From induction to embryo proliferation: improved somatic embryogenesis protocol in grapevine for Italian cultivars and hybrid Vitis rootstocks

Abstract Somatic embryogenesis is the most common regeneration method for application of new genomic techniques like cisgenesis/intragenesis, genome editing, and RNAi. However, some local important genotypes show recalcitrance to this morphogenetic strategy, which represents an obstacle genetic engineering techniques. Whole flowers, stamens, pistils three different Italian Vitis vinifera L. cultivars (Ancellotta, Glera, Lambrusco Salamino), four hybrid rootstocks (110 Richter, 17.37, SO4, Star 50) have been tested in several culture media with changing basal salts (NN MS), combinations growth regulators (BAP, 2,4-D, NOA, PIC, NAA), gelling agents, initiate somatic embryogenesis. The formation embryogenic calli was observed mainly from whole flowers cultured on PIV medium salts, B5 vitamins, 3 g L −1 gelrite, 60 sucrose, 8.9 µM BAP, 4.5 2,4-D), stamens MS1 (MS 7 plant agar, 20 5 cv. Ancellotta, Salamino, all rootstocks, except 50, showed best embryogenetic response placed MS1. In a recalcitrant as MS supplemented 1 gelrite agent, highest percentage addition, two-step protocol efficiently optimized further induction secondary embryo production above-listed grapevine genotypes, guaranteed long-term maintenance cultures clusters or single embryos.