The grape variety Sauvignon Blanc (SB) is the flagship of New Zealand’s wine industry and accounted for over 75 % of the value of total wine exports in 2008. Two volatile thiols, 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl-acetate (3MHA), reminiscent of grapefruit and passion fruit respectively, are critical for the main varietal characters in New Zealand SB. These aromatic thiols are not present in the grape juice, but are synthesized and released by the yeast during alcoholic fermentation from non-aromatic precursors. The aim of this work was to elucidate the underlying genetics of volatile thiol synthesis in yeast (Saccharomyces cerevisiae) during alcoholic fermentation of grape juice. A gene-deletion strategy was chosen for the investigation of putative genes influencing 3MH and 3MHA release. The first part of this thesis optimized fermentation conditions in grape-juice-based media, which enabled auxotrophic laboratory strains, derived from S288C, to ferment grape juice to completion with high efficiency. Key steps to achieving this goal were the supplementation of the grape juice with higher than recommended amounts of amino acids, which increased the fermentation rate of auxotrophic yeast strains. Lysine auxotrophic strains especially benefited from this measure. In combination with the dilution of SB grape juice by 25 % with synthetic grape juice without sugars, the auxotrophic laboratory yeast BY4743 was able to metabolize all sugars in the grape juice-based media in a time frame similar to that of a commercial wine yeast. The key properties of the resulting wine were comparable to wine made with a commercial wine yeast under the same conditions. In the second part, these newly developed fermentation conditions were employed to screen 69 single-gene deletion strains in the laboratory yeast background BY4743. The list of the 69 candidate genes was compiled by combining existing knowledge about thiol production in yeast with the mining of several biological databases. Screening of the single-gene deletions revealed 17 genes which caused biologically relevant increases or decreases in volatile thiol production, but none abolished it. The majority of the 17 genes were related to the sulfur and nitrogen metabolism in yeast. A subset of these thiol-influencing genes were also deleted in a wine yeast, and were overexpressed in both wine yeast and laboratory yeast, to gain more insight in their regulatory effects. The findings confirmed that sulfur and nitrogen metabolism in yeast were important in regulating 3MH and 3MHA synthesis. Different sulfur and nitrogen sources were added to the grape must prior to fermentation and their effect on thiol release was studied. It was found that nitrogen sources urea and DAP, as well as, the sulfur compound S-ethyl-L-cysteine (SEC) increased 3MH and 3MHA concentrations in the resulting wines. The addition of cysteine to grape juice fermented with wine yeast deleted in genes CYS3 and CYS4 more than doubled total thiol production. Mapping approaches to investigate thiol production in yeast were employed in the final part of this thesis. Genetically mapped F2 progeny of a cross between a low thiol-producing yeast strain and a high-thiol producer were screened for their thiol phenotype. The 3MH and 3MHA phenotypes across 48 screened F2 progeny resembled normal distributions, indicating a quantitative trait. Subsequent mapping identified a locus on chromosome 14 with a small effect on the 3MHA phenotype, but no obvious candidate genes were evident in the region. Another approach to investigate the evolution of volatile thiols in yeast included the use of SEC, a thiol compound resembling the cysteinylated precursor of 3MH, as a sole nitrogen source in a yeast growth assay. It was found that most wine yeast, European yeast isolates and laboratory yeasts could utilize SEC as a nitrogen source, whereas various other S. cerevisiae isolates could not. Crosses between three pairs of Sec- and Sec+ yeast strains strongly indicated that this trait was monogenically inherited. However, no direct correlation between the SEC phenotype and volatile release could be observed. Genetic mapping experiments in one SEC-segregating yeast population linked this SEC phenotype to the leu2-D0 deletion in a cross between a Leu+ and Leu- yeast strain. It was shown that leucine auxotrophy most likely caused the Sec- phenotype. In a second F2 population of a cross between prototrophic Sec+ and Sec- strains, strong linkage was established to a region on chromosome 6 containing two candidate genes, DUG1 and IRC7. DUG1 was proved not to be the cause of the SEC phenotype, whereas IRC7 remains a strong candidate gene.

