Efficient increase of ɣ-aminobutyric acid (GABA) content in tomato fruits by targeted mutagenesis
Open accessAbstract
γ-Aminobutyric acid (GABA) is a non-proteinogenic amino acid that has hypotensive effects. Tomato ( Solanum lycopersicum L.) is among the most widely cultivated and consumed vegetables in the world and contains higher levels of GABA than other major crops. Increasing these levels can further enhance the blood pressure-lowering function of tomato fruit. Glutamate decarboxylase (GAD) is a key enzyme in GABA biosynthesis; it has a C-terminal autoinhibitory domain that regulates enzymatic function, and deleting this domain increases GAD activity. The tomato genome has five GAD genes ( SlGAD1 – 5 ), of which two ( SlGAD2 and SlGAD3 ) are expressed during tomato fruit development. To increase GABA content in tomato, we deleted the autoinhibitory domain of SlGAD2 and SlGAD3 using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas)9 technology. Introducing a stop codon immediately before the autoinhibitory domain increased GABA accumulation by 7 to 15 fold while having variable effects on plant and fruit size and yield. This is the first study describing the application of the CRISPR/Cas9 system to increase GABA content in tomato fruits. Our findings provide a basis for the improvement of other types of crop by CRISPR/Cas9-based genetic modification.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Mutagenesis is more acceptable than transgenesis in breeding, yet in some studies its efficiency is reported to be low.
“Mutagenesis is more acceptable than transgenesis in breeding, yet in some studies its efficiency is reported to be low (Nonaka et al., 2017).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Nonaka et al., 2017
Introduction of CRISPR as a gene-editing platform has greatly improved the efficiency of breeding.
“The introduction of CRISPR as a gene-editing platform has greatly improved the efficiency of breeding (Nonaka et al., 2017) and the functional characterization of genes (Zhang et al., 2021;Karunarathne et al., 2022).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Nonaka et al., 2017
Few developments using CRISPR/Cas9 have reached commercialization.
“There have been numerous proof-of-concept experiments conducted using the CRISPR/Cas9 gene-editing platform (Lawrenson et al., 2015), but few developments have reached commercialization (Nonaka et al., 2017).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Nonaka et al., 2017
Introduction of a stop codon to the Solanum lycopersicum L. glutamate decarboxylase SlGAD2 and SlGAD3 genes through CRISPR/Cas9 technology increased the GABA content in the tomatoes by 7-fold to 15-fold.
“Introduction of a stop codon to the Solanum lycopersicum L. glutamate decarboxylase SlGAD2 and SlGAD3 genes through CRISPR/ Cas9 technology increased the GABA content in the tomatoes by 7-fold to 15-fold (Nonaka et al., 2017).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Nonaka et al., 2017
Sanger sequencing is widely used and useful for low-volume high-quality DNA sequencing.
“Sanger sequencing is widely used and useful for low-volume high-quality DNA sequencing (Nonaka et al., 2017;Doll et al., 2019).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Nonaka et al., 2017
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012) · Science · doi:10.1126/science.1225829 · Cited by 17,116 · see citing works
Multiplex Genome Engineering Using CRISPR/Cas Systems
Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A., et al. (2013) · Science · doi:10.1126/science.1231143 · Cited by 15,339 · see citing works
Zhou, B., Bentham, J., Di Cesare, M., Bixby, H., Danaei, G., Cowan, M. J., Paciorek, C. J., Singh, G., Hajifathalian, K., Bennett, J. E., et al. (2017) · The Lancet · doi:10.1016/s0140-6736(16)31919-5 · Cited by 2,579 · see citing works
Jiang, W., Zhou, H., Bi, H., Fromm, M., Yang, B., & Weeks, D. P. (2013) · Nucleic Acids Research · doi:10.1093/nar/gkt780 · Cited by 1,250 · see citing works
GABA in plants: just a metabolite?
