Journal of Pure and Applied MicrobiologyVol. 9 No. Special Edition Nov. 2015

Efficient Microspore Embryogenesis Induction in Tomato (Lycopersicon esculentum Mill.) using Shed Microspore Culture

Behzad Ahmadi1,2, Mehran E. Shariatpanahi2*, Rasoul Asghari-Zakaria1, Nasser Zare1 and Pejman Azadi2

1Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Mohaghegh Ardabili, Ardabil, Iran. 2Department of Tissue Culture and Gene Transformation, Agricultural Biotechnology Research Institute of Iran (ABRII), AREEO, 3135933151 Karaj, Iran.

Received on 13 March 2015 and accepted on 15 April 2015

 

ABSTRACT

The effects of cold treatment (4?C for 0, 2, and 5 days), heat shock (30?C for 0, 2, and 5 days), and two induction media (B5 and NLN) supplemented with 2% and 9% sucrose were assessed on microspore embryogenesis induction in tomato using shed microspore culture technique. Significantly higher callusing was obtained when anthers were cultured on the B5 medium with 2.0 mg l-1 2, 4-D and 2.0 mg l-1 BAP. In addition, IAA at 0.5 mg l-1 in combination with 0.5 mg l-1 Kin also substantially improved callogenesis. Chitosan treatment proved to be beneficial to callusing and shoot regeneration so that, high callogenesis (85%) and shoot regeneration (3%) was obtained from anthers treated with 50 mg l-1 chitosan. Microspore-derived structures with more than 9 nuclei were observed in both media containing 2% sucrose when treated at 4?C for 2 days and 30?C for 2 and 5 days. Microspore-derived embryos (MDEs) were only obtained in the double-layer B5 medium supplemented with 2% sucrose which exposed to 4?C for 2 days and then 30?C for 2 and 5 days. Microspore embryogenesis in tomato could be efficiently induced when appropriate duration of cold treatment, heat shock, and induction medium were selected.

Keywords : Lycopersicon esculentum Mill.; Microspore embryogenesis; Shed microspore culture