Full Length Research Paper

Somatic embryogenesis and plant regeneration of recalcitrant cottons (Gossypium hirsutum)

G. Y. Han, X. F. Wang, G. Y. Zhang and Z. Y. Ma*

Key Laboratory of Crop Germplasm Resources of Hebei, Agricultural University of Hebei, Baoding 071001, People’s Republic of China.

*Corresponding author. E-mail: mzhy@hebau.edu.cn. Tel: +86-312-7528401. Fax: +86-312-7528400.

Accepted 3 October, 2008

Many restrictive factors still remain in cotton tissue culture such as long duration, unpredictability and a high degree of genotype dependence. The main objective of this study was to develop a protocol allowing consistent somatic embryogenesis and plant regeneration from five recalcitrant cotton cultivars. Our results showed that the best medium for calli induction is MSB (MS medium + vitamine B5) supplemented with indolebutyric acid (IBA, 0.1 mg.l^-1), kinetin (KIN, 0.1 mg.l^-1) and 2,4-dichlorophenoxyacetic acid (2,4-D, 0.1 mg.l^-1). Embryogenic calli of all the five genotypes used were successfully from MSB medium supplemented with IBA (0.3 mg.l^-1) and KIN (0.05 mg.l^-1). Somatic embryos and transformation of somatic embryos into plants were successfully induced on MSB medium supplemented with ½×NH₄NO₃ (825 mg.l^-1), 2×KNO₃ (3800 mg.l^-1), glutamine (2.0 g/l) and asparagines (0.5 g/l). The protocol developed in this study for cotton plant regeneration could be shortened to 4 - 5 months. Furthermore, the firstly-obtained regenerated plants of above five cultivars will broaden the range of genotypes for in vitro manipulation for cotton improvement.

Key words: Cotton, callus induction, somatic embryogenesis, embryogenic callus, plant regeneration.