Abstract
Dof (DNA binding with one finger) proteins are plant-specific transcription factors involved in gene regulation during abiotic and biotic stresses. Our previous studies suggested that tobacco Dof proteins, including BBF3, can upregulate the expression of the virus resistance N gene as well as resistance-related genes. In this study, we generated and characterized transgenic tobacco lines carrying the N gene that overexpressed BBF3 cDNA constitutively. In the absence of virus challenge, the BBF3 overexpression caused no developmental defects and no changes in the N transcript level, although the transcript levels of some defense-related genes were affected compared with levels in non-transgenic plants. TMV infection induced a hypersensitive reaction (HR) with larger necrotic lesions in transgenic lines than in non-transgenic plants, but there was no change in total virus accumulation per lesion in a standardized area. Activation of the N gene expression in non-transgenic plants during synchronously-induced HR was followed by downregulation of BBF3 gene expression. Constitutive overexpression of the BBF3 transgene resulted in higher accumulation of the N gene as well as HR marker genes (Hin1 and Hsr203j) and lower accumulation of salicylic acid- and jasmonic acid-signaling marker genes (PR1-a and PR-1b). In particular, the expression of PR-1b was remarkably suppressed in the BBF3-overexpressing transgenic lines. Dof-binding motifs were confirmed in the upstream regions of all target genes examined. Our results suggest that BBF3 has a role in modulating the expression of genes associated with virus elicitor recognition and defense responses resulting in larger areas of HR-associated cell death.
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Acknowledgements
We also thank the Institute of Global Innovation Research at Tokyo University of Agriculture and Technology (GIR-TUAT) for the Special Research Fund. This work was supported partly by the Japan Society for Promotion of Science (JSPS) KAKENHI Grant Numbers 26450052 and 19K06047 (to N.S.).
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Japan Society for the Promotion of Science, 26450052, Nobumitsu SASAKI, 19K06047, Nobumitsu SASAKI.
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NS, YM, RSN conceived and designed the study. TS, MF, MS, and KS prepared materials and performed the experiments. TS, MF, RSN, and NS analyzed the data. TS, MF, and NS wrote the manuscript with edits and additional text from RSN and YM.
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Suzuki, T., Fujita, M., Suzuki, M. et al. Overexpression of BBF3 encoding a tobacco Dof transcription factor alters gene expression and cell death induction during N-mediated hypersensitivity to tobacco mosaic virus. J Gen Plant Pathol 90, 2–13 (2024). https://doi.org/10.1007/s10327-023-01155-z
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DOI: https://doi.org/10.1007/s10327-023-01155-z