Abstract
Hypertension is one of the most critical risk factors accompanying cardiovascular diseases. γ-Aminobutyric acid (GABA) is a non-protein amino acid that functions as a major neurotransmitter in mammals and also as a blood-pressure lowering agent. We previously produced GABA-fortified rice lines of a popular Japonica rice cultivar ‘Koshihikari’ by genetic manipulation of GABA shunt-related genes. In the study reported here, we grew these same novel rice lines in a field trial and administered the milled rice orally to rats. The yield parameters of the transgenic rice plants were almost unchanged compared to those of untransformed cv. ‘Koshihikari’ plants, while the rice grains of the transgenic plants contained a high GABA content (3.5 g GABA/kg brown rice; 0.75–0.85 GABA g/kg milled rice) in a greenhouse trial. Oral administration of a diet containing 2.5 % GABA-fortified rice, with a daily intake for 8 weeks, had an approximately 20 mmHg anti-hypertensive effect in spontaneous hypertensive rats but not in normotensive Wistar-Kyoto rats. These results suggest that GABA-fortified rice may be applicable as a staple food to control or prevent hypertension.


Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Abe Y, Umemura S, Sugimoto K, Hirawa N, Kato Y, Yokoyama N, Yokoyama T, Iwai J, Ishii M (1995) Effect of green tea rich in gamma-aminobutyric acid on blood pressure of Dahl salt-sensitive rats. Am J Hypertens 8:74–79
Akama K, Takaiwa F (2007) C-terminal extension of rice glutamate decarboxylase (OsGAD2) functions as an autoinhibitory domain and overexpression of a truncated mutant results in the accumulation of extremely high levels of GABA in plant cells. J Exp Bot 58:2699–2707
Akama K, Akihiro T, Kitagawa M, Takaiwa F (2001) Rice (Oryza sativa) contains a novel isoform of glutamate decarboxylase that lacks an authentic calmodulin-binding domain at the C-terminus. Biochim Biophys Acta 1552:143–150
Akama K, Kanetou J, Shimosaki S, Kawakami K, Tsuchikura S, Takaiwa F (2009) Seed-specific expression of truncated OsGAD2 produces GABA-enriched rice grains that influence a decrease in blood pressure in spontaneously hypertensive rats. Transgenic Res 18:865–876
Baum G, Lev-Yadun S, Fridmann Y, Arazi T, Katsnelson H, Zik M, Fromm H (1996) Calmodulin binding to glutamate decarboxylase is required for regulation of glutamate and GABA metabolism and normal development in plants. EMBO J 15:2988–2996
Bouché N, Fromm H (2004) GABA in plants: just a metabolite? Trends Plant Sci 9:110–115
Clark SM, Di Leo R, Van Cauwenberghe OR, Mullen RT, Shelp BJ (2009) Subcellular localization and expression of multiple tomato gamma-aminobutyrate transaminases that utilize both pyruvate and glyoxylate. J Exp Bot 60:3255–3267
Costa MA, Balaszczuk AM, Domínguez A, Catanzaro O, Arranz C (1998) Effects of L-NAME and L-Arg on arterial blood pressure in normotensive and hypertensive streptozotocin diabetic rats. Acta Physiol Pharmacol Ther Latinoam 48:59–63
Curtis DR, Johnston GA (1974) Amino acid transmitters in the mammalian central nervous system. Ergeb Physiol 69:97–188
Fait A, Nesi AN, Angelovici R, Lehmann M, Pham PA, Song L, Haslam RP, Napier JA, Galili G, Fernie AR (2011) Targeted enhancement of glutamate-to-γ-aminobutyrate conversion in Arabidopsis seeds affects carbon-nitrogen balance and storage reserves in a development-dependent manner. Plant Physiol 157:1026–1042
Hayakawa K, Kimura M, Kasaha K, Matsumoto K, Sansawa H, Yamori Y (2004) Effect of a gamma-aminobutyric acid-enriched dairy product on the blood pressure of spontaneously hypertensive and normotensive Wistar-Kyoto rats. Br J Nutr 92:411–417
Hayashi A, Kimoto K (2007) Nicotianamine preferentially inhibits angiotensin I-converting enzyme. J Nutr Sci Vitaminol 53:331–336
Japan Food Research Laboratories Edit. (2007) Critical points in methods of analysis for the Japanese nutrition labeling. (In Japanese). Chuohoki Publishing, Tokyo
Kearney PM, Whelton M, Reynolds K, Muntner P, Whelton PK, He J (2005) Global burden of hypertension: analysis of worldwide data. Lancet 365:217–223
Kimura M, Hayakawa K, Sansawa H (2002) Involvement of gamma-aminobutyric acid (GABA) B receptors in the hypotensive effect of systemically administered GABA in spontaneously hypertensive rats. Jpn J Pharmacol 89:388–394
Koike S, Matsukura C, Takayama M, Asamizu E, Ezura H (2013) Suppression of γ-aminobutyric acid (GABA) transaminases induces prominent GABA accumulation, dwarfism and infertility in the tomato (Solanum lycopersicum L.). Plant Cell Physiol 54:793–807
Masuda H, Ishimaru Y, Aung MS, Kobayashi T, Kakei Y, Takahashi M, Higuchi K, Nakanishi H, Nishizawa NK (2012) Iron biofortification in rice by the introduction of multiple genes involved in iron nutrition. Sci Rep 2:543
Palanivelu R, Brass L, Edlund AF, Preuss D (2003) Pollen tube growth and guidance is regulated by POP2, an Arabidopsis gene that controls GABA levels. Cell 114:47–59
Pérez-Massot E, Banakar R, Gómez-Galera S, Zorrilla-López U, Sanahuja G, Arjó G, Miralpeix B, Vamvaka E, Farré G, Rivera SM, Dashevskaya S, Berman J, Sabalza M, Yuan D, Bai C, Bassie L, Twyman RM, Capell T, Christou P, Zhu C (2013) The contribution of transgenic plants to better health through improved nutrition: opportunities and constraints. Genes Nutr 8:29–41
Saikusa T, Horino T, Mori Y (1994) Distribution of free amino acids in the rice kernel and kernel fractions and effect of water soaking on the distribution. J Agric Food Chem 42:1122–1124
Shelp BJ, Bown AW, McLean MD (1999) Metabolism and functions of gamma-aminobutyric acid. Trends Plant Sci 4:446–452
Shimajiri Y, Ozaki K, Kainou K, Akama K (2013a) Differential subcellular localization, enzymatic properties and expression patterns of γ-aminobutyric acid transaminases (GABA-Ts) in rice (Oryza sativa). J Plant Physiol 170:196–201
Shimajiri Y, Oonishi T, Ozaki K, Kainou K, Akama K (2013b) Genetic manipulation of the γ-aminobutyric acid (GABA) shunt in rice: overexpression of truncated glutamate decarboxylase (GAD2) and knockdown of γ-aminobutyric acid transaminase (GABA-T) lead to sustained and high levels of GABA accumulation in rice kernels. Plant Biotechnol J 11:594–604
Takahashi H, Tiba M, Iino M, Takayasu T (1955) The effect of gamma-aminobutyric acid on blood pressure. Jpn J Physiol 5:334–341
Trobacher CP, Zarei A, Liu J, Clark SM, Bozzo GG, Shelp BJ (2013) Calmodulin-dependent and calmodulin-independent glutamate decarboxylases in apple fruit. BMC Plant Biol 13:144
Wakasa K, Hasegawa H, Nemoto H, Matsuda F, Miyazawa H, Tozawa Y, Morino K, Komatsu A, Yamada T, Terakawa T, Miyagawa H (2006) High-level tryptophan accumulation in seeds of transgenic rice and its limited effects on agronomic traits and seed metabolite profile. J Exp Bot 57:3069–3078
Yamakawa H, Hakata M (2010) Atlas of rice grain filling-related metabolism under high temperature: joint analysis of metabolome and transcriptome demonstrated inhibition of starch accumulation and induction of amino acid accumulation. Plant Cell Physiol 51:795–809
Yang L, Tada Y, Yamamoto MP, Zhao H, Yoshikawa M, Takaiwa F (2006) A transgenic rice seed accumulating an anti-hypertensive peptide reduces the blood pressure of spontaneously hypertensive rats. FEBS Lett 580:3315–3320
Yoshimura M, Toyoshi T, Sano A, Izumi T, Fujii T, Konishi C, Inai S, Matsukura C, Fukuda N, Ezura H, Obata A (2010) Antihypertensive effect of a gamma-aminobutyric acid rich tomato cultivar ‘DG03-9’ in spontaneously hypertensive rats. J Agric Food Chem 58:615–619
Acknowledgments
This research was supported by a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan [Genomics for Agricultural Innovation, Development of health promoting transgenic rice (GMC0005)].
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Kowaka, E., Shimajiri, Y., Kawakami, K. et al. Field trial of GABA-fortified rice plants and oral administration of milled rice in spontaneously hypertensive rats. Transgenic Res 24, 561–569 (2015). https://doi.org/10.1007/s11248-014-9859-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11248-014-9859-z


