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Living Modified Organism
(LMO)
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Spring barley with a mutation in the ARE1 gene (GP-koARE-S1-2-A)
EN
GP-koARE-S1-2-A
No
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Klopina,Organization:Usovsko ()Private sector (business and industry)
78973, CzechiaPhone: +420 583 484 111,Fax:Email: klopina@usovsko.cz,Website: http://usovsko.cz/?p=o-nas,
Spring barley (Hordeum vulgare L.) was modified to alter nitrogen-use efficiency and leaf senescence through targeted mutagenesis of the endogenous abnormal reproductive organ development 1 (ARE1) gene. The ARE1 gene encodes a regulator involved in nitrogen metabolism, nutrient remobilization and the progression of leaf senescence. To achieve this modification, a 15 bp deletion together with a single nucleotide insertion was introduced into the coding sequence of the endogenous barley ARE1 gene, generating a loss-of-function allele. The mutation disrupts the normal reading frame of the gene and introduces a premature termination codon, resulting in the loss of functional ARE1 protein activity. Disruption of ARE1 function has been associated with altered nitrogen utilization, delayed senescence and improved nitrogen-use efficiency under nitrogen-limiting conditions.
The term “Recipient organism” refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas “Parental organisms” refers to those that were involved in cross breeding or cell fusion.
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BCH-ORGA-SCBD-12110-5 Organism Hordeum vulgare (Barley, HORVU)Crops
Spring barley (Hordeum vulgare L.) cultivar Golden Promise
EN
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Spring barley with a mutation in the ARE1 gene (GP-koARE-S1-3-A)| Usovsko | Changes in physiology and/or production, Tolerance to abiotic stress (Nitrogen deficiency)
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Spring barley with a mutation in the ARE1 gene (GP-koARE-S2-1-A)| Usovsko | Changes in physiology and/or production, Tolerance to abiotic stress (Nitrogen deficiency)
CPGE_VEC00113
EN
- Agrobacterium-mediated DNA transfer
- Gene editing (e.g. CRISPR-Cas, etc.)
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
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BCH-GENE-SCBD-299474-1 ARE1 | Hordeum vulgare (Barley, HORVU)Protein coding sequence | Changes in physiology and/or production
Information on the inserted DNA sequence
The modified barley was generated using CRISPR/Cas-mediated genome editing following transformation with vector CPGE_VEC00113. The editing construct was subsequently removed through segregation, resulting in plants containing the intended mutation in the endogenous ARE1 locus and no retained vector-derived sequences.
The endogenous ARE1 coding sequence is 1,278 bp in length. In line GP-koARE-S1-2-A, the modified ARE1 allele contains a 15-bp deletion in the first exon, located 60 bp downstream of the start codon, resulting in the deletion of five amino acids near the N-terminus of the protein. The allele also contains a single-nucleotide (A) insertion 109 bp downstream of the start codon. This insertion disrupts the reading frame and introduces a premature stop codon, preventing production of a functional ARE1 protein and resulting in a loss-of-function allele. The resulting modified gene is 1,264 bp in length. The modified sequence is derived entirely from the endogenous barley ARE1 locus and contains no heterologous genetic material or novel coding sequence.
ARE1 functions in the regulation of nitrogen metabolism, nutrient remobilization and leaf senescence. Loss of ARE1 function has been associated with enhanced nitrogen-use efficiency and altered senescence-related physiological processes in barley.
Note
- Molecular characterization confirmed the intended mutations within the endogenous ARE1 locus.
- Molecular analyses confirmed the absence of vector-derived transgenic sequences in the final selected generation following segregation.
- No foreign DNA or novel coding sequence is present in the final GP-koARE-S1-2-A line.
EN
- Food
- Research
EN
EN
| Record type | Field | Record(s) | |
|---|---|---|---|
| Risk Assessment generated by a regulatory process | Living modified organism(s) | 1 | |
| Living Modified Organism | Related LMO(s) | 2 | |
