Abstract
Plant hormones not only play important roles in regulating plant growth and development, but they also promote cell dedifferentiation and redifferentiation, which play an important role in tissue culture. In the present study, an efficient method of promoting callus plant regeneration for the tissue culture of Brassica juncea (L.) was proposed. We discovered that the callus could be efficiently induced into redifferentiation by 6.8 × 10− 3 µmol/l thidiazuron (TDZ), a cytokinin analogue. To analyze the molecular mechanism of tissue culture, we sequenced the transcriptome of the callus under different culture conditions at two developmental stages. A total of 5362 differentially expressed genes were identified in the TDZ treatments; among them, the expression of 4676 genes was upregulated and that of 686 was downregulated. In the cytokinin signal transduction pathway, seven genes in the Arabidopsis (Arabidopsis thaliana) response regulator (ARR) family were upregulated. In addition, cytokinin affected gene expression in other plant hormone signaling pathways, such as the auxin, ethylene, and brassinosteroid pathways. Most of these genes, such as auxin/indole-3-acetic acid (Aux/IAA) genes and ethylene response factors (ERF-1, ERF-2, and ERF-6), generally showed upregulated expression, which also participated in the regulation of callus plant regeneration. This study provides novel insight into the molecular mechanism of callus plant regeneration in Brassica juncea, which could also provide reference for other plant tissue cultures.
Key message
In Brassica juncea, 6.8 × 10− 3 µmol/l TDZ resulted in the highest regeneration rate of callus, and the genes involved in cytokinin and auxin signal transduction pathways participated in the redifferentiation progress.
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Data availability
The raw data used in this study had been submitted to the Sequence Read Archive (http://www.ncbi.nlm.nih.gov/sra/) under accession PRJNA562990.
Abbreviations
- 2,4-D:
-
2,4-Dichlorophenoxyacetic acid
- NAA:
-
Naphthalene acetic acid
- TDZ:
-
Thidiazuron
- MS:
-
Murashige and Skoog
- CGM:
-
Concentration gradient medium
- H0D15:
-
Callus grown on the CGM containing no hormone for 15 days
- HaD15:
-
Callus grown on the CGM containing only 5.4 × 10−4 µmol/l NAA for 15 days
- HbD15:
-
Callus grown on the CGM containing only 6.8 × 10−3 µmol/l TDZ for 15 days
- H0D21:
-
Callus grown on the CGM containing no hormone for 21 days
- HaD21:
-
Callus grown on the CGM containing only 5.4 × 10−4 µmol/l NAA for 21 days
- HbD21:
-
Callus grown on the CGM containing only 6.8 × 10−3 µmol/l TDZ for 21 days
- T0N0:
-
Callus grown on the CGM containing no hormone
- T0N0.1:
-
Callus grown on the CGM containing only 5.4 × 10−4 µmol/l NAA
- T1.5N0:
-
Callus grown on the CGM containing only 6.8 × 10−3 µmol/l TDZ
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Acknowledgements
This work was financially supported by the National Natural Science Foundation of China (NSFC, Grant No. 31571698); the National Key Research and Development Program of China (Grant No. 2016YFD0101300) and the Program for Modern Agricultural Industrial Technology System of China (Grant No. CARS-12).
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JS and PX conceived and designed the experiments. HL performed the experiments and wrote the manuscript. HL, KH and QX analyzed the data. JW, BY, CM, JT, TF supervised this study. PX and JS edited and revised the manuscript.
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Lu, H., Xu, P., Hu, K. et al. Transcriptome profiling reveals cytokinin promoted callus regeneration in Brassica juncea. Plant Cell Tiss Organ Cult 141, 191–206 (2020). https://doi.org/10.1007/s11240-020-01779-5
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DOI: https://doi.org/10.1007/s11240-020-01779-5