This study was conducted to investigate the efficacy of shoot regeneration from different leaf types (normal leaves and vitrified leaves) from three different carnation cultivars ‘Kumbuyl’, ‘Denev’, and ‘Jinju’ using different combinations of 3-indole butyric acid (IBA) and thidiazuron (TDZ) concentrations. The shoot tips cultured on Murashige and Skoog (MS) basal media (Type 1 media) produced normal leaves, while those cultured-on media supplemented with plant growth regulators and/or vitamin (Type 2 media and Type 3 media) produced vitrified leaves for all cultivars. Culture of normal leaf segments on MS medium containing different combinations of IBA and TDZ concentrations induced callus in all treatments; however, the callus was unable to induce shoots and finally became necrotic. In contrast, no callus induction was observed in the control (hormone-free treatment). When vitrified leaf segments underwent the same treatments, shoots were induced from the vitrified leaves (derived from Type 2 media) but were unhealthy and gradually died, whereas those induced from Type 3 media were vitrified and healthy. The optimal combination for the best shoot regeneration and number of shoots per explants varied depending on the genotypes used. The vitrified shoots induced from the leaves of Type 3 media transformed into normal shoots and survived well under greenhouse conditions. According to the results of random amplified polymorphic DNA (RAPD) analysis, the banding patterns of twelve primers that were detected in vitrified leaf-induced normalized shoots were identical to those of normal in vitro grown plants, indicating that no genetic variation had occurred during the procedure. Taken together, this study indicates that vitrified leaves can be used for shoot regeneration of recalcitrant carnation cultivars, regardless of the genotypes and types of vitrified leaves. However, as the number of shoots per explants was still low, further investigation is warranted to obtain a more efficient shoot regeneration protocol for genetic transformation of the cultivars.

中文翻译：

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从不同康乃馨品种的体外玻璃化叶片中再生遗传稳定的植物。

这项研究旨在研究使用3种吲哚丁酸（IBA）的不同组合来自三种不同康乃馨品种'Kumbuyl'，'Denev'和'Jinju'的不同叶片类型（正常叶片和玻璃化叶片）的再生芽的功效）和噻唑隆（TDZ）浓度。在Murashige和Skoog（MS）基础培养基（1型培养基）上培养的芽尖可产生正常叶片，而在补充有植物生长调节剂和/或维生素的培养基（2型培养基和3型培养基）上培养的茎尖则可制成玻璃化叶片。所有品种。在所有处理中，在含有IBA和TDZ浓度不同组合的MS培养基上培养正常叶片片段会诱导愈伤组织；然而，愈伤组织不能诱导芽并最终坏死。相反，在对照中（无激素处理）未观察到愈伤组织的诱导。当对玻璃化叶段进行相同处理时，从玻璃化叶中诱导出芽（衍生自2型培养基），但不健康并逐渐死亡，而从3型培养基中诱导出的芽则被玻璃化且健康。最佳芽再生和每外植体芽数的最佳组合取决于所使用的基因型。从3型培养基的叶片诱导的玻璃化芽转化为正常芽，并在温室条件下存活良好。根据随机扩增多态性DNA（RAPD）分析的结果，在玻璃化叶片诱导的标准化芽中检测到的十二种引物的条带模式与正常体外生长的植物相同，表明在手术过程中没有发生遗传变异。两者合计，这项研究表明玻璃化的叶可用于顽固康乃馨品种的芽再生，而与玻璃化叶的基因型和类型无关。但是，由于每个外植体的芽数仍然很少，因此有必要进行进一步的研究以获得更有效的芽再生方案，以进行品种的遗传转化。