Abstract
BACKGROUND: Rice leaf blight, which is a devastating disease worldwide, is caused by the bacterium Xanthomonas oryzae pv. oryzae (Xoo). The upregulated by transcription activator-like 1 (UPT) effector box in the promoter region of the rice Xa13 gene plays a key role in Xoo pathogenicity. Mutation of a key bacterial protein-binding site in the UPT box of Xa13 to abolish PXO99-induced Xa13 expression is a way to improve rice resistance to bacteria. Highly efficient generation and selection of transgene-free edited plants are helpful to shorten and simplify the gene editing-based breeding process. Selective elimination of transgenic pollen of T0 plants can enrich the proportion of T1 transgene-free offspring, and expression of a color marker gene in seeds makes the selection of T2 plants very convenient and efficient. In this study, a genome editing and multiplexed selection system was used to generate bacterial leaf blight-resistant and transgene-free rice plants. RESULTS: We introduced site-specific mutations into the UPT box using CRISPR/Cas12a technology to hamper with transcription-activator-like effector (TAL) protein binding and gene activation and generated genome-edited rice with improved bacterial blight resistance. Transgenic pollen of T0 plants was eliminated by pollen-specific expression of the α-amylase gene Zmaa1, and the proportion of transgene-free plants increased from 25 to 50% among single T-DNA insertion events in the T1 generation. Transgenic seeds were visually identified and discarded by specific aleuronic expression of DsRed, which reduced the cost by 50% and led to up to 98.64% accuracy for the selection of transgene-free edited plants. CONCLUSION: We demonstrated that core nucleotide deletion in the UPT box of the Xa13 promoter conferred resistance to rice blight, and selection of transgene-free plants was boosted by introducing multiplexed selection. The combination of genome editing and transgene-free selection is an efficient strategy to accelerate functional genomic research and plant breeding.
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Claims 5
Claim · model-written summary
CRISPR/Cas12a mutations in the UPT box generated rice with improved bacterial blight resistance.
- Taxon
- rice
Oryza sativae0 - Treatment
- site-specific mutations into the UPT boxe0
- Interactor
- Xanthomonas oryzae pv. oryzae
Xanthomonas oryzae pv. oryzaee2pathogen - Gene
- Xa13e1
- Outcome
- bacterial blight resistancee0
- Direction
- improvede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We introduced site-specific mutations into the UPT box using CRISPR/Cas12a technology to hamper with transcription-activator-like effector (TAL) protein binding and gene activation and generated genome-edited rice with improved bacterial blight resistance”
e1“core nucleotide deletion in the UPT box of the Xa13 promoter conferred resistance to rice blight”
e2“Xanthomonas oryzae pv. oryzae (Xoo)”
t0“rice”
Claim · model-written summary
Pollen-specific Zmaa1 expression increased the transgene-free share of T1 offspring from 25 to 50%.
- Taxon
- riceOryza sativat0
- Treatment
- pollen-specific expression of the α-amylase gene Zmaa1e0
- Gene
- Zmaa1e0
- Outcome
- proportion of transgene-free plantse0
- Direction
- increasede0
- Value
- 25 to 50%e0
- Qualifier
- among single T-DNA insertion events in the T1 generatione0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic pollen of T0 plants was eliminated by pollen-specific expression of the α-amylase gene Zmaa1, and the proportion of transgene-free plants increased from 25 to 50% among single T-DNA insertion events in the T1 generation”
t0“rice”
Claim · model-written summary
Aleurone-specific DsRed marking reduced the cost of selecting transgene-free edited rice by 50%.
- Taxon
- riceOryza sativat0
- Treatment
- specific aleuronic expression of DsRede0
- Gene
- DsRede0
- Outcome
- coste0
- Direction
- reducede0
- Value
- 50%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transgenic seeds were visually identified and discarded by specific aleuronic expression of DsRed, which reduced the cost by 50% and led to up to 98.64% accuracy for the selection of transgene-free edited plants”
t0“rice”
Claim · model-written summary
Aleurone-specific DsRed marking yielded up to 98.64% accuracy in selecting transgene-free edited rice.
- Taxon
- riceOryza sativat0
- Treatment
- specific aleuronic expression of DsRede0
- Gene
- DsRede0
- Outcome
- accuracy for the selection of transgene-free edited plantse0
- Value
- up to 98.64%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“specific aleuronic expression of DsRed, which reduced the cost by 50% and led to up to 98.64% accuracy for the selection of transgene-free edited plants”
t0“rice”
Claim · model-written summary
Genome editing combined with transgene-free selection was presented as an efficient strategy to accelerate functional genomics and plant breeding.
- Taxon
- riceOryza sativat0
- Treatment
- combination of genome editing and transgene-free selectione0
- Outcome
- accelerate functional genomic research and plant breedinge0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The combination of genome editing and transgene-free selection is an efficient strategy to accelerate functional genomic research and plant breeding”
t0“rice”
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