The recent advances in genome-wide transcriptome analysis have enabled researchers to identify new genes and manipulate the genome through novel genetic engineering methods to improve plant tolerance to different stresses. We conducted a genome-wide transcriptome analysis to determine tomato genes affecting the salt stress response. The results showed that among the 5784 genes that were responsive to salt stress, 103 genes (1.8%) encode transcription factors, of which 69 (1.2%) were upregulated and 34 (0.6%) were downregulated. The largest group of genes upregulated in response to salt stress (17 genes) is related to the ethylene response transcription factors (ERF) family. Specifically, it was found that the gene JERF1, encoding ERF in tomato, is upregulated in response to salt stress. In the present study we developed a new Agrobacterium-mediated transformation method to deliver the JERF1 gene to mature wheat embryos. It was investigated whether the factors vacuum infiltration, Agrobacterium cell density (OD600 1.0 and its dilutions 1:20, 2:20 and 3:20), the acetosyringone concentration (0, 200 and 400 μM) and the interaction between these factors affect transformation. To this end, Agrobacterium carrying pGWB14-JERF1 was injected into soaked wheat seeds, and T0 transgenic plants were obtained in the greenhouse. Molecular analysis of T0 plants was performed. The highest transformation efficiency was obtained under vacuum infiltration, 200 μM acetosyringone and 1:20 dilution. Notably, we used the in planta transformation method to overcome some of the problems of traditional wheat transformation methods e.g., sterile conditions, recalcitrant regeneration, time-constraints and somaclonal variations.
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This project was funded by Shiraz and Hiroshima Universities. We express appreciation Prof. T Nakagawa, Shimane University, Japan for the kind gift of pGWB14. This project was funded by 82494194.
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Ashrafi-Dehkordi, E., Alemzadeh, A., Tanaka, N. et al. Effects of vacuum infiltration, Agrobacterium cell density and acetosyringone concentration on Agrobacterium-mediated transformation of bread wheat. J Consum Prot Food Saf 16, 59–69 (2021). https://doi.org/10.1007/s00003-020-01312-y
- Agrobacterium tumefaciens
- Microarray analysis
- Vacuum infiltration
- Wheat transformation