Abstract

To establish an effective protocol for plant regeneration through indirect organogenesis, effects of explants type, culture media and plant growth regulators on callus induction and shoot regeneration of chestnut (Castanea sativa Mill.) were investigated. Three different explants (root, nodal and internodal segment), two different media [Murashige and Skoog medium (MS) and Gamborg's B5 (B5)] and different plant growth regulators (6-benzylaminopurine (BA), thidiazuron (TDZ), indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA)) with different concentration (0.2, 0.5, 1 and 1.5 mgLˉ¹) for shoot and root induction were chosen. The results show that nodal segment was the best explant for callus induction (69.4%) when cultured on MS medium supplemented with 1 mgLˉ¹ TDZ and MS was the best medium to induce callus formation (74.6%). The highest shoot multiplication (66.9%) was observed on MS medium with 0.2 mgLˉ¹ TDZ. Regenerated shoots were rooted in vitro on MS containing 1.5 mgLˉ¹ IBA. Also, plantlets with well developed root and shoot systems were acclimatized inside the green house and 80% of the plantlets survived on transfer to garden soil. This protocol provides a basis for future studies on genetic improvement.

Key words: Chestnut, node, internode segment, indirect organogenesis, callus formation, shoot regeneration.

Abbreviation

Abbreviations: BA, 6-Benzylaminopurine; IAA, indole-3-acetic acid; TDZ, thidiazuron; IBA, indole-3-butyric acid; MS, Murashige and Skoog medium; B₅, Gamborg's B₅.