The grape variety Sauvignon Blanc (SB) is the flagship of New Zealand’s wine industry and accounted for over 75 % of the value of total wine exports in 2008. Two volatile thiols, 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl-acetate (3MHA), reminiscent of grapefruit and passion fruit respectively, are critical for the main varietal characters in New Zealand SB. These aromatic thiols are not present in the grape juice, but are synthesized and released by the yeast during alcoholic fermentation from non-aromatic precursors. The aim of this work was to elucidate the underlying genetics of volatile thiol synthesis in yeast (Saccharomyces cerevisiae) during alcoholic fermentation of grape juice. A gene-deletion strategy was chosen for the investigation of putative genes influencing 3MH and 3MHA release. The first part of this thesis optimized fermentation conditions in grape-juice-based media, which enabled auxotrophic laboratory strains, derived from S288C, to ferment grape juice to completion with high efficiency. Key steps to achieving this goal were the supplementation of the grape juice with higher than recommended amounts of amino acids, which increased the fermentation rate of auxotrophic yeast strains. Lysine auxotrophic strains especially benefited from this measure. In combination with the dilution of SB grape juice by 25 % with synthetic grape juice without sugars, the auxotrophic laboratory yeast BY4743 was able to metabolize all sugars in the grape juice-based media in a time frame similar to that of a commercial wine yeast. The key properties of the resulting wine were comparable to wine made with a commercial wine yeast under the same conditions. In the second part, these newly developed fermentation conditions were employed to screen 69 single-gene deletion strains in the laboratory yeast background BY4743. The list of the 69 candidate genes was compiled by combining existing knowledge about thiol production in yeast with the mining of several biological databases. Screening of the single-gene deletions revealed 17 genes which caused biologically relevant increases or decreases in volatile thiol production, but none abolished it. The majority of the 17 genes were related to the sulfur and nitrogen metabolism in yeast. A subset of these thiol-influencing genes were also deleted in a wine yeast, and were overexpressed in both wine yeast and laboratory yeast, to gain more insight in their regulatory effects. The findings confirmed that sulfur and nitrogen metabolism in yeast were important in regulating 3MH and 3MHA synthesis. Different sulfur and nitrogen sources were added to the grape must prior to fermentation and their effect on thiol release was studied. It was found that nitrogen sources urea and DAP, as well as, the sulfur compound S-ethyl-L-cysteine (SEC) increased 3MH and 3MHA concentrations in the resulting wines. The addition of cysteine to grape juice fermented with wine yeast deleted in genes CYS3 and CYS4 more than doubled total thiol production. Mapping approaches to investigate thiol production in yeast were employed in the final part of this thesis. Genetically mapped F2 progeny of a cross between a low thiol-producing yeast strain and a high-thiol producer were screened for their thiol phenotype. The 3MH and 3MHA phenotypes across 48 screened F2 progeny resembled normal distributions, indicating a quantitative trait. Subsequent mapping identified a locus on chromosome 14 with a small effect on the 3MHA phenotype, but no obvious candidate genes were evident in the region. Another approach to investigate the evolution of volatile thiols in yeast included the use of SEC, a thiol compound resembling the cysteinylated precursor of 3MH, as a sole nitrogen source in a yeast growth assay. It was found that most wine yeast, European yeast isolates and laboratory yeasts could utilize SEC as a nitrogen source, whereas various other S. cerevisiae isolates could not. Crosses between three pairs of Sec- and Sec+ yeast strains strongly indicated that this trait was monogenically inherited. However, no direct correlation between the SEC phenotype and volatile release could be observed. Genetic mapping experiments in one SEC-segregating yeast population linked this SEC phenotype to the leu2-D0 deletion in a cross between a Leu+ and Leu- yeast strain. It was shown that leucine auxotrophy most likely caused the Sec- phenotype. In a second F2 population of a cross between prototrophic Sec+ and Sec- strains, strong linkage was established to a region on chromosome 6 containing two candidate genes, DUG1 and IRC7. DUG1 was proved not to be the cause of the SEC phenotype, whereas IRC7 remains a strong candidate gene.

ABSTRACT A core tool of sensory science is the use of trained descriptive panels. This research describes an investigation into the role of motivation in the performance of trained panels and the use of a trained panel to develop a better understanding of the perception of Sauvignon blanc wines. Substantial investment in time and money is directed towards ensuring trained panels perform optimally. Having selected a panel, the panel leader needs to ensure that panellists provide accurate, reliable data. Panellist motivation is also an important factor to consider. While performance psychology, education and sport science fields have researched motivation extensively, knowledge about panellist motivation within sensory science is limited. However, findings from existing research in these other areas - which suggest an important role for autonomy, competence and relatedness - can be applied to sensory panels in order to increase intrinsic motivation. The initial part of the research investigated the fundamental factors that affect and influence panellists’ motivation and participation. A survey (n=74) revealed that extra income and a general interest in food were the key drivers in inspiring people to become panellists, whilst enjoyment in being a panellist, interest in food, and extra income were key drivers for people to remain panellists. In a second survey, the intrinsic motivation of seven trained panels from four countries (n=108) was assessed. External panels were found to be more intrinsically motivated than internal panels. Experienced panellists had an increased perception of competence, which is a key factor for people to be intrinsically motivated. Understanding motivational frameworks currently used in other research fields and integrating them into existing panel training protocols may enhance and sustain panellists’ intrinsic motivation. A trained panel (n=14) was then used in the second part of the thesis to identify key flavours in Sauvignon blanc wines from Australia, France, New Zealand, Spain, South Africa and USA. Sixteen characteristics were identified and measured, including sweet sweaty passionfruit, capsicum, passionfruit skin/stalk, boxwood/cat’s urine, grassy, mineral/flinty, citrus, bourbon, apple lolly/candy, tropical, mint, fresh asparagus, canned asparagus, stonefruit, apple and snowpea. Principal component analysis was used to describe differences between regions and countries. Sauvignon blanc wines from Marlborough, New Zealand (NZ), were described by tropical and sweet sweaty passionfruit characteristics, while French and South African Sauvignon blanc wines were described as having flinty/mineral and bourbon-like flavors. Chemical analyses of these wines also showed that Marlborough, NZ wines had more methoxypyrazine and thiol compounds. A consumer study (n=109) showed that New Zealanders significantly prefer New Zealand style Sauvignon blanc. The final part of this research focused on using trained panellists to explore the interactions between volatile and non-volatile wine compounds and their effects on the aroma profile of New Zealand Sauvignon blanc wine. Four volatile aroma compounds that are important in New Zealand Sauvignon blanc wine were studied (isobutyl methoxypyrazine [MIBP], 3-mercaptohexanol [3MH], 3-mercaptohexanol acetate [3MHA], and ethyl decanoate). Each of these four aroma compounds were assessed in combination with three non-volatile polyphenolic compounds commonly found in Sauvignon blanc wine: catechin, caffeic acid and quercetin. Results showed each polyphenol had a unique effect when blended with a specific aroma compound, either suppressing, accentuating, or showing little effect on the perception of the aroma compounds. The perception of MIBP, 3MH, and ethyl decanoate were largely suppressed by the added polyphenols, with a few exceptions. The perception of 3MH was accentuated with the addition of caffeic acid, and the perception of 3MHA was accentuated with the addition of catechin. The interactive effects of aroma compounds with polyphenols likely reflect non-covalent associations in the wine solution that reduce the volatility of the aroma compounds. With an understanding of the interactive effects of volatile and non-volatile compounds in wine, winemakers might optimize the impact of selected volatile compounds by managing polyphenol levels, supporting their efforts to attain desirable wine aroma profiles.

ABSTRACT A core tool of sensory science is the use of trained descriptive panels. This research describes an investigation into the role of motivation in the performance of trained panels and the use of a trained panel to develop a better understanding of the perception of Sauvignon blanc wines. Substantial investment in time and money is directed towards ensuring trained panels perform optimally. Having selected a panel, the panel leader needs to ensure that panellists provide accurate, reliable data. Panellist motivation is also an important factor to consider. While performance psychology, education and sport science fields have researched motivation extensively, knowledge about panellist motivation within sensory science is limited. However, findings from existing research in these other areas - which suggest an important role for autonomy, competence and relatedness - can be applied to sensory panels in order to increase intrinsic motivation. The initial part of the research investigