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In vitro plant regeneration from mature embryos of amphidiploid spelt Triticum spelta L.

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Abstract

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.

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Acknowledgements

The seed material was provided by LLC “Ukrainian Scientific Institute of Breeding” (Ukraine). The work was performed within the fundamental departmental topics of the Department of Genetic Engineering Institute of Cell Biology and Genetic Engineering of NAS of Ukraine: III-8-17 “Study of the functioning of heterologous genes and their impact on the adaptive characteristics of plant systems under biotic and abiotic stresses” (№ state registration 0117U002589, 2017–2021).

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Correspondence to A.V. Kyriienko.

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Editor: Tom Clemente

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Kyriienko, A., Shcherbak, N.L., Kuchuk, M.V. et al. In vitro plant regeneration from mature embryos of amphidiploid spelt Triticum spelta L.. In Vitro Cell.Dev.Biol.-Plant 57, 856–863 (2021). https://doi.org/10.1007/s11627-021-10158-4

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