Abstract
Genetic modification of rice is mainly carried out by Agrobacterium-mediated transformation of callus accompanied by tissue culture. It is time consuming, laborious and unapplicable for cultivars unable to induce callus. In this study, we have reported a novel gene transfer protocol that involves pulling out primary leaf from coleoptile and injection of Agrobacterium culture into the empty channel. Out of 25 plants survived after injection of Agrobacterium tumefaciens EHA105 culture harboring pCAMBIA1301-RD29A-AtDREB1A, 8 T0 plants revealed the expected size of around 811 bp corresponding to AtDREB1A gene and Southern blotting analysis on 18 T1 plants suggested introgression of AtDREB1A. 3 T2 lines (7–9, 12–3, 18–6) exhibited accumulation of free proline and soluble sugars, yet increase of chlorophyll content, but decrease of electrolyte leakage and methane dicarboxylic aldehyde under cold stress condition at the vegetative growth stage. Yield components investigation on T2 lines showed earlier heading date and no yield loss compared to wild type plants grown under normal condition. GUS expression analysis and integrated transgene detection in T0 and T1 plants followed by evaluation of cold stress tolerance in T2 lines suggest the advantage of this in planta transformation protocol to obtain transgenic rice.
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Abbreviations
- AtDREB :
-
Arabidopsis thaliana dehydration responsive element binding
- hpt :
-
Hygromycin phosphotransferase
- GUS:
-
Beta-glucuronidase
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Acknowledgements
We thank all members who helped with vector construction, microscope sample preparation and photographing, plant regeneration and field works. This research was made possible by financial support of the Scientific and Technical Commission, D P R Korea.
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UH and YC conceptualized and designed the study, UH and SS conducted the experiments, and MP and MK constructed the vectors, KK, TH and YC analysed the data; UH and SS collected data and wrote the first draft; and all authors contributed substantially to revisions.
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Ho, UH., Song, SR., Choe, YI. et al. Development of Agrobacterium-mediated in planta transformation protocol through coleoptile in rice. Transgenic Res 32, 339–349 (2023). https://doi.org/10.1007/s11248-023-00355-9
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DOI: https://doi.org/10.1007/s11248-023-00355-9