Bouché, N., & Fromm, H. (2004) · Trends in Plant Science · doi:10.1016/j.tplants.2004.01.006 · Cited by 1,170 · see citing works
Highly Efficient Targeted Mutagenesis of Drosophila with the CRISPR/Cas9 System
Bassett, A. R., Tibbit, C., Ponting, C. P., & Liu, J. (2013) · Cell Reports · doi:10.1016/j.celrep.2013.06.020 · Cited by 849 · see citing works
Genome editing with RNA-guided Cas9 nuclease in Zebrafish embryos
Chang, N., Sun, C., Gao, L., Zhu, D., Xu, X., Zhu, X., Xiong, J., & Xi, J. (2013) · Cell Research · doi:10.1038/cr.2013.45 · Cited by 819 · see citing works
Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y., Yang, L., Zhang, H., Xu, N., et al. (2014) · Plant Biotechnology Journal · doi:10.1111/pbi.12200 · Cited by 799 · see citing works
Brooks, C., Nekrasov, V., Lippman, Z. B., & Eck, J. M. V. (2014) · PLANT PHYSIOLOGY · doi:10.1104/pp.114.247577 · Cited by 759 · see citing works
Efficient octopine Ti plasmid-derived vectors for Agrobacterium -mediated gene transfer to plants
Deblaere, R., Bytebier, B., Greve, H. D., et al. (1985) · Nucleic Acids Research · doi:10.1093/nar/13.13.4777 · Cited by 716 · see citing works
Fauser, F., Schiml, S., & Puchta, H. (2014) · The Plant Journal · doi:10.1111/tpj.12554 · Cited by 704 · see citing works
Inoue, K., Shirai, T., Ochiai, H., Kasao, M., Hayakawa, K., Kimura, M., & Sansawa, H. (2003) · European Journal of Clinical Nutrition · doi:10.1038/sj.ejcn.1601555 · Cited by 546 · see citing works
Sun, H., Uchii, S., Watanabe, S., & Ezura, H. (2005) · Plant and Cell Physiology · doi:10.1093/pcp/pci251 · Cited by 476 · see citing works
Self-citationThe Metabolism and Functions of [gamma]-Aminobutyric Acid
Bown, A. W., & Shelp, B. J. (1997) · PLANT PHYSIOLOGY · doi:10.1104/pp.115.1.1 · Cited by 398 · see citing works
Pan, C., Ye, L., Li, Q., Liu, X., He, Y., Wang, J., Chen, L., & Lu, G. (2016) · Scientific Reports · doi:10.1038/srep24765 · Cited by 381 · see citing works
CRISPR/Cas9-mediated mutagenesis of the RIN locus that regulates tomato fruit ripening
Ito, Y., Nishizawa‐Yokoi, A., Endo, M., Mikami, M., & Toki, S. (2015) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2015.09.117 · Cited by 328 · see citing works
Baum, G., Lev‐Yadun, S., Fridmann, Y., et al. (1996) · The EMBO Journal · doi:10.1002/j.1460-2075.1996.tb00662.x · Cited by 306 · see citing works
Comparison of CRISPR/Cas9 expression constructs for efficient targeted mutagenesis in rice
Mikami, M., Toki, S., & Endo, M. (2015) · Plant Molecular Biology · doi:10.1007/s11103-015-0342-x · Cited by 290 · see citing works
TOMATOMA: A Novel Tomato Mutant Database Distributing Micro-Tom Mutant Collections
Saito, T., Ariizumi, T., Okabe, Y., et al. (2011) · Plant and Cell Physiology · doi:10.1093/pcp/pcr004 · Cited by 256 · see citing works
Baum, G., Chen, Y., Arazi, T., et al. (1993) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)36560-3 · Cited by 232 · see citing works
Biochemical Mechanism on GABA Accumulation During Fruit Development in Tomato
Akihiro, T., Koike, S., Tani, R., Tominaga, T., Watanabe, S., Iijima, Y., Aoki, K., Shibata, D., Ashihara, H., Matsukura, C., et al. (2008) · Plant and Cell Physiology · doi:10.1093/pcp/pcn113 · Cited by 209 · see citing works
Self-citationPark, K.-B., & Oh, S. (2006) · Bioresource Technology · doi:10.1016/j.biortech.2006.06.006 · Cited by 204 · see citing works
Okabe, Y., Asamizu, E., Saito, T., et al. (2011) · Plant and Cell Physiology · doi:10.1093/pcp/pcr134 · Cited by 200 · see citing works
Elliott, K. A. C., & Jasper, H. H. (1959) · Physiological Reviews · doi:10.1152/physrev.1959.39.2.383 · Cited by 189 · see citing works
