Efficient CRISPR/Cas9-mediated genome editing in Rehmannia glutinosa
| Autor: | Xinrong Li, Feng-Qing Wang, Jingyu Zhi, Mingming Li, Xu Yang, Xin Zuo, Hongzheng Sun, Cai-Xia Xie, Zhongyi Zhang |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Chlorophyll
0106 biological sciences 0301 basic medicine Phytoene desaturase Agrobacterium Plant Science 01 natural sciences Marker gene 03 medical and health sciences Genome editing Gene Expression Regulation Plant CRISPR Cloning Molecular Gene Plant Proteins Gene Editing Genetics biology Cas9 fungi food and beverages General Medicine Plants Genetically Modified Rehmannia glutinosa biology.organism_classification Carotenoids Rehmannia 030104 developmental biology Mutation CRISPR-Cas Systems Oxidoreductases Agronomy and Crop Science Genome Plant 010606 plant biology & botany |
| Zdroj: | Plant Cell Reports. 40:1695-1707 |
| ISSN: | 1432-203X 0721-7714 |
| Popis: | Here, we cloned a phytoene desaturase (PDS) gene from Rehmannia glutinosa, and realized RgPDS1 knock out in R. glutinosa resulted in the generation of albino plants. Rehmannia glutinosa is a highly important traditional Chinese medicine (TCM) with specific pharmacology and economic value. R. glutinosa is a tetraploid plant, to date, no report has been published on gene editing of R. glutinosa. In this study, we combined the transcriptome database of R. glutinosa and the reported phytoene desaturase (PDS) gene sequences to obtain the PDS gene of R. glutinosa. Then, the PDS gene was used as a marker gene to verify the applicability and gene editing efficiency of the CRISPR/Cas9 system in R. glutinosa. The constructed CRISPR/Cas9 system was mediated by Agrobacterium to genetically transform into R. glutinosa, and successfully regenerated fully albino and chimeric albino plants. The next-generation sequencing (NGS) confirmed that the albino phenotype was indeed caused by RgPDS gene target site editing, and it was found that base deletion was more common than insertion or replacement. Our results revealed that zCas9 has a high editing efficiency on the R. glutinosa genome. This research lays a foundation for further use of gene editing technology to study the molecular functions of genes, create excellent germplasm, accelerate domestication, and improve the yield and quality of R. glutinosa. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink