Abstract
Key message
To establish a sterile culture system and protoplast regeneration system for Bryum argenteum, and to establish and apply CRISPR/Cas9 system in Bryum argenteum.
Abstract
Bryum argenteum is a fascinating, cosmopolitan, and versatile moss species that thrives in various disturbed environments. Because of its comprehensive tolerance to the desiccation, high UV and extreme temperatures, it is emerging as a model moss for studying the molecular mechanisms underlying plant responses to abiotic stresses. However, the lack of basic tools such as gene transformation and targeted genome modification has hindered the understanding of the molecular mechanisms underlying the survival of B. argenteum in different environments. Here, we reported the protonema of B. argenteum can survive up to 95.4% water loss. In addition, the genome size of B. argenteum is approximately 313 Mb by kmer analysis, which is smaller than the previously reported 700 Mb. We also developed a simple method for protonema induction and an efficient protoplast isolation and regeneration protocol for B. argenteum. Furthermore, we established a PEG-mediated protoplast transient transfection and stable transformation system for B. argenteum. Two homologues of ABI3(ABA-INSENSITIVE 3) gene were successfully cloned from B. argenteum. To further investigate the function of the ABI3 gene in B. argenteum, we used the CRISPR/Cas9 genetic editing system to target the BaABI3A and BaABI3B gene in B. argenteum protoplasts. This resulted in mutagenesis at the target in about 2–5% of the regenerated plants. The isolated abi3a and abi3b mutants exhibited increased sensitivity to desiccation, suggesting that BaABI3A and BaABI3B play redundant roles in desiccation stress. Overall, our results provide a rapid and simple approach for molecular genetics in B. argenteum. This study contributes to a better understanding of the molecular mechanisms of plant adaptation to extreme environmental.
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Data availability
Raw data of B. argenteum CNU51 resequencing were uploaded to the National Genomics Data Center (https://ngdc.cncb.ac.cn/) in the sample accession SAMC2820342 the project ID PRJCA017895.
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Acknowledgements
We thank Fabien Nogué (Institut Jean-Pierre Bourgin, National Institute for Agricultural Research, France) for providing the pAct–Cas9 and the pBNRF plasmid.
Funding
This work was supported by grants from the Major Research Plan of National Nature Science Foundation of China (91631109 to Y.He), the General Program of National Natural Science Foundation of China (No.31970658 Y.He. and No.30971558 to Y.Hu.).
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YH, FB and YH conceived and designed the project. GZ, RS FL, and YH performed most of experiments and analyzed the data. Other authors assisted in experiments and discussed the results. FL and YH wrote the manuscript.
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Leng, F., Zhou, G., Shi, R. et al. Development of PEG-mediated genetic transformation and gene editing system of Bryum argenteum as an abiotic stress tolerance model plant. Plant Cell Rep 43, 63 (2024). https://doi.org/10.1007/s00299-024-03143-9
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DOI: https://doi.org/10.1007/s00299-024-03143-9