Abstract
Summary CRISPR /Cas9 has been widely used for genome editing in many organisms, including important crops like wheat. Despite the tractability in designing CRISPR /Cas9, efficacy in the application of this powerful genome editing tool also depends on DNA delivery methods. In wheat, the biolistics based transformation is the most used method for delivery of the CRISPR /Cas9 complex. Due to the high frequency of gene silencing associated with co‐transferred plasmid backbone and low edit rate in wheat, a large T 0 transgenic plant population are required for recovery of desired mutations, which poses a bottleneck for many genome editing projects. Here, we report an Agrobacterium ‐delivered CRISPR /Cas9 system in wheat, which includes a wheat codon optimized Cas9 driven by a maize ubiquitin gene promoter and a guide RNA cassette driven by wheat U6 promoters in a single binary vector. Using this CRISPR /Cas9 system, we have developed 68 edit mutants for four grain‐regulatory genes, Ta CKX 2‐1 , Ta GLW 7 , Ta GW 2, and Ta GW 8 , in T 0 , T 1 , and T 2 generation plants at an average edit rate of 10% without detecting off‐target mutations in the most Cas9‐active plants. Homozygous mutations can be recovered from a large population in a single generation. Different from most plant species, deletions over 10 bp are the dominant mutation types in wheat. Plants homozygous of 1160‐bp deletion in Ta CKX 2‐D1 significantly increased grain number per spikelet. In conclusion, our Agrobacterium ‐delivered CRISPR /Cas9 system provides an alternative option for wheat genome editing, which requires a small number of transformation events because CRISPR /Cas9 remains active for novel mutations through generations.
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Claims 5
Claim · model-written summary
The Agrobacterium-delivered CRISPR/Cas9 system gave an average edit rate of 10% in wheat.
- Taxon
- wheat
Triticum aestivumt0 - Outcome
- edit ratee2
- Value
- 10%e2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“for wheat genome editing”
e2“at an average edit rate of 10%”
Claim · model-written summary
The system yielded 68 edit mutants in wheat.
- Taxon
- wheatTriticum aestivumt0
- Outcome
- edit mutantse1
- Value
- 68e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“for wheat genome editing”
e1“Using this CRISPR /Cas9 system, we have developed 68 edit mutants”
Claim · model-written summary
Homozygous mutations can be recovered from a large wheat population in a single generation.
- Taxon
- wheatTriticum aestivumt0
- Outcome
- Homozygous mutationse3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“for wheat genome editing”
e3“Homozygous mutations can be recovered from a large population in a single generation”
Claim · model-written summary
In wheat, deletions over 10 bp are the dominant mutation types.
- Taxon
- wheatTriticum aestivumt0
- Outcome
- dominant mutation typese4
- Qualifier
- deletions over 10 bpe4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“for wheat genome editing”
e4“Different from most plant species, deletions over 10 bp are the dominant mutation types in wheat”
Claim · model-written summary
Homozygous 1160‐bp deletion in Ta CKX 2‐D1 significantly increased grain number per spikelet in wheat.
- Taxon
- wheatTriticum aestivume6
- Treatment
- 1160‐bp deletion in Ta CKX 2‐D1e5
- Gene
- Ta CKX 2‐D1e5
- Outcome
- grain number per spikelete5
- Direction
- increasede5
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“for wheat genome editing”
e5“Plants homozygous of 1160‐bp deletion in Ta CKX 2‐D1 significantly increased grain number per spikelet”
e6“important crops like wheat”
The same questions in other papers
loss of function · yield · Triticum aestivum (wheat)Positive and negative results
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- positive
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