The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of <i>Triticum timopheevii</i>
<i>Triticum timopheevii</i> Zhuk. is a tetraploid wheat that is utilized worldwide as a valuable breeding source for wheat improvement. Gene-based biotechnologies can contribute to this field; however, <i>T. timopheevii</i> exhibits recalcitrance and albinism in tissue cultur...
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MDPI AG
2021-11-01
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| Series: | Plants |
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| author | Dmitry Miroshnichenko Anna Klementyeva Sergey Dolgov |
| author_facet | Dmitry Miroshnichenko Anna Klementyeva Sergey Dolgov |
| author_sort | Dmitry Miroshnichenko |
| collection | DOAJ |
| description | <i>Triticum timopheevii</i> Zhuk. is a tetraploid wheat that is utilized worldwide as a valuable breeding source for wheat improvement. Gene-based biotechnologies can contribute to this field; however, <i>T. timopheevii</i> exhibits recalcitrance and albinism in tissue cultures, making this species of little use for manipulation through genetic engineering and genome editing. This study tested various approaches to increasing in vitro somatic embryogenesis and plant regeneration, while reducing the portion of albinos in cultures derived from immature embryos (IEs) of <i>T. timopheevii</i>. They included (i) adjusting the balance between 2,4-D and daminozide in callus induction medium; (ii) cultivation using various darkness/illumination schedules; and (iii) inclusion of additional concentrations of copper ions in the tissue culture medium. We achieved a 2.5-fold increase in somatic embryogenesis (up to 80%) when 50 mg L<sup>−1</sup> daminozide was included in the callus induction medium together with 3 mg L<sup>−1</sup> 2,4-D. It was found that the dark cultivation for 20–30 days was superior in terms of achieving maximum culture efficiency; moreover, switching to light in under 2 weeks from culture initiation significantly increased the number of albino plants, suppressed somatic embryogenesis, and decreased the regeneration of green plants. Media containing higher levels of copper ions did not have a positive effect on the regeneration of green plants; contrarily, the elevated concentrations caused albinism in plantlets. The results and relevant conclusions of the present study might be valuable for establishing an improved protocol for the regeneration of green plants in tissue cultures of <i>T. timopheevii</i>. |
| first_indexed | 2024-03-10T03:17:16Z |
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| language | English |
| last_indexed | 2024-03-10T03:17:16Z |
| publishDate | 2021-11-01 |
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| series | Plants |
| spelling | doaj.art-2f97a4ba41e64e7eb64b1d21d4f1060b2023-11-23T10:10:46ZengMDPI AGPlants2223-77472021-11-011012262010.3390/plants10122620The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of <i>Triticum timopheevii</i>Dmitry Miroshnichenko0Anna Klementyeva1Sergey Dolgov2Kurchatov Genomics Center of All-Russia Research Institute of Agricultural Biotechnology, Timiryazevskaya Street 42, 127550 Moscow, RussiaBranch of Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russion Academy of Science, 142290 Pushchino, RussiaBranch of Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russion Academy of Science, 142290 Pushchino, Russia<i>Triticum timopheevii</i> Zhuk. is a tetraploid wheat that is utilized worldwide as a valuable breeding source for wheat improvement. Gene-based biotechnologies can contribute to this field; however, <i>T. timopheevii</i> exhibits recalcitrance and albinism in tissue cultures, making this species of little use for manipulation through genetic engineering and genome editing. This study tested various approaches to increasing in vitro somatic embryogenesis and plant regeneration, while reducing the portion of albinos in cultures derived from immature embryos (IEs) of <i>T. timopheevii</i>. They included (i) adjusting the balance between 2,4-D and daminozide in callus induction medium; (ii) cultivation using various darkness/illumination schedules; and (iii) inclusion of additional concentrations of copper ions in the tissue culture medium. We achieved a 2.5-fold increase in somatic embryogenesis (up to 80%) when 50 mg L<sup>−1</sup> daminozide was included in the callus induction medium together with 3 mg L<sup>−1</sup> 2,4-D. It was found that the dark cultivation for 20–30 days was superior in terms of achieving maximum culture efficiency; moreover, switching to light in under 2 weeks from culture initiation significantly increased the number of albino plants, suppressed somatic embryogenesis, and decreased the regeneration of green plants. Media containing higher levels of copper ions did not have a positive effect on the regeneration of green plants; contrarily, the elevated concentrations caused albinism in plantlets. The results and relevant conclusions of the present study might be valuable for establishing an improved protocol for the regeneration of green plants in tissue cultures of <i>T. timopheevii</i>.https://www.mdpi.com/2223-7747/10/12/2620wheatimmature embryossomatic embryogenesisalbino phenotype2,4-DCuSO<sub>4</sub> |
| spellingShingle | Dmitry Miroshnichenko Anna Klementyeva Sergey Dolgov The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of <i>Triticum timopheevii</i> Plants wheat immature embryos somatic embryogenesis albino phenotype 2,4-D CuSO<sub>4</sub> |
| title | The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of <i>Triticum timopheevii</i> |
| title_full | The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of <i>Triticum timopheevii</i> |
| title_fullStr | The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of <i>Triticum timopheevii</i> |
| title_full_unstemmed | The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of <i>Triticum timopheevii</i> |
| title_short | The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of <i>Triticum timopheevii</i> |
| title_sort | effect of daminozide dark light schedule and copper sulphate in tissue culture of i triticum timopheevii i |
| topic | wheat immature embryos somatic embryogenesis albino phenotype 2,4-D CuSO<sub>4</sub> |
| url | https://www.mdpi.com/2223-7747/10/12/2620 |
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