Saikusa, T., Horino, T., & Mori, Y. (1994) · Journal of Agricultural and Food Chemistry · doi:10.1021/jf00041a015 · Cited by 183 · see citing works
Renault, H., Amrani, A. E., Palanivelu, R., et al. (2011) · Plant and Cell Physiology · doi:10.1093/pcp/pcr041 · Cited by 159 · see citing works
Fait, A., Nunes‐Nesi, A., Angelovici, R., Lehmann, M., Pham, P. A., Song, L., Haslam, R. P., Napier, J. A., Galili, G., & Fernie, A. R. (2011) · PLANT PHYSIOLOGY · doi:10.1104/pp.111.179986 · Cited by 146 · see citing works
Elliott, K. A. C., & Hobbiger, F. (1959) · The Journal of Physiology · doi:10.1113/jphysiol.1959.sp006178 · Cited by 142 · see citing works
Aoki, H., Ichiyo, U. D. A., Tagami, K., Furuya, Y., Endo, Y., & Fujimoto, K. (2003) · Bioscience Biotechnology and Biochemistry · doi:10.1271/bbb.67.1018 · Cited by 141 · see citing works
Arazi, T., Baum, G., Snedden, W. A., et al. (1995) · PLANT PHYSIOLOGY · doi:10.1104/pp.108.2.551 · Cited by 134 · see citing works
Omori, M., Yano, T., Okamoto, J., Tsushida, T., MURAI, T., & Higuchi, M. (1987) · Nippon Nōgeikagaku Kaishi · doi:10.1271/nogeikagaku1924.61.1449 · Cited by 121 · see citing works
Yap, K. L., Yuan, T., Mal, T. K., Vogel, H. J., & Ikura, M. (2003) · Journal of Molecular Biology · doi:10.1016/s0022-2836(03)00271-7 · Cited by 112 · see citing works
Akama, K., & Takaiwa, F. (2007) · Journal of Experimental Botany · doi:10.1093/jxb/erm120 · Cited by 110 · see citing works
Gut, H., Dominici, P., Pilati, S., Astegno, A., Petoukhov, M. V., Svergun, D. I., Grütter, M. G., & Capitani, G. (2009) · Journal of Molecular Biology · doi:10.1016/j.jmb.2009.06.080 · Cited by 98 · see citing works
Conversion of Glutamic Acid to γ -Aminobutyric Acid in Tea Leaves under Anaerobic Conditions
Tsushida, T., & MURAI, T. (1987) · Agricultural and Biological Chemistry · doi:10.1080/00021369.1987.10868498 · Cited by 94 · see citing works
THE EFFECT OF γ-AMINOBUIC ACID ON BLOOD PRESSURE
Takahashi, H., TIBA, M., IINO, M., et al. (1955) · The Japanese Journal of Physiology · doi:10.2170/jjphysiol.5.334 · Cited by 88 · see citing works
Takayama, M., Koike, S., Kusano, M., et al. (2015) · Plant and Cell Physiology · doi:10.1093/pcp/pcv075 · Cited by 72 · see citing works
Self-citation[104a] Enzymes of γ-aminobutyrate metabolism (bacterial)
Jakoby, W. B. (1962) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/s0076-6879(62)05311-2 · Cited by 71 · see citing works
Calmodulin-dependent and calmodulin-independent glutamate decarboxylases in apple fruit
Trobacher, C. P., Zarei, A., Liu, J., et al. (2013) · BMC Plant Biology · doi:10.1186/1471-2229-13-144 · Cited by 70 · see citing works
Koike, S., Matsukura, C., Takayama, M., et al. (2013) · Plant and Cell Physiology · doi:10.1093/pcp/pct035 · Cited by 68 · see citing works
Self-citationYamakoshi, J., Fukuda, S., SATOH, T., 辻亮平, SAITO, M., Obata, A., Matsuyama, A., Kikuchi, M., & Kawasaki, T. (2007) · Bioscience Biotechnology and Biochemistry · doi:10.1271/bbb.60424 · Cited by 62 · see citing works
NMR study of low subcellular pH during the development of cherry tomato fruit
Rolin, D., Baldet, P., Just, D., et al. (2000) · Australian Journal of Plant Physiology · doi:10.1071/pp99051 · Cited by 58 · see citing works
Yuan, T., & Vogel, H. J. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.46.30328 · Cited by 58 · see citing works
Takayama, M., Matsukura, C., Ariizumi, T., et al. (2016) · Plant Cell Reports · doi:10.1007/s00299-016-2061-4 · Cited by 54 · see citing works
Self-citationThe Transcription Factor E4F1 Coordinates CHK1-Dependent Checkpoint and Mitochondrial Functions
