The present study reports an efficient in vitro plant regeneration system for amphidiploid (2n = 42) spelt wheat (Triticum spelta L.). Plant regeneration from mature embryos of winter spelt variety “Europe” was carried out as a two-step process. The first step was callus induction on a medium supplemented with 2 mg L⁻¹ dichlorophenoxyacetic acid and 10 mg L⁻¹ silver nitrate resulting in 96% callus formation. The second step resulted in 30% plant regeneration frequency from calluses transferred to Murashige and Skoog medium supplemented with 2 mg L⁻¹ 6-benzylaminopurine, 0.5 mg L⁻¹ 1-naphthaleneacetic acid, and 10 mg L⁻¹ silver nitrate. Regeneration medium supplemented with 1 mg L⁻¹ 6-benzylaminopurine, 0.2 mg L⁻¹ picloram, and the hormone-free medium turned out to be less effective in our experiments. Regenerated plants formed roots after transfer to half-strength Murashige and Skoog medium supplemented with 0.7 mg L⁻¹ indole-3-butyric acid. About one-third of the resulting regenerated plants that formed roots were transferred to soil. The inter-simple sequence repeat markers were used to confirm the genetic homogeneity of regenerated plants. Thus, our regeneration protocol can be used for in vitro regeneration of spelt plants from mature embryos with minimal risk of genetic variability and provides an essential step towards developing an efficient strategy for spelt genetic transformation.

本研究报告了一种有效的体外植物再生系统，用于二倍体（2 n  = 42）种小麦（Triticum spelta L.）。从冬季拼写的“欧洲”品种的成熟胚中再生植物的过程分为两个步骤。第一步是在补充有2 mg L ^-1二氯苯氧基乙酸和10 mg L ^-1硝酸银的培养基上诱导愈伤组织，从而形成96％的愈伤组织。第二步使从愈伤组织转移到补充2 mg L ^-1 6-苄基氨基嘌呤，0.5 mg L ^-1 1-萘乙酸和10 mg L ^-1的Murashige和Skoog培养基中的植物再生频率达到30％硝酸银。在我们的实验中，补充有1 mg L ^-1 6-苄氨基嘌呤，0.2 mg L ^-1吡咯烷和无激素培养基的再生培养基效果较差。再生植株转移到补充有0.7 mg L ^-1吲哚-3-丁酸的半强度Murashige和Skoog培养基后形成根。约有三分之一形成根的再生植物转移到土壤中。简单序列间重复标记用于确认再生植物的遗传同质性。因此，我们的再生方案可用于体外 从成熟胚中再生出咒语植物，遗传变异风险极小，这为制定咒语遗传转化的有效策略提供了必不可少的步骤。