An efficient in vitro shoot regeneration method from leaf explants of apple cultivars Golden Delicious and Melrose by optimization of regeneration medium, explant type and orientation, dark pre-treatment, and gelling agent is presented. Murashige and Skoog’s regeneration medium containing 22 μM thidiazuron (TDZ) and 1.5 μM indole-3-butyric acid (IBA) (M2 medium) was superior for regeneration as well as for subsequent shoot multiplication in both cultivars, providing regeneration frequency of 95% or higher in the best combination with other factors. Pre-incubation in the dark proved to be an essential factor for regeneration. The use of agar as a gelling agent provides satisfactory regeneration frequency compared with media gelled with Phytagel TM . Leaf explants of cv. Melrose with adaxial surface in contact with M2 medium and those of cv. Golden Delicious orientated contrary regenerated the highest mean number of shoots per explant. Under optimal conditions, a maximal index of shoot-forming capacity of 11.44 and 6.30 for ‘Melrose’ and ‘Golden Delicious’, respectively, was achieved. Regenerated shoots were successfully rooted and acclimated ex vitro.
Seed treatments with zinc, boron, biostimulant Coveron and MIX (zinc + boron + Coveron) were applied to three lettuce and three celeriac cultivars. Seeds of three wheat cultivars were treated under laboratory conditions with Trichoderma harzianum and eight Bacillus spp. Seed germination, seedling growth, and the presence of the following pathogens were determined: Fusarium sp., Alternaria sp., Penicillium sp., and Mucor sp. The Coveron treatment was the most effective on lettuce seeds tested in the germination cabinet. Seed germination was higher by 4% than in the control. Alternatively, germination of seeds treated with boron in the greenhouse was higher by 12% than in the control. The Coveron treatment had the highest effect on the shoot length, which was greater by 0.7 and 2.1 cm in the germination cabinet and the greenhouse, respectively. This treatment was also the most effective on the root length. Zn, B, and MIX treatments increased celeriac seed germination by 14% in the germination cabinet. The Zn treatment was the most efficient on seeds tested in the greenhouse. The germination was higher by 15%. A significant cultivar × treatment interaction was determined in both observed species under both conditions. The maximum effect on wheat seed germination (8%) was achieved with the T. harzianum treatment in the Salazar cultivar. A significant interdependence (p ≤ 0.01 to p ≤ 0.001) was established between seed germination and the seedling growth. The interrelationship between seed germination and pathogens of all cultivars was negative.