Vol. 72 No: 3

Title:
Improving Micropropagation Conditions for Adult-Phase Shoots of Chestnut

Authors:
M. CONCEPCION SANCHEZ, M. CARMEN SAN-JOSE, E. FERRO, A. BALLESTER and ANA M. VIEITEZ

pp: 433-444

Abstract:
This study aimed to optimize chestnut micropropagation by defining the best explant type for the shoot multiplication stage and by determining a root induction treatment, to balance rooting rate and plantlet quality, and maximize overall survival after transfer to soil. Shoot cultures of Castanea sativa and C. sativa × C. crenata were established in vitro using shoot tips and nodes from shoots forced on stump sprouts, epicormic shoots and crown branches collected from adult trees. The most productive explant type for subculturing was the basal nodal segment, especially when callus tissue was left on the subcultured basal segment. Although rooting capacity was significantly increased when a 7 d 3 mg l^-1 IBA treatment was combined with an initial 5 d dark period, the latter caused severe shoot necrosis. Acceptable rooting frequencies were achieved by a 24 h, 25–50 mg l^-1 IBA treatments without darkness, and for some clones were enhanced by addition of 1% activated charcoal to the root proliferation medium. Generally, high rooting rates were obtained in charcoal medium when material from the base of the trunk had been used as the source of initial explants. In all clones, charcoal improved lateral root development (as reflected in the total root lengths and the projected root areas measured using a digital image analyser), reduced shoot necrosis symptoms and promoted the reinitiation of shoot growth. The rooting rates achieved in a peat/perlite/ vermiculite substrate were similar to those achieved in an charcoal-enriched medium.

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