The Optimization of In Vitro Culture Establishment and Shoot Proliferation of “GiSelA 17” (Prunus canescens × Prunus avium): A Novel Cherry Rootstock

“GiSelA 17” (Prunus canescens × Prunus avium) is a novel cherry clonal rootstock with the ability to bear fruit early and resist replant situations, and it has a high tolerance to the menaces of Prunus dwarf virus (PDV) and Prunus necrotic ring spot virus (PNRSV). In this study, two kinds of explants were taken, i.e., shoot tip (E1) (10 mm) and nodal segment (E2) (15 mm) explants. Five different sterilant regimes using sodium hypochlorite, mercuric chloride, and ethyl alcohol were employed to assess surface sterilization. Two types of media, namely Murashige and Skoog (MS) and Woody Plant Medium (WPM), and twelve and six plant growth regulator combinations with benzyl amino purine (BAP) and indole-3-butyric acid (IBA) were used, respectively, for the establishment and proliferation steps. The results show that maximum culture asepsis (75.33%) was obtained with shoot tips (E1) using 0.05% HgCl2 for 5 min + 70% ethanol for 10 s (S4), and maximum explant survival (80.33%) was observed in 0.1% HgCl2 for 5 min (S1) for shoot tips (E1). The maximum establishment rate (83.33%) was found in shoot tips (E1) in MS medium with BAP + IBA (1 + 0.01 mg/L) during the establishment step, with a maximum proliferation rate of 92.00% obtained in MS and BAP (0.75 mg/L). Inferior establishment results (26.66%) were obtained in nodal segments (E2) using WPM and BAP + IBA (1.50 + 0.01 mg/L), with a low proliferation rate (68.66%) in WPM and BAP + IBA (0.25 + 0.01 mg/L). Nonetheless, our research is the first in vitro study on “GiSelA 17” rootstock that focuses on generating the best quality planting material for commercial cherry production.