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Evaluation of ancestral membership proportions and genotype distribution in the perception of Umami taste in Ecuadorian mestizos
Forensic Science International: Genetics Supplement Series xxx (xxxx) xxx–xxx
Contents lists available at ScienceDirect
Forensic Science International: Genetics Supplement Series journal homepage: www.elsevier.com/locate/fsigss
Evaluation of ancestral membership proportions and genotype distribution in the perception of Umami taste in Ecuadorian mestizos ⁎
C. Paz-y-Miño , J.M. García-Cárdenas, A. López-Cortés, A. Cabrera-Andrade, P. Guevara-Ramírez, D.A. González, A.K. Zambrano, P.E. Leone Centro de Investigación Genética y Genómica, Facultad de Ciencias de la Salud Eugenio Espejo, Universidad Tecnológica Equinoccial, Quito, Ecuador
A R T I C L E I N F O
A B S T R A C T
Keywords: Ecuador Umami Taste perception
Genetic variation to flavor perception contributes the population differentiation, where the T allele of SNP rs307377 of TAS1R3 gene influences the levels of appreciation of monosodium glutamate (MSG), for Umami taste. The aim of this study was to evaluate the distribution of polymorphism in the perception of Umami taste, and correlate to the ancestral proportions in the mestizo Ecuadorian population. The majority of participants presented C/C genotype. The ancestry was predominantly Native American.
1. Introduction The flavor sensation is identificated by receptor proteins that are located in the taste buds [1]. The TAS1R3 gene is found on the chromosome 1 (1p36) and encodes the receptor T1R3. The TAS1R3 gene polymorphisms (SNPs) results in different levels of taste perception [2]. Umami flavor was described in 1909 by Ikeda and is stimulated by Lglutamate (monosodium glutamate) [3]. Recent studies related the genetic factors and taste sensitivity that have characterized the populations on the perception of flavors and their choice of diet [4]. Demographically Ecuadorians have a high Native Americans ancestry. However, after Spanish colonization, Mestizo populations arose. The aim of this study was to evaluate the frequency of the polymorphisms involved in Umami taste perception in the mestizo Ecuadorian population and its ancestral composition. 2. Methodology
being N1 the lowest and N5 the highest. There were 5 types of perception using the following scale: 1–undetectable, 2–weak, 3–moderate, 4–strong and 5–very strong. 2.3. Genotyping DNA was extracted from peripheral blood using the PureLink Genomic DNA Kit (Invitrogen, Carlsbad, CA) and quantitated using NanoDrop2000 (ThermoScientific, Waltham, MA). Genotypes were amplified by PCR in a final volume of 25 μL containing 30 ng of ADN; 14.25 H2O Milli-Q; buffer 10×; MgCl2 50 mM; dNTPs each of deoxynucleotide triphosphate; 5U of Taq platinum DNA polymerase (Thermo Fisher Scientific- Invitrogen) and primers 0,2 μM. Amplicons were confirmed using electrophoresis in 2% agarose gel. Finally, they were sequenced using a BigDye Terminator Cycle Sequencing Kit v3.1 (Applied Biosystems, Austin, TX), with the Genetic Analyzer 3130 (Applied Biosystems, Austin, TX) and analyzed by using Seq-Scape Software v2.6 (Applied Biosystems).
2.1. Population 2.4. Ancestry informative markers A total of 1100 Ecuadorian individuals were randomly selected between the ages of 17 and 62. 2.2. Threshold detection evaluation To determine the phenotype, a perception test was conducted with varying concentrations of monosodium glutamate (MSG) at 1.5, 0.5, 0.1, 0.04 and 0.01%. The dilutions were classified in levels from 1 to 5,
⁎
The population was genotyped by Single Multiplex PCR using 46 autosomal Ancestry Informative Markers (AIMs). Fluorescent DNA fragments were sequenced by capillary electrophoresis in the Genetic Analyzer ABI 3130 (Applied Biosystems, Austin, TX), and were identified using the software Genemapper V 3.1 (Life Technologies, Carlsbad, CA) [5].
Corresponding author at: Avenue Mariscal Sucre, 170129 Quito, Ecuador. E-mail address: [email protected] (C. Paz-y-Miño).
http://dx.doi.org/10.1016/j.fsigss.2017.09.064 Received 25 August 2017; Accepted 13 September 2017 1875-1768/ © 2017 Elsevier B.V. All rights reserved.
Please cite this article as: Miño, C.P., Forensic Science International: Genetics Supplement Series (2017), http://dx.doi.org/10.1016/j.fsigss.2017.09.064
Forensic Science International: Genetics Supplement Series xxx (xxxx) xxx–xxx
C. Paz-y-Miño et al.
Table 1 Distribution and genotypic frequencies. Genes
Polymorphisms
Alleles
n
Genotypic
Allelic
HWE
TAS1R3
rs307377
C/C C/T T/T
1001 95 4
0,91 0,086 0.004
0,953 – 0,047
0,283
Fig. 1. Ancestral distribution of study population. The results are expressed as the averages corresponding to the 10 replicates performed in the simulation with the STRUCTURE program. Clusters 1 (African Native-American), 2 (Native-American) and 3 (European) are shown in red, green and purple respectively. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
3. Results
important to determinate dietary health risks in different ethnic populations.
3.1. Population characteristics Conflict of interest statement Of the total population, 59.10% were woman and 40.90% men. The mean age was 25 years (SD ± 1.78).
The authors have no conflict of interests.
3.2. Genotype distribution
Funding
The resulting genotyping in the Umami taste is depicted in Table 1. HWE showed balance in the polymorphisms of study.
This research did not receive any financing from public agencies, commercial, or not-for-profit sectors.
3.3. Ancestral membership proportions
Financial disclosure None.
The ancestry apportionment of all Ecuadorian participants was estimated. A greater Native American ancestry proportion (0.51) was shown, compared with the European (0.45) or the Afro-Ecuadorian groups (0.048) (Fig. 1).
Acknowledgements None.
4. Discussion References In this study, the polymorphism rs307377 (Umami), did have high prevalence of the C allele (0,953) with low influence in the perception of MSG. In comparison with a study published in 2009 by Fushan et al., determined that the rs307377 T allele polymorphism had a low level of sensitivity to MSG in Japanese population [6]. In addition, in another study they found that the presence of the T allele and the amino acid change from arginine to cysteine were a highly influential factor in reducing the perception of MSG [7]. According to Shigemura et al. (2009) the second most common genotype, in Japanese population, was the heterozygous C/T and finally the recessive homozygote T/T [8]. Besides, according to 1000 Genomes Project, phase 3 the frequency of the T allele was also close to 0 in almost all populations like African Carribbeans, Bengali from Bangladesh, Chinese and Colombians. According to Kim et al. (2006) was identified that the T allele has high frequency in population Amerindian and North American [9]. Despite fact that the population studied had greater Native American ancestry, the C allele was more prevalent than the T allele, being one of the possible causes the Spanish colonization, where the C allele prevailed in the mestizo population because of its dominance. In another study, the presence of the T allele showed a greater sensitivity to MSG in US individuals with Dutch ancestry, but the frequency of the T (0.02) allele was lower than the C (0.98) [10]. This taste perception studies are
[1] U.K. Kim, P. Breslin, D. Reed, D. Drayna, Genetics of human taste perception, J. Dent. Res. 83 (6) (2004) 448–453. [2] A.A. Bachmanov, N.P. Bosak, W.B. Floriano, M. Inoue, X. Li, C. Lin, V.O. Murovets, D.R. Reed, V.A. Zolotarev, G.K. Beauchamp, Genetics of sweet taste preferences, Flavour Fragrance J. 26 (4) (2011) 286–294. [3] B. García-Bailo, C. Toguri, K. Eny, A. El-Sohemy, Genetic variation in taste and its influence on food selection, OMICS-A J. Integr. Biol. 13 (1) (2009) 69–80. [4] C. Paz-y-Miño, G. Burgos, A. López-Cortez, C. Herrera, A. Gaviria, E. Tejera, A. Cabrera-Andrade, A study of the molecular variants associated with lactase persistence in different ecuadorian ethnic groups, Am. J. Hum. Biol. 28 (6) (2016) 774–781. [5] A. Fogel, J. Blissett, Effects of sucrose detection threshold and weight status on intake of fruit and vegetables in children, Appetite 83 (2014) 309–316. [6] A. Fushan, C. Simpson, J. Slack, A. Manichaikul, D. Drayna, Allelic Polymorphism within the TAS1R3. Promoter Is Associated with Human Taste Sensitivity to Sucrose, Current Biology 19 (15) (2009) 1288–1293. [7] B.J. Tepper, L.M. Hartfie, S.H. Schneider, Sweet taste and diet in type II diabetes, Physiol. Behav. 60 (1) (1996) 13–18. [8] N. Shigemura, S. Shirosaki, K. Sanematsu, R. Yoshida, Y. Ninomiya, Genetic and molecular basis of individual differences in human umami taste perception, PLoS One 4 (8) (2009) e6717. [9] U. Kim, S. Wooding, N. Riaz, L. Jorde, D. Drayna, Variation in the human TAS1R taste receptor genes, Chem. Senses 31 (2006) 599–611. [10] Q. Chen, S. Alarcon, A. Tharp, O. Ahmed, N. Estrella, T. Greene, J. Rucker, P. Breslin, Perceptual variation in umami taste and polymorphisms in TAS1R taste receptor genes1,2,3,4, Am. J. Clin. Nutr. 90 (3) (2009) 770S–779S.
