Abstract
Abstract Broomrapes ( Phelipanche aegyptiaca and Orobanche spp.) are obligate plant parasites that cause extreme damage to crop plants. The parasite seeds have strict requirements for germination, involving preconditioning and exposure to specific chemicals strigolactones [SLs] exuded by the host roots. SLs are plant hormones derived from plant carotenoids via a pathway involving the Carotenoid Cleavage Dioxygenase 8 ( CCD8) . Having no effective means to control parasitic weeds in most crops, and with CRISPR/Cas9 being an effective gene-editing tool, here we demonstrate that CRISPR/Cas9-mediated mutagenesis of the CCD8 gene can be used to develop host resistance to the parasitic weed P . aegyptiaca . Cas9/single guide (sg) RNA constructs were targeted to the second exon of CCD8 in tomato ( Solanum lycopersicum L .) plants. Several CCD8 Cas9 mutated tomato lines with variable insertions or deletions in CCD8 were obtained with no identified off-targets. Genotype analysis of T1 plants showed that the introduced CCD8 mutations are inherited. Compared to control tomato plants, the CCD8 Cas9 mutant had morphological changes that included dwarfing, excessive shoot branching and adventitious root formation. In addition, SL-deficient CCD8 Cas9 mutants showed a significant reduction in parasite infestation compared to non-mutated tomato plants. In the CCD8 Cas9 mutated lines, orobanchol (SL) content was significantly reduced but total carotenoids level and expression of genes related to carotenoid biosynthesis were increased, as compared to control plants. Taking into account, the impact of plant parasitic weeds on agriculture and difficulty to constitute efficient control methods, the current study offers insights into the development of a new, efficient method that could be combined with various collections of resistant tomato rootstocks.
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Claims 10
Claim · model-written summary
CCD8 Cas9 mutation produced tomato lines with variable insertions or deletions and no identified off-targets.
- Taxon
- tomato
Solanum lycopersicume0 - Gene
- CCD8e0
- Outcome
- variable insertions or deletions in CCD8e0
- Comparator
- no identified off-targetse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Several CCD8 Cas9 mutated tomato lines with variable insertions or deletions in CCD8 were obtained with no identified off-targets.”
Claim · model-written summary
The introduced CCD8 mutations were inherited in T1 plants.
- Taxon
- tomatoSolanum lycopersicumt0
- Organ / stage
- T1 plantse0
- Gene
- CCD8e0
- Outcome
- introduced CCD8 mutations are inheritede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Genotype analysis of T1 plants showed that the introduced CCD8 mutations are inherited.”
t0“tomato”
Claim · model-written summary
The CCD8 Cas9 tomato mutant was dwarfed relative to control tomato plants.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- CCD8 Cas9 mutante0
- Gene
- CCD8e0
- Outcome
- dwarfinge0
- Comparator
- control tomato plantse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Compared to control tomato plants, the CCD8 Cas9 mutant had morphological changes that included dwarfing, excessive shoot branching and adventitious root formation.”
Claim · model-written summary
The CCD8 Cas9 tomato mutant had excessive shoot branching relative to controls.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- CCD8 Cas9 mutante0
- Gene
- CCD8e0
- Outcome
- excessive shoot branchinge0
- Comparator
- control tomato plantse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Compared to control tomato plants, the CCD8 Cas9 mutant had morphological changes that included dwarfing, excessive shoot branching and adventitious root formation.”
Claim · model-written summary
The CCD8 Cas9 tomato mutant formed adventitious roots relative to controls.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- CCD8 Cas9 mutante0
- Gene
- CCD8e0
- Outcome
- adventitious root formatione0
- Comparator
- control tomato plantse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Compared to control tomato plants, the CCD8 Cas9 mutant had morphological changes that included dwarfing, excessive shoot branching and adventitious root formation.”
Claim · model-written summary
SL-deficient CCD8 Cas9 mutants had significantly reduced parasite infestation relative to non-mutated tomato plants.
- Taxon
- tomatoSolanum lycopersicume0
- Treatment
- SL-deficient CCD8 Cas9 mutantse0
- Interactor
- Phelipanche aegyptiaca
Phelipanche aegyptiacat0parasite - Gene
- CCD8e0
- Outcome
- parasite infestatione0
- Direction
- reductione0
- Comparator
- non-mutated tomato plantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“SL-deficient CCD8 Cas9 mutants showed a significant reduction in parasite infestation compared to non-mutated tomato plants.”
t0“Phelipanche aegyptiaca”
Claim · model-written summary
Orobanchol content was significantly reduced in CCD8 Cas9 mutated tomato lines versus control plants.
- Taxon
- tomatoSolanum lycopersicumt0
- Treatment
- CCD8 Cas9 mutated linese0
- Gene
- CCD8e0
- Outcome
- orobanchol (SL) contente0
- Direction
- reducede0
- Comparator
- control plantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“In the CCD8 Cas9 mutated lines, orobanchol (SL) content was significantly reduced but total carotenoids level and expression of genes related to carotenoid biosynthesis were increased, as compared to control plants.”
t0“tomato”
Claim · model-written summary
Total carotenoid level increased in CCD8 Cas9 mutated tomato lines versus control plants.
- Taxon
- tomatoSolanum lycopersicumt0
- Treatment
- CCD8 Cas9 mutated linese0
- Gene
- CCD8e0
- Outcome
- total carotenoids levele0
- Direction
- increasede0
- Comparator
- control plantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“In the CCD8 Cas9 mutated lines, orobanchol (SL) content was significantly reduced but total carotenoids level and expression of genes related to carotenoid biosynthesis were increased, as compared to control plants.”
t0“tomato”
Claim · model-written summary
Expression of carotenoid biosynthesis genes increased in CCD8 Cas9 mutated tomato lines versus controls.
- Taxon
- tomatoSolanum lycopersicumt0
- Treatment
- CCD8 Cas9 mutated linese0
- Gene
- CCD8e0
- Outcome
- expression of genes related to carotenoid biosynthesise0
- Direction
- increasede0
- Comparator
- control plantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“In the CCD8 Cas9 mutated lines, orobanchol (SL) content was significantly reduced but total carotenoids level and expression of genes related to carotenoid biosynthesis were increased, as compared to control plants.”
t0“tomato”
Claim · model-written summary
The authors suggest combining the editing approach with resistant tomato rootstock collections.
- Taxon
- tomatoSolanum lycopersicume0
- Outcome
- combined with various collections of resistant tomato rootstockse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the current study offers insights into the development of a new, efficient method that could be combined with various collections of resistant tomato rootstocks.”
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See all 80 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.