Chestnuts gall wasp (Dryocosmus kuriphilus Yasumatsu) is threatening the existence of the European chestnut (Castanea sativa Mill.) in Slovenia and in Europe as a whole. The main goal of the assignment was to, via in vitro techniques, propagate and root two chestnut genes sources marked P. and Š2 and prevent their collapse. The chestnut gene source for material was harvested from trees growing in the Vipava Valley. MS-½NO3 medium supplemented with BAP (0.5 mg/L) was used to reproduce the shoots, which proved to be very successful in both genotypes. The propagation was followed by the rooting phase on MS-½NO3 medium with different concentrations of auxins IBA, NAA or IAA and additives: sugar (sucrose, glucose and fructose) and coconut water. The responsiveness of the shoots to rooting was tested with fifteen culture media and none proved successful. In addition to rooting, the vitality of shoots was monitored on selected media: the number of decayed, vitrified, with dead tips, and shoots with and without callus. After the statistical analysis of the data, we found no significant differences between the two gene sources in the observed traits, but there were significant differences between the media. The highest number of failed shoots was on K4 medium (54 %). K11 had the highest number of shoots with dead tips (31 %). Medium K15 had the highest number of vitrified shoots (33 %). Medium K5 formed the most callus shoots (23 %). We found that the species and concentration of auxin have the greatest influence on the vitality of the shoots.
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