2
Contents lists available at ScienceDirect
Forensic Science International: Genetics Supplement Series journal homepage: www.elsevier.com/locate/fsigss
Evaluation of ancestral membership proportions and genotype distribution in the perception of Umami taste in Ecuadorian mestizos ⁎
C. Paz-y-Miño , J.M. García-Cárdenas, A. López-Cortés, A. Cabrera-Andrade, P. Guevara-Ramírez, D.A. González, A.K. Zambrano, P.E. Leone Centro de Investigación Genética y Genómica, Facultad de Ciencias de la Salud Eugenio Espejo, Universidad Tecnológica Equinoccial, Quito, Ecuador
A R T I C L E I N F O
A B S T R A C T
Keywords: Ecuador Umami Taste perception
Genetic variation to flavor perception contributes the population differentiation, where the T allele of SNP rs307377 of TAS1R3 gene influences the levels of appreciation of monosodium glutamate (MSG), for Umami taste. The aim of this study was to evaluate the distribution of polymorphism in the perception of Umami taste, and correlate to the ancestral proportions in the mestizo Ecuadorian population. The majority of participants presented C/C genotype. The ancestry was predominantly Native American.
1. Introduction The flavor sensation is identificated by receptor proteins that are located in the taste buds [1]. The TAS1R3 gene is found on the chromosome 1 (1p36) and encodes the receptor T1R3. The TAS1R3 gene polymorphisms (SNPs) results in different levels of taste perception [2]. Umami flavor was described in 1909 by Ikeda and is stimulated by Lglutamate (monosodium glutamate) [3]. Recent studies related the genetic factors and taste sensitivity that have characterized the populations on the perception of flavors and their choice of diet [4]. Demographically Ecuadorians have a high Native Americans ancestry. However, after Spanish colonization, Mestizo populations arose. The aim of this study was to evaluate the frequency of the polymorphisms involved in Umami taste perception in the mestizo Ecuadorian population and its ancestral composition. 2. Methodology
being N1 the lowest and N5 the highest. There were 5 types of perception using the following scale: 1–undetectable, 2–weak, 3–moderate, 4–strong and 5–very strong. 2.3. Genotyping DNA was extracted from peripheral blood using the PureLink Genomic DNA Kit (Invitrogen, Carlsbad, CA) and quantitated using NanoDrop2000 (ThermoScientific, Waltham, MA). Genotypes were amplified by PCR in a final volume of 25 μL containing 30 ng of ADN; 14.25 H2O Milli-Q; buffer 10×; MgCl2 50 mM; dNTPs each of deoxynucleotide triphosphate; 5U of Taq platinum DNA polymerase (Thermo Fisher Scientific- Invitrogen) and primers 0,2 μM. Amplicons were confirmed using electrophoresis in 2% agarose gel. Finally, they were sequenced using a BigDye Terminator Cycle Sequencing Kit v3.1 (Applied Biosystems, Austin, TX), with the Genetic Analyzer 3130 (Applied Biosystems, Austin, TX) and analyzed by using Seq-Scape Software v2.6 (Applied Biosystems).
2.1. Population 2.4. Ancestry informative markers A total of 1100 Ecuadorian individuals were randomly selected between the ages of 17 and 62. 2.2. Threshold detection evaluation To determine the phenotype, a perception test was conducted with varying concentrations of monosodium glutamate (MSG) at 1.5, 0.5, 0.1, 0.04 and 0.01%. The dilutions were classified in levels from 1 to 5,
⁎
The population was genotyped by Single Multiplex PCR using 46 autosomal Ancestry Informative Markers (AIMs). Fluorescent DNA fragments were sequenced by capillary electrophoresis in the Genetic Analyzer ABI 3130 (Applied Biosystems, Austin, TX), and were identified using the software Genemapper V 3.1 (Life Technologies, Carlsbad, CA) [5].
Corresponding author at: Avenue Mariscal Sucre, 170129 Quito, Ecuador. E-mail address: [email protected] (C. Paz-y-Miño).
http://dx.doi.org/10.1016/j.fsigss.2017.09.064 Received 25 August 2017; Accepted 13 September 2017 1875-1768/ © 2017 Elsevier B.V. All rights reserved.
Please cite this article as: Miño, C.P., Forensic Science International: Genetics Supplement Series (2017), http://dx.doi.org/10.1016/j.fsigss.2017.09.064
Forensic Science International: Genetics Supplement Series xxx (xxxx) xxx–xxx
C. Paz-y-Miño et al.
Table 1 Distribution and genotypic frequencies. Genes
Polymorphisms
Alleles
n
Genotypic
Allelic
HWE
TAS1R3
rs307377
C/C C/T T/T
1001 95 4
0,91 0,086 0.004
0,953 – 0,047
0,283
Fig. 1. Ancestral distribution of study population. The results are expressed as the averages corresponding to the 10 replicates performed in the simulation with the STRUCTURE program. Clusters 1 (African Native-American), 2 (Native-American) and 3 (European) are shown in red, green and purple respectively. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
3. Results
important to determinate dietary health risks in different ethnic populations.
