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Efficient Plant Regeneration from Cotyledon and Midrib Derived Callus in Eggplant (solanum Melongena L.)

M Rahman
Department of Genetic Engineering and Biotechnology, University of Rajshahi, Rajshahi-6205, Bangladesh.

M Asaduzzaman
Biotechnoloqy Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh.

N Nahar
Biotechnoloqy Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh.

M A Bari
Biotechnoloqy Laboratory, Institute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh.

Abstract/ Executive Summary:

Somatic embryos were obtained from cotyledon and midrib explants of Solanum melongena L., cultivar Loda. For callus induction, medium was supplemented with different concentrations of auxin singly or in combination with BAP. The best callusing 83 - 85% was obtained from both of the explants cultured on MS medium containing 2.0 mgl^-1 NAA + 0.05 mgl^-1 BAP. Somatic embryogenesis and shoot regeneration was achieved after transferring the calli to MS medium supplemented with BAP, GA₃, NAA and Zeatin. Cotyledon derived calli showed better performance (87%) for regeneration than that of midrib (82%) when sub cultured on MS medium having 2.0 mgl^-1 Zeatin + 1.0 mql^-1 BAP. For root induction, MS + 3.0 mql^-1IBA was proved to be better treatment for average number (14-15) and mean length (12 cm) of roots than those of other treatments.

Keywords: Eggplant, Cotyledon, Midrib, Callus induction, Somatic embryo

Location: Rajshahi University, Rajshahi

Start Date: 01-01-2000

End Date: 30-06-2001

Program Area: Variety and Species

Commodity: Brinjal

Objectives:

To establish a protocol for efficient plant regeneration from callus culture in eggplant, cultivar Loda using cotyledon and midrib as explants.

Methodology:

Loda cultivar of eggplant (Solanum melongena L.), commonly cultivated in the northern region of Bangladesh, was selected for the study. Seeds were aseptically grown in culture vessels. For initial establishment of callus culture, 3 weeks old in vitro grown seedlings were used as the source of explants. Cotyledon and midrib isolated from the in vitro grown seedlings were cultured for callus induction and subsequent somatic embryogenesis. Cotyledon and midrib explants were taken from the in vitro grown plants of 3 weeks old. Both explants were excised with the aid of scalpel blade and collected in a petridish. Then the explants were laid on the sterile tile using sterile forceps and were cut into segments of 2 - 3 mm with the help of a scalpel blade. The explants were then placed on the medium having MS nutrients supplemented with various concentrations and combinations of 2, 4 - D, NAA, IAA and BAP with 3% sucrose and solidified with 0.7% agar. The pH was adjusted to 5.8 before autoclaving at 121°C for 20 min. The explants were cultured in petridishes (9 cm) and maintained in the culture room at 25 ± 2°C under a 16h photoperiod (cool- white fluorescent tube supplying). Each petridish contained 18- 20 pieces of explants for both cotyledon and midrib. After 4 weeks, the induced calli were sub- cultured on fresh medium with new hormonal supplements for further proliferation and somatic embryogenesis. Plants were regenerated by transferring the suitable embryogenic calli in MS semisolid medium supplemented with 2, 4- D, NAA, GA₃, BAP and Zeatin at various concentrations and combinations. Within 3- 4 weeks of inoculation, masses of 15- 20 shoots were developed from each callus, which were sub divided and cultured separately for further proliferation and root induction. The cultures were inoculated at 25 ± 2°C under 16/ 8 h light/ dark condition. Proliferated shoots should have proper root development for successful establishments of plant lets in natural condition. Shoots of 2- 3 cm were placed in MS medium supplemented with different concentrations and combinations of IBA and GA₃.MS₀ and 1/2 strength of MS were also treated for root induction.

Source of Information: JOURNAL OF BIO-SCIENCE, THE INSTITUTE OF BIOLOGICAL SCIENCES RAJSHAHI UNIVERSITY, Vol. 14, 2006, pp. 31-38, ISSN 1023–8654.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Information from this study could be used as an alternative path to induce genetic and epigenetic changes in regenerated plants, which eventually can be used for crop improvement program in eggplant. Moreover, callus induction and its subsequent plant regeneration protocol hold great promise for production of transgenic eggplants of desirable traits by genetic transformation techniques.

Knowledge Management: Journal