ArticlePlant physiology2024
Efficient gene editing of a model fern species through gametophyte-based transformation.
Article in Plant physiology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- Sulfur metabolism-dependent retrograde signaling for oxidative stress acclimation.Journal of experimental botany · 2026Review
- Article
- Sex-type specification and meristem development in fern gametophytes.Discover developmental biology · 2026Review
- Non-seed plant research in the spotlight.Biology open · 2025Article
- Molecular Evolution of Plant SULTR Proteins and Expression Analysis of HvSULTR Under Heat Stress in Barley.Plants (Basel, Switzerland) · 2025Article
- Resolving the Stasis-Dynamism Paradox: Genome Evolution in Tree Ferns.Molecular biology and evolution · 2025Article
- Analysis of auxin responses in the fern Ceratopteris richardii identifies the developmental phase as a major determinant for response properties.Development (Cambridge, England) · 2024Article
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Authors and funding
13 authors.
Funding
Abstract
The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease (Cas) system allows precise and easy editing of genes in many plant species. However, this system has not yet been applied to any fern species through gametophytes due to the complex characteristics of fern genomes, genetics, and physiology. Here, we established a protocol for gametophyte-based screening of single-guide RNAs (sgRNAs) with high efficiency for CRISPR/Cas9-mediated gene knockout in a model fern species, Ceratopteris richardii. We utilized the C. richardii ACTIN promoter to drive sgRNA expression and the enhanced CaMV 35S promoter to drive the expression of Streptococcus pyogenes Cas9 in this CRISPR-mediated editing system, which was employed to successfully edit a few genes, such as Nucleotidase/phosphatase 1 (CrSAL1) and Phytoene Desaturase (CrPDS), which resulted in an albino phenotype in C. richardii. Knockout of CrSAL1 resulted in significantly (P < 0.05) reduced stomatal conductance (gs), leaf transpiration rate (E), guard cell length, and abscisic acid (ABA)-induced reactive oxygen species (ROS) accumulation in guard cells. Moreover, CrSAL1 overexpressing plants showed significantly increased net photosynthetic rate (A), gs, and E as well as most of the stomatal traits and ABA-induced ROS production in guard cells compared to the wild-type (WT) plants. Taken together, our optimized CRISPR/Cas9 system provides a useful tool for functional genomics in a model fern species, allowing the exploration of fern gene functions for evolutionary biology, herbal medicine discovery, and agricultural applications.
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