New methods for sorghum transformation in temperate climates

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Sorghum (Sorghum bicolor) is an emerging cereal crop in temperate climates due to its high drought tolerance and other valuable traits. Genetic transformation is an important tool for the improvement of cereals. However, sorghum is recalcitrant to genetic transformation which is almost only successful in warmer climates. Here, we test the application of two new techniques for sorghum transformation in temperate climates, namely transient transformation by Agrobacterium tumefaciens–mediated agroinfiltration and stable transformation using gold particle bombardment and leaf whorls as explants. We optimized the transient transformation method, including post-infiltration incubation of plants in the dark and using Agrobacterium grown on plates with a high cell density (OD600 = 2.0). Expression of the green fluorescence protein (GFP)-tagged endogenous sorghum gene SbDHR2 was achieved with low transformation efficiency, and our results point out a potential weakness in using this approach for localization studies. Furthermore, we succeeded in the production of callus and somatic embryos from leaf whorls, although no genetic transformation was accomplished with this method. Both methods show potential, even if they seem to be influenced by climatic conditions and therefore need further optimization to be applied routinely in temperate climates.

OriginalsprogEngelsk
Artikelnummerplad030
TidsskriftAoB PLANTS
Vol/bind15
Udgave nummer3
Antal sider14
ISSN2041-2851
DOI
StatusUdgivet - 2023

Bibliografisk note

Funding Information:
The research was funded by VILLUM FONDEN, Denmark, grant no. 19151, awarded to N.B., and the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 801199, awarded to S.M. Acknowledgements

Funding Information:
The research was funded by VILLUM FONDEN, Denmark, grant no. 19151, awarded to N.B., and the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 801199, awarded to S.M. We thank Dr. Luo Guangbin and Professor Michael Palmgren (University of Copenhagen, Denmark) for access to and instructions in use of the gene canon, Professor Ida Elisabeth Johansen (University of Copenhagen, Denmark) for advice concerning plasmid cloning and tissue culture and Dr. Karen Massel (University of Queensland, Australia) for providing the plasmid for particle bombardment.

Publisher Copyright:
© The Author(s) 2023.

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