Abstract
Free asparagine is the precursor for acrylamide, which forms during the baking, toasting and high-temperature processing of foods made from wheat. In this study, CRISPR/Cas9 was used to knock out the asparagine synthetase gene, TaASN2, of wheat (Triticum aestivum) cv. Cadenza. A 4-gRNA polycistronic gene was introduced into wheat embryos by particle bombardment and plants were regenerated. T1 plants derived from 11 of 14 T0 plants were shown to carry edits. Most edits were deletions (up to 173 base pairs), but there were also some single base pair insertions and substitutions. Editing continued beyond the T1 generation. Free asparagine concentrations in the grain of plants carrying edits in all six TaASN2 alleles (both alleles in each genome) were substantially reduced compared with wildtype, with one plant showing a more than 90 % reduction in the T2 seeds. A plant containing edits only in the A genome alleles showed a smaller reduction in free asparagine concentration in the grain, but the concentration was still lower than in wildtype. Free asparagine concentration in the edited plants was also reduced as a proportion of the free amino acid pool. Free asparagine concentration in the T3 seeds remained substantially lower in the edited lines than wildtype, although it was higher than in the T2 seeds, possibly due to stress. In contrast, the concentrations of free glutamine, glutamate and aspartate were all higher in the edited lines than wildtype. Low asparagine seeds showed poor germination but this could be overcome by exogenous application of asparagine.
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Claims 12
Claim · model-written summary
T1 descendants of 11 of 14 T0 plants carried edits.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- carry editse0
- Value
- 11 of 14e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“T1 plants derived from 11 of 14 T0 plants were shown to carry edits.”
t0“wheat”
Claim · model-written summary
Edits included deletions up to 173 base pairs.
- Taxon
- wheatTriticum aestivumt0
- Gene
- TaASN2t0
- Outcome
- deletionse0
- Value
- up to 173 base pairse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Most edits were deletions (up to 173 base pairs), but there were also some single base pair insertions and substitutions.”
t0“wheat with greatly reduced accumulation of free asparagine in the grain, produced by CRISPR/Cas9 editing of asparagine synthetase gene TaASN2”
Claim · model-written summary
Single base pair insertions and substitutions also occurred.
- Taxon
- wheatTriticum aestivumt0
- Outcome
- single base pair insertions and substitutionse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“but there were also some single base pair insertions and substitutions.”
t0“wheat”
Claim · model-written summary
Editing persisted beyond T1.
- Taxon
- wheatTriticum aestivumt0
- Outcome
- Editing continuede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Editing continued beyond the T1 generation.”
t0“wheat”
Claim · model-written summary
Editing all six TaASN2 alleles substantially reduced grain free asparagine versus wild type.
- Taxon
- wheatTriticum aestivumt0
- Treatment
- edits in all six TaASN2 allelese0
- Gene
- TaASN2e0
- Outcome
- Free asparagine concentrations in the graine0
- Direction
- reducede0
- Value
- more than 90 %e0
- Comparator
- wildtypee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Free asparagine concentrations in the grain of plants carrying edits in all six TaASN2 alleles (both alleles in each genome) were substantially reduced compared with wildtype, with one plant showing a more than 90 % reduction in the T2 seeds.”
t0“wheat”
Claim · model-written summary
A genome allele edits produced a smaller grain asparagine reduction than all-allele edits, but remained below wild type.
- Taxon
- wheatTriticum aestivumt0
- Treatment
- edits only in the A genome allelese0
- Outcome
- free asparagine concentration in the graine0
- Direction
- lowere0
- Comparator
- wildtypee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A plant containing edits only in the A genome alleles showed a smaller reduction in free asparagine concentration in the grain, but the concentration was still lower than in wildtype.”
t0“wheat”
Claim · model-written summary
The free asparagine share of the free amino acid pool declined in edited plants.
- Taxon
- wheatTriticum aestivumt0
- Treatment
- edited plantse0
- Outcome
- Free asparagine concentratione0
- Direction
- reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Free asparagine concentration in the edited plants was also reduced as a proportion of the free amino acid pool.”
t0“wheat”
Claim · model-written summary
T3 seed free asparagine remained below wild type.
- Taxon
- wheatTriticum aestivumt0
- Treatment
- edited linese0
- Outcome
- Free asparagine concentration in the T3 seedse0
- Direction
- lowere0
- Comparator
- wildtypee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Free asparagine concentration in the T3 seeds remained substantially lower in the edited lines than wildtype, although it was higher than in the T2 seeds, possibly due to stress.”
t0“wheat”
Claim · model-written summary
T3 seed free asparagine exceeded T2 levels.
- Taxon
- wheatTriticum aestivumt1
- Outcome
- free asparaginet0
- Direction
- highere0
- Comparator
- T2 seedse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“although it was higher than in the T2 seeds, possibly due to stress.”
t0“free asparagine in the grain”
t1“wheat”
Claim · model-written summary
Free glutamine, glutamate and aspartate were higher in edited lines.
- Taxon
- wheatTriticum aestivumt0
- Treatment
- edited linese0
- Outcome
- concentrations of free glutamine, glutamate and aspartatee0
- Direction
- highere0
- Comparator
- wildtypee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the concentrations of free glutamine, glutamate and aspartate were all higher in the edited lines than wildtype.”
t0“wheat”
Claim · model-written summary
Low-asparagine seeds germinated poorly.
- Taxon
- wheatTriticum aestivumt0
- Material
- Low asparagine seedse0
- Outcome
- poor germinatione0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Low asparagine seeds showed poor germination”
t0“wheat”
Claim · model-written summary
Exogenous asparagine overcame poor germination of low-asparagine seeds.
- Taxon
- wheatTriticum aestivumt0
- Treatment
- exogenous application of asparaginee0
- Outcome
- poor germinatione0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Low asparagine seeds showed poor germination but this could be overcome by exogenous application of asparagine.”
t0“wheat”
The same questions in other papers
asparagine · growth · Triticum aestivum (wheat)Results agree
1 paper in the same direction as this paper
- positive
loss of function · molecular: metabolites · Triticum aestivum (wheat)Positive and negative results
13 papers in the same direction as this paper · 18 in the opposite direction
- negative
- no effect
- negative
28 papers more in this group
See all 32 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.