New methods for sorghum transformation in temperate climates
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New methods for sorghum transformation in temperate climates. / Miller, Sara; Rønager, Asta; Holm, Rose; Fontanet-Manzaneque, Juan B.; Caño-Delgado, Ana I.; Bjarnholt, Nanna.
In: AoB PLANTS, Vol. 15, No. 3, plad030, 2023.Research output: Contribution to journal › Review › Research › peer-review
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TY - JOUR
T1 - New methods for sorghum transformation in temperate climates
AU - Miller, Sara
AU - Rønager, Asta
AU - Holm, Rose
AU - Fontanet-Manzaneque, Juan B.
AU - Caño-Delgado, Ana I.
AU - Bjarnholt, Nanna
N1 - Publisher Copyright: © The Author(s) 2023.
PY - 2023
Y1 - 2023
N2 - 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.
AB - 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.
KW - Agrobacterium
KW - dhurrinase
KW - sorghum
KW - tissue culture
KW - transformation
KW - transient
U2 - 10.1093/aobpla/plad030
DO - 10.1093/aobpla/plad030
M3 - Review
C2 - 37396498
AN - SCOPUS:85164823504
VL - 15
JO - AoB PLANTS
JF - AoB PLANTS
SN - 2041-2851
IS - 3
M1 - plad030
ER -
ID: 360690780