of A. indica were collected from 30 years old tree grown in Bangladesh Agricultural University Campus. Freshly collected immature fruits were washed thoroughly with distilled water and also surface sterilized by 0.1% HgCl₂ for 5 min followed by washing with sterile water for five times. Excised cotyledons were then aseptically incubated in flasks containing agar gelled MS medium. After 30 to 40 days root tip and shoot tip of seedlings were dissected out and cultured on MS medium supplemented with various concentrations and combinations of 6-benzyl aminopurine (BAP) and naphthalene acetic acid (NAA) for shoot initiation. The culture vials were transferred to growth room and were allowed to grow in controlled environment. The temperature of the growth room was maintained with in 25±2°C by an air conditioner. A 16 h light period was maintained with light intensity of 2000 lux of the growth and development of culture. The primary shoots from shoot tip and root tip explants cultured on MS medium incorporated with 0.1 - 0.6 mg l^-1 BAP and 0.05 mg l^-1 NAA and thereafter regenerated microshoots were excised several times and cultured on different media. For successful rooting, 3 - 4 cm usable shoots excised from multiplication culture were implanted to rooting medium consisting of 1/2 MS medium supplemented with IBA and/or IAA. One-month-old adequately rooted shoots when reached a height of 4 - 5 cm were brought out from culture room and kept in room agar was washed out gently from roots. These plantlets were transplanted in small pots contained compost, soil and sand (1:2:1) and were covered with transparent sheets. They were nurtured under room temperature in presence of sufficient light. An average days nursing under room condition enabled them to adapt in outdoor conditions. In that conditions the plantlets were watered every alternate day.

Source of Information: African Journal of Biotechnology Vol. 7 (4), pp. 386-391, 2008

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Root tip explants are best for multiplication of neem. Among hormone concentrations, T₃is best for shoot induction and multiplication and IBA at 0.05 mg l^-1 + IAA at 0.05 mg l^-1 are best for rooting.

Knowledge Management: Journal