Abstract
Main conclusion
S. plumbizincicola genetic transformation was optimized using a self-excision molecular-assisted transformation system by integrating the SpGRF4/SpGIF1 gene with XVE and Cre/loxP.
Abstract
Sedum plumbizincicola, despite being an excellent hyperaccumulator of cadmium and zinc with significant potential for soil pollution phytoremediation on farmland, has nonetheless trailed behind other major model plants in genetic transformation technology. In this study, different explants and SpGRF4–SpGIF1 genes were used to optimize the genetic transformation of S. plumbizincicola. We found that petiole and stem segments had higher genetic transformation efficiency than cluster buds. Overexpression of SpGRF4–SpGIF1 could significantly improve the genetic transformation efficiency and shorten the period of obtaining regenerated buds. However, molecular assistance with overexpression of SpGRF4–SpGIF1 leads to abnormal morphology, resulting in plant tissue enlargement and abnormal growth. Therefore, we combined SpGRF4–SpGIF1 with XVE and Cre/loxP to obtain DNA autocleavage transgenic plants induced by estradiol, thereby ensuring normal growth in transgenic plants. This study optimized the S. plumbizincicola genetic transformation system, improved the efficiency of genetic transformation, and established a self-excision molecular-assisted transformation system. This work also established the basis for studying S. plumbizincicola gene function, and for S. plumbizincicola breeding and germplasm innovation.
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Data availability
The Gene and protein sequences of SpGRF4 and SpGIF1 have been uploaded to GenBank, and the submission ID were 2,777,890, 2,777,937. The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- GRF:
-
Growth-regulating factor
- GIF:
-
GRF-interacting factor
- XVE:
-
LexA-VP16-ER
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This research was funded by the National Natural Science Foundation of China (32172666).
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WX and ZS conceived and designed research. YZ, YM, XW, and LH conducted experiments. YZ, HR and WX analyzed data. WX, ZS, and HR supervised the study. YZ wrote the draft of the manuscript. All authors read and approved the manuscript.
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Communicated by Dorothea Bartels.
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Zhang, Y., Mo, Y., Ren, H. et al. Improving Sedum plumbizincicola genetic transformation with the SpGRF4–SpGIF1 gene and the self-excision CRE/LoxP system. Planta 259, 119 (2024). https://doi.org/10.1007/s00425-024-04393-3
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DOI: https://doi.org/10.1007/s00425-024-04393-3