Selection and evaluation of gene-edited knockout mutants of AtAAP2 and AtCRF4 homologs of rice for agronomic nitrogen use efficiency (ANUE)
Nitrogen (N) is essential for amino acid synthesis in rice production, but its excessive use poses an environmental concern. This research aimed to improve rice agronomic nitrogen use efficiency (ANUE) by knockout (KO) of rice homologs of the two selected genes from Arabidopsis thaliana: AtAAP2, an...
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| Format: | Online |
| Language: | eng |
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Sociedad Colombiana de Ciencias Hortícolas-SCCH and Universidad Pedagógica y Tecnológica de Colombia-UPTC
2023
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| Online Access: | https://revistas.uptc.edu.co/index.php/ciencias_horticolas/article/view/16120 |
| _version_ | 1801706310455001088 |
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| author | Wakatabi, Kentaro Selvaraj, Michael Gomez Guzmán-Prada, Diego Alexander Cuásquer, Juan Bosco López-López, Karina Endo, Masaki Ishitani, Manabu |
| author_facet | Wakatabi, Kentaro Selvaraj, Michael Gomez Guzmán-Prada, Diego Alexander Cuásquer, Juan Bosco López-López, Karina Endo, Masaki Ishitani, Manabu |
| author_sort | Wakatabi, Kentaro |
| collection | OJS |
| description | Nitrogen (N) is essential for amino acid synthesis in rice production, but its excessive use poses an environmental concern. This research aimed to improve rice agronomic nitrogen use efficiency (ANUE) by knockout (KO) of rice homologs of the two selected genes from Arabidopsis thaliana: AtAAP2, an amino acid permease involved in N transportation in shoots, and AtCRF4, a transcription factor participating in N uptake in roots. The homologs of these genes in rice were identified based on amino acid sequence similarity and knocked out using CRISPR/Cas9 mediated gene editing (GE). The AAP2-KO and CRF4-KO lines were subjected to agronomic evaluations with three N doses: 100% (180 kg ha-1), 50% (90 kg ha-1), and 0% (0 kg ha-1) and showed a 130-175% increase in dry biomass weight and a 183-313% increase in panicle number compared to wild type (WT) in the first experiment. These lines also had slower leaf senescence, the so-called “stay-green” trait, indicating the KO effect of target genes in N metabolism. However, neither AAP2-KO nor CRF4-KO showed better yield or ANUE than WT. This study demonstrated the usefulness of GE technology in gene evaluation and highlighted the effects of AtAAP2 and AtCRF4 genes in the plant N cycle. |
| format | Online |
| id | oai:oai.revistas.uptc.edu.co:article-16120 |
| institution | Revista Colombiana de Ciencias Hortícolas |
| language | eng |
| publishDate | 2023 |
| publisher | Sociedad Colombiana de Ciencias Hortícolas-SCCH and Universidad Pedagógica y Tecnológica de Colombia-UPTC |
| record_format | ojs |
| spelling | oai:oai.revistas.uptc.edu.co:article-161202024-02-14T21:28:32Z Selection and evaluation of gene-edited knockout mutants of AtAAP2 and AtCRF4 homologs of rice for agronomic nitrogen use efficiency (ANUE) Selección y evaluación de mutantes knockout genéticamente editados de homólogos de AtAAP2 y AtCRF4 de arroz para la eficiencia agronómica del uso del nitrógeno (ANUE) Wakatabi, Kentaro Selvaraj, Michael Gomez Guzmán-Prada, Diego Alexander Cuásquer, Juan Bosco López-López, Karina Endo, Masaki Ishitani, Manabu Amino acid permease 2 (AAP2) Cytokinin response factor 4 (CRF4) Oryza sativa L. Remote sensing Rice Nitrogen Aminoácido permeasa 2 (AAP2) Factor de respuesta de citoquinina 4 (CRF4) Oryza sativa L. Sensores remotos Arroz Nitrógeno Nitrogen (N) is essential for amino acid synthesis in rice production, but its excessive use poses an environmental concern. This research aimed to improve rice agronomic nitrogen use efficiency (ANUE) by knockout (KO) of rice homologs of the two selected genes from Arabidopsis thaliana: AtAAP2, an amino acid permease involved in N transportation in shoots, and AtCRF4, a transcription factor participating in N uptake in roots. The homologs of these genes in rice were identified based on amino acid sequence similarity and knocked out using CRISPR/Cas9 mediated gene editing (GE). The AAP2-KO and CRF4-KO lines were subjected to agronomic evaluations with three N doses: 100% (180 kg ha-1), 50% (90 kg ha-1), and 0% (0 kg ha-1) and showed a 130-175% increase in dry biomass weight and a 183-313% increase in panicle number compared to wild type (WT) in the first experiment. These lines also had slower leaf senescence, the so-called “stay-green” trait, indicating the KO effect of target genes in N metabolism. However, neither AAP2-KO nor CRF4-KO showed better yield or ANUE than WT. This study demonstrated the usefulness of GE technology in gene evaluation and highlighted the effects of AtAAP2 and AtCRF4 genes in the plant N cycle. El nitrógeno (N) es esencial para la síntesis de aminoácidos en la producción de arroz, pero su uso excesivo plantea una preocupación ambiental. Esta investigación tuvo como objetivo mejorar la eficiencia agronómica del uso del nitrógeno (ANUE) en el arroz mediante “knockout (KO)” de homólogos en arroz de los dos genes seleccionados de Arabidopsis thaliana: AtAAP2, una permeasa de aminoácidos involucrada en el transporte de N en los brotes, y AtCRF4, un factor de transcripción que participa en la absorción de N en las raíces. Los homólogos de estos genes en el arroz se identificaron por la similitud de la secuencia de aminoácidos y se desactivaron mediante la edición genética (GE) mediada por CRISPR/Cas9. Las líneas AAP2-KO y CRF4-KO fueron sometidas a evaluaciones agronómicas con tres dosis de N: 100% (180 kg ha-1), 50% (90 kg ha-1) y 0% (0 kg ha-1) y mostraron un aumento de 130-175% en peso de biomasa seca y un aumento de 183-313% en número de panículas en comparación con el control (WT) en el primer experimento. Estas líneas también tenían una senescencia foliar más lenta, el denominado rasgo de "permanecer verde", lo que indica el efecto de desactivación de genes del objetivo en el metabolismo del N. Sin embargo, ni AAP2-KO ni CRF4-KO mostraron mejor rendimiento o ANUE que WT. Este estudio demostró la utilidad de la tecnología de edición genética en la evaluación de genes y destacó los efectos de los genes AtAAP2 y AtCRF4 en el ciclo del N de la planta. Sociedad Colombiana de Ciencias Hortícolas-SCCH and Universidad Pedagógica y Tecnológica de Colombia-UPTC 2023-09-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Text Texto application/pdf https://revistas.uptc.edu.co/index.php/ciencias_horticolas/article/view/16120 10.17584/rcch.2023v17i3.16120 Revista Colombiana de Ciencias Hortícolas; Vol. 17 No. 3 (2023); e16120 Revista Colombiana de Ciencias Hortícolas; Vol. 17 Núm. 3 (2023); e16120 Revista Colombiana de Ciencias Hortícolas; Vol. 17 No 3 (2023); e16120 Revista Colombiana de Ciencias Hortícolas; V. 17 N. 3 (2023); e16120 Revista Colombiana de Ciencias Hortícolas; v. 17 n. 3 (2023); e16120 2422-3719 2011-2173 eng https://revistas.uptc.edu.co/index.php/ciencias_horticolas/article/view/16120/13523 Copyright (c) 2023 Revista Colombiana de Ciencias Hortícolas http://creativecommons.org/licenses/by-nc-sa/4.0 |
| spellingShingle | Amino acid permease 2 (AAP2) Cytokinin response factor 4 (CRF4) Oryza sativa L. Remote sensing Rice Nitrogen Aminoácido permeasa 2 (AAP2) Factor de respuesta de citoquinina 4 (CRF4) Oryza sativa L. Sensores remotos Arroz Nitrógeno Wakatabi, Kentaro Selvaraj, Michael Gomez Guzmán-Prada, Diego Alexander Cuásquer, Juan Bosco López-López, Karina Endo, Masaki Ishitani, Manabu Selection and evaluation of gene-edited knockout mutants of AtAAP2 and AtCRF4 homologs of rice for agronomic nitrogen use efficiency (ANUE) |
| title | Selection and evaluation of gene-edited knockout mutants of AtAAP2 and AtCRF4 homologs of rice for agronomic nitrogen use efficiency (ANUE) |
| title_alt | Selección y evaluación de mutantes knockout genéticamente editados de homólogos de AtAAP2 y AtCRF4 de arroz para la eficiencia agronómica del uso del nitrógeno (ANUE) |
| title_full | Selection and evaluation of gene-edited knockout mutants of AtAAP2 and AtCRF4 homologs of rice for agronomic nitrogen use efficiency (ANUE) |
| title_fullStr | Selection and evaluation of gene-edited knockout mutants of AtAAP2 and AtCRF4 homologs of rice for agronomic nitrogen use efficiency (ANUE) |
| title_full_unstemmed | Selection and evaluation of gene-edited knockout mutants of AtAAP2 and AtCRF4 homologs of rice for agronomic nitrogen use efficiency (ANUE) |
| title_short | Selection and evaluation of gene-edited knockout mutants of AtAAP2 and AtCRF4 homologs of rice for agronomic nitrogen use efficiency (ANUE) |
| title_sort | selection and evaluation of gene edited knockout mutants of ataap2 and atcrf4 homologs of rice for agronomic nitrogen use efficiency anue |
| topic | Amino acid permease 2 (AAP2) Cytokinin response factor 4 (CRF4) Oryza sativa L. Remote sensing Rice Nitrogen Aminoácido permeasa 2 (AAP2) Factor de respuesta de citoquinina 4 (CRF4) Oryza sativa L. Sensores remotos Arroz Nitrógeno |
| topic_facet | Amino acid permease 2 (AAP2) Cytokinin response factor 4 (CRF4) Oryza sativa L. Remote sensing Rice Nitrogen Aminoácido permeasa 2 (AAP2) Factor de respuesta de citoquinina 4 (CRF4) Oryza sativa L. Sensores remotos Arroz Nitrógeno |
| url | https://revistas.uptc.edu.co/index.php/ciencias_horticolas/article/view/16120 |
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