Rodier, G., Kirsh, O., Baraibar, M. A., Houlès, T., Lacroix, M., Delpech, H., Hatchi, E., Arnould, S., Séverac, D., Dubois, E., et al. (2015) · Cell Reports · doi:10.1016/j.celrep.2015.03.024 · Cited by 52 · see citing works
EFFECT OF γ-AMINOBUTYRIC ACID (GABA) ON NORMOTENSIVE OR HYPERTENSIVE RATS AND MEN
Takahashi, H., Sumi, M., & KOSHINO, F. (1961) · The Japanese Journal of Physiology · doi:10.2170/jjphysiol.11.89 · Cited by 44 · see citing works
Zik, M., Fridmann‐Sirkis, Y., & Fromm, H. (2006) · Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics · doi:10.1016/j.bbapap.2006.02.007 · Cited by 19 · see citing works
KAZAMI, D., Ogura, N., Fukuchi, T., et al. (2002) · NIPPON SHOKUHIN KOGYO GAKKAISHI · doi:10.3136/nskkk.49.409 · Cited by 13 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
The CRISPR/Cas9 system and its applications in crop genome editing
Bao, A., Burritt, D. J., Chen, H., Zhou, X., Cao, D., & Tran, L.-S. P. (2019) · Critical Reviews in Biotechnology · doi:10.1080/07388551.2018.1554621 · Cited by 185 · see citing works
Genetic Variation and Unintended Risk in the Context of Old and New Breeding Techniques
Singer, S. D., Laurie, J. D., Bilichak, A., Kumar, S., & Singh, J. (2021) · Critical Reviews in Plant Sciences · doi:10.1080/07352689.2021.1883826 · Cited by 77 · see citing works
Concerns regarding ‘off-target’activity of genome editing endonucleases
Kadam, U. S., Shelake, R. M., Chavhan, R. L., & Suprasanna, P. (2018) · Plant Physiology and Biochemistry · doi:10.1016/j.plaphy.2018.03.027 · Cited by 47 · see citing works
Karunarathne, S., Walker, E., Sharma, D., Li, C., & Han, Y. (2023) · Journal of Zhejiang University-SCIENCE B · doi:10.1631/jzus.b2200552 · Cited by 18 · see citing works
“There have been numerous proof-of-concept experi‐ ments conducted using the CRISPR/Cas9 gene-editing platform (Lawrenson et al., 2015), but few devel‐ opments have reached commercialization (Nonaka et al., 2017).”
Influential
- Authors
- Nonaka, S., Arai, C., Takayama, M., Matsukura, C., & Ezura, H.
- Affiliations
- University of Tsukuba (JP)
- Corresponding author
- Ezura, Hiroshi
- Venue
- Scientific Reports
- Published
- 2017-08-01
- Volume
- 7
- Issue
- 1
- Pages
- 7057-7057
- ISSN
- 2045-2322 (ISSN-L)
- PMC
- PMC5539196
- OpenAlex
- W2738219005
- Cited by
- 357
- References
- 48
- Citations by year
- 2026: 40 · 2025: 54 · 2024: 63 · 2023: 53 · 2022: 46 · 2021: 36 · 2020: 32 · 2019: 16 · 2018: 15 · 2017: 2
- FWCI
- 17.49
- Type
- journal article
- Language
- en
- Update notices
- correction (2019-12-19)
- Open access
- gold
- doi.org
- Locations
- Scientific Reports (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Institutional Repositories DataBase (IRDB) (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Terrestrial Environment Research Center (University of Tsukuba) (submittedVersion, cc-by)
- Funders
- Cabinet Office, Government of Japan
- University of Tsukuba
- Japan Society for the Promotion of Science (15K07253)
- Bio-oriented Technology Research Advancement Institution
- Topics
- Life Sciences › Agricultural and Biological Sciences › Plant Science › GABA and Rice Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Genetically Modified Organisms Research
- Keywords
- GABA, tomato fruit, glutamic acid decarboxylase, CRISPR/Cas9 mutagenesis, GABA synthesis, fruit development, crop improvement
- MeSH terms
- Gene Editing, CRISPR-Associated Protein 9, Fruit, gamma-Aminobutyric Acid, Glutamate Decarboxylase, Mutagenesis, Sequence Deletion, Solanum lycopersicum, Metabolic Networks and Pathways, Metabolic Engineering, Clustered Regularly Interspaced Short Palindromic Repeats
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
