Data underlying the publication: Development of high frequency in vitro shoot regeneration system from leaves of apple cultivar Oregon Spur and optimization of antibiotics concentration

the present study is carried out optimize the high efficiency shoot regeneration protocol from the leaf explant of the Oregon Spur cultivar. The different concentration of the Cytokinin (BAP, TDZ) and auxin (NAA, IBA) were tested to identify the optimium ratio and quantity required to generate shoots. 4mg/l BA alongwith 0.2mg/l NAA was found more effective cytokinin than TDZ for high quality shoot induction. Effect of dark treatment was also tested on the shoot regeneration and inhbitory effect was observed. During optimization of the antibiotics concentration, Leaf explant were found highly sensitive to Kanamycin and inhibited shoot induction above 5mg/l, whereas cefotaxime at 100-300mg/l promoted organogenesis. 500mg/l cefotaxime was found optimal to eliminate bacterial overgrowth after co-cultivation and 5 mg/l of kanamycin was found best for selection of plant.

本研究旨在优化俄勒冈斯普尔（Oregon Spur）品种叶片外植体（explant）的高效不定芽再生体系。本研究设置了不同浓度梯度的细胞分裂素（Cytokinin，含BAP、TDZ）与生长素（Auxin，含NAA、IBA）处理，以筛选出诱导不定芽所需的最佳配比与用量。结果表明，相较于TDZ，4mg/L BAP与0.2mg/L NAA联用可更高效地诱导出高质量不定芽。本研究还测试了暗处理对不定芽再生的影响，观察到其具有显著抑制效应。在抗生素浓度优化实验中，叶片外植体对卡那霉素（Kanamycin）敏感性极高，当浓度超过5mg/L时会抑制不定芽诱导；而100~300mg/L的头孢噻肟（cefotaxime）可促进器官发生。研究发现，500mg/L头孢噻肟为共培养（co-cultivation）后抑制细菌过度增殖的最佳浓度，而5mg/L卡那霉素则为植物筛选的最适浓度。