3.1. Population characteristics Conflict of interest statement Of the total population, 59.10% were woman and 40.90% men. The mean age was 25 years (SD ± 1.78).
The authors have no conflict of interests.
3.2. Genotype distribution
Funding
The resulting genotyping in the Umami taste is depicted in Table 1. HWE showed balance in the polymorphisms of study.
This research did not receive any financing from public agencies, commercial, or not-for-profit sectors.
3.3. Ancestral membership proportions
Financial disclosure None.
The ancestry apportionment of all Ecuadorian participants was estimated. A greater Native American ancestry proportion (0.51) was shown, compared with the European (0.45) or the Afro-Ecuadorian groups (0.048) (Fig. 1).
Acknowledgements None.
4. Discussion References In this study, the polymorphism rs307377 (Umami), did have high prevalence of the C allele (0,953) with low influence in the perception of MSG. In comparison with a study published in 2009 by Fushan et al., determined that the rs307377 T allele polymorphism had a low level of sensitivity to MSG in Japanese population [6]. In addition, in another study they found that the presence of the T allele and the amino acid change from arginine to cysteine were a highly influential factor in reducing the perception of MSG [7]. According to Shigemura et al. (2009) the second most common genotype, in Japanese population, was the heterozygous C/T and finally the recessive homozygote T/T [8]. Besides, according to 1000 Genomes Project, phase 3 the frequency of the T allele was also close to 0 in almost all populations like African Carribbeans, Bengali from Bangladesh, Chinese and Colombians. According to Kim et al. (2006) was identified that the T allele has high frequency in population Amerindian and North American [9]. Despite fact that the population studied had greater Native American ancestry, the C allele was more prevalent than the T allele, being one of the possible causes the Spanish colonization, where the C allele prevailed in the mestizo population because of its dominance. In another study, the presence of the T allele showed a greater sensitivity to MSG in US individuals with Dutch ancestry, but the frequency of the T (0.02) allele was lower than the C (0.98) [10]. This taste perception studies are
[1] U.K. Kim, P. Breslin, D. Reed, D. Drayna, Genetics of human taste perception, J. Dent. Res. 83 (6) (2004) 448–453. [2] A.A. Bachmanov, N.P. Bosak, W.B. Floriano, M. Inoue, X. Li, C. Lin, V.O. Murovets, D.R. Reed, V.A. Zolotarev, G.K. Beauchamp, Genetics of sweet taste preferences, Flavour Fragrance J. 26 (4) (2011) 286–294. [3] B. García-Bailo, C. Toguri, K. Eny, A. El-Sohemy, Genetic variation in taste and its influence on food selection, OMICS-A J. Integr. Biol. 13 (1) (2009) 69–80. [4] C. Paz-y-Miño, G. Burgos, A. López-Cortez, C. Herrera, A. Gaviria, E. Tejera, A. Cabrera-Andrade, A study of the molecular variants associated with lactase persistence in different ecuadorian ethnic groups, Am. J. Hum. Biol. 28 (6) (2016) 774–781. [5] A. Fogel, J. Blissett, Effects of sucrose detection threshold and weight status on intake of fruit and vegetables in children, Appetite 83 (2014) 309–316. [6] A. Fushan, C. Simpson, J. Slack, A. Manichaikul, D. Drayna, Allelic Polymorphism within the TAS1R3. Promoter Is Associated with Human Taste Sensitivity to Sucrose, Current Biology 19 (15) (2009) 1288–1293. [7] B.J. Tepper, L.M. Hartfie, S.H. Schneider, Sweet taste and diet in type II diabetes, Physiol. Behav. 60 (1) (1996) 13–18. [8] N. Shigemura, S. Shirosaki, K. Sanematsu, R. Yoshida, Y. Ninomiya, Genetic and molecular basis of individual differences in human umami taste perception, PLoS One 4 (8) (2009) e6717. [9] U. Kim, S. Wooding, N. Riaz, L. Jorde, D. Drayna, Variation in the human TAS1R taste receptor genes, Chem. Senses 31 (2006) 599–611. [10] Q. Chen, S. Alarcon, A. Tharp, O. Ahmed, N. Estrella, T. Greene, J. Rucker, P. Breslin, Perceptual variation in umami taste and polymorphisms in TAS1R taste receptor genes1,2,3,4, Am. J. Clin. Nutr. 90 (3) (2009) 770S–779S.
2