Abstract
Late blight, caused by Phytophthora infestans, is severely damaging to the global tomato industry. Micro-RNAs (miRNAs) have been widely demonstrated to play vital roles in plant resistance by repressing their target genes. Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) method has been continuously improved and extensively applied to edit plant genomes. However, editing multiplex miRNAs by CRISPR/Cas9 in tomato has not been studied yet. We knocked out miR482b and miR482c simultaneously in tomato through the multiplex CRISPR/Cas9 system. Two transgenic plants with silenced miR482b and miR482c simultaneously and one transgenic line with silenced miR482b alone were obtained. Compared with wild-type plants, the disease symptoms of three transgenic plants upon infection were reduced, accompanied by increased expression of their common target nucleotide binding site-leucine-rich repeat genes and decreased levels of reactive oxygen species. Furthermore, silencing miR482b and miR482c simultaneously was more resistant than silencing miR482b alone in tomato. More importantly, we found that knocking out miR482b and miR482c can elicit expression perturbation of other miRNAs, suggesting cross-regulation between miRNAs. Our study demonstrated that editing miR482b and miR482c simultaneously with CRISPR/Cas9 is an efficient strategy for generating pathogen-resistant tomatoes, and cross-regulation between miRNAs may reveal the novel mechanism in tomato–P. infestans interactions.
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Claims 7
Claim · model-written summary
Multiplex editing produced two lines with both miRNAs silenced and one with miR482b silenced.
- Taxon
- tomato
Solanum lycopersicumt0 - Gene
- miR482b and miR482ce0
- Outcome
- transgenic plants with silenced miR482b and miR482ce0
- Value
- Two transgenic plantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Two transgenic plants with silenced miR482b and miR482c simultaneously and one transgenic line with silenced miR482b alone were obtained.”
t0“Tomato”
Claim · model-written summary
The three edited plants had reduced disease symptoms after infection compared with wild type.
- Taxon
- tomatoSolanum lycopersicumt0
- Treatment
- three transgenic plantse0
- Interactor
- Phytophthora infestans
Phytophthora infestanst0pathogen - Outcome
- disease symptomse0
- Direction
- reducede0
- Comparator
- wild-type plantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Compared with wild-type plants, the disease symptoms of three transgenic plants upon infection were reduced”
t0“Tomato Resistance toPhytophthora infestans”
Claim · model-written summary
The edited plants had increased expression of common target NBS-LRR genes.
- Taxon
- tomatoSolanum lycopersicumt0
- Treatment
- transgenic plantse1
- Gene
- nucleotide binding site-leucine-rich repeat genese0
- Outcome
- expression of their common target nucleotide binding site-leucine-rich repeat genese0
- Direction
- increasede0
- Comparator
- wild-type plantse2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“accompanied by increased expression of their common target nucleotide binding site-leucine-rich repeat genes”
t0“Editing miR482b and miR482c Simultaneously by CRISPR/Cas9 Enhanced Tomato Resistance toPhytophthora infestans”
e1“Two transgenic plants with silenced miR482b and miR482c simultaneously and one transgenic line with silenced miR482b alone were obtained.”
e2“Compared with wild-type plants, the disease symptoms of three transgenic plants upon infection were reduced”
Claim · model-written summary
Edited plants had decreased reactive oxygen species levels.
- Taxon
- tomatoSolanum lycopersicumt0
- Treatment
- transgenic plantse1
- Outcome
- levels of reactive oxygen speciese0
- Direction
- decreasede0
- Comparator
- wild-type plantse2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“decreased levels of reactive oxygen species”
t0“Editing miR482b and miR482c Simultaneously by CRISPR/Cas9 Enhanced Tomato Resistance toPhytophthora infestans”
e1“Two transgenic plants with silenced miR482b and miR482c simultaneously and one transgenic line with silenced miR482b alone were obtained.”
e2“Compared with wild-type plants, the disease symptoms of three transgenic plants upon infection were reduced”
Claim · model-written summary
Double silencing provided greater resistance than miR482b silencing alone.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- silencing miR482b and miR482c simultaneouslye0
- Interactor
- Phytophthora infestansPhytophthora infestanst0pathogen
- Gene
- miR482b and miR482ce0
- Outcome
- resistante0
- Direction
- more resistante0
- Comparator
- silencing miR482b alonee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“silencing miR482b and miR482c simultaneously was more resistant than silencing miR482b alone in tomato”
t0“Phytophthora infestans”
Claim · model-written summary
Knocking out miR482b and miR482c perturbed expression of other miRNAs.
- Taxon
- tomatoSolanum lycopersicumt0
- Treatment
- knocking out miR482b and miR482ce0
- Gene
- miR482b and miR482ce0
- Outcome
- expression perturbation of other miRNAse0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“knocking out miR482b and miR482c can elicit expression perturbation of other miRNAs”
t0“Tomato”
Claim · model-written summary
The authors propose simultaneous CRISPR editing as a strategy to generate resistant tomatoes.
- Taxon
- tomatoesSolanum lycopersicume0
- Gene
- miR482b and miR482ce0
- Outcome
- efficient strategy for generating pathogen-resistant tomatoese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“editing miR482b and miR482c simultaneously with CRISPR/Cas9 is an efficient strategy for generating pathogen-resistant tomatoes”
The same questions in other papers
loss of function · Phytophthora infestans · Solanum lycopersicum (tomato)Positive and negative results
9 papers in the same direction as this paper · 2 in the opposite direction
- negative
- positive
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8 papers more in this group
See all 12 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.