Shoot regeneration from various explants of horse chestnut (Aesculus hippocastanum L.)

• Micropropagation protocol via adventitious shoot regeneration in horse chestnut (Aesculus hippocastanum) was established.
• Adventitious shoot formation was observed in all types of explants (leaf and stem).
• Regeneration frequency and shoot number depended on explant type and growth regulators.
• TDZ has lower effect on shoot regeneration in stem and leaf explants compared to BA.
• Shoot regeneration from in vitro leaf explants in horse chestnut is reported for the first time.

A protocol is described for the adventitious shoot bud regeneration from leaf and stem explants in Aesculus hippocastanum. Effect of different media (Heller, WPM, Rugini and DKW) and growth regulators (BA, TDZ and NAA) combinations on shoot organogenesis were examined. Culture medium content had a significant effect on shoot regeneration in Aesculus. The highest shoot number and length was achieved on Rugini and WPM medium. Adventitious shoot formation was observed in all types of explants. Regeneration frequency and shoot number depended on explant type and growth regulators. The highest regeneration (100%) and the highest shoot bud number (9.7) were recorded in stem explants on WPM medium with 8.9 μM BA + 0.5 μM NAA. Generally, TDZ was less effective on shoot regeneration in stem and leaf explants compared to BA. The addition of NAA to the culture medium improved shoot regeneration from stem discs. Rooting of elongated shoots was significantly increased (68.1%) on full-strength WPM medium supplemented with 2.7 μM NAA. Rooted plantlets were acclimatized successfully under greenhouse conditions.