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In Vitro Shoot Proliferation and Development of Micropropagation Protocol from Leaf Disc of Gladiolus

M. Shaheenuzzaman
Agronomy Division, BARI, Gazipur,

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

M. M. Karim
Agronomy Division, BARI, Gazipur,

Z. U. Noor
Farm Division, BARI, Gazipur, Email: kbdshaheen78@yahoo.com

Abstract/ Executive Summary:

The experiment was carried out during the period from July 2008 to March, 2009 at the Biotechnology Laboratory, Department of Biotechnology, Bangladesh Agricultural University, Mymensingh. The present investigation was to be better source for shoot multiplication. The calli derived from leaf discs were cultured on shoot induction media containing different combinations and concentrations of BAP (1.0, 1.5, 3.0 and 6.0 mg/L) and Kn (0.5, 1.0, 1.5 and 2.0 mg/L). The highest percentage of shoot regeneration (91.66%) and the highest shoot length was 4.24 (cm) with a minimum number of days (13.33) was observed in the MS medium supplemented with 3.0 mg/L BAP and 0.5 mg/L Kn. Thus shoot multiplication in successive subculture was possible. The present investigation was carried out to study the in vitro shoot multiplication in gladiolus by using leaf disc as explants.

Keywords: Shoot proliferation, Micropropagation, BAP: 6-benzylamino purine, Kn: Kinetin, Leaf disc

Location: BAU, Mymensingh

Start Date: 00-07-2008

End Date: 00-03-2009

Program Area: Variety and Species

Commodity: Gladiolus

Objectives:

1.To determine a suitable hormone combination shoot regeneration and plantlet establishment

2 To develop protocol for in vitro plantlet regeneration of Gladiolus

Methodology:

The experiment was carried out during the period from July 2008 to March, 2009 at the Biotechnology Laboratory, Department of Biotechnology, Bangladesh Agricultural University, Mymensingh. The planting materials (bulbs) of Gladiolus from four varieties ware collected from Germplasm Center, Bangladesh Agricultural Research Institute. The Bulbs were used to germinate plants in vitro condition and after three weeks, the leaves were used as explants. MS (Murashige and Skoog, 1962) medium supplemented with different phytohormones as treatments was used in culture medium for shoot induction, shoot multiplication and maintenance and regeneration of roots from multiplied shoot. The hormones wereadded separately to MS media. For the preparation of media, stock solutions were prepared at the beginning and stored at 4±1º C. The media were prepared from the stock solutions. The leaf derived calli were cultured on MS medium supplemented with combinations of various concentrations of BAP (1.0, 1.5, 3.0 and 6.0 mg/l) and Kn (0.5, 1.0, 1.5 and 2.0 mg/L) for shoot initiation. For the purpose of hardening and acclimatization to the field conditions, 10 plants (8-10 weeks old) were taken out of the culture vessels. The roots of the plant lets were thoroughly washed with tap water in order to remove even the traces of the nutrient medium. The plant lets were then transferred to the pots containing presterilized sand. The pots were kept in the culture room for two weeks; adequate water was given to the plant lets at an interval of two-to three days. After two weeks, the plants were shifted to the soil in pots and kept in the growth room. After further hardening of two weeks, the plants were shifted to the normal growth conditions of the field. In this study, an efficient and reproducible protocol for the plantlet regeneration from leaf discs explants in Gladiolus was developed. This protocol has the potential for the in vitro regeneration of Gladiolus.

Source of Information: J. Bangladesh Agril. Univ. 9(1): 21–26, 2011 ISSN 1810-3030

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The highest percentage of shoot regeneration (91.66%) and the highest shoot length was 4.24 (cm) with a minimum number of days (13.33) was observed in the MS medium supplemented with 3.0 mg/L BAP and 0.5 mg/L Kn. Thus shoot multiplication in successive subculture was possible. The present investigation was carried out to study the in vitro shoot multiplication in gladiolus by using leaf disc as explants.

Knowledge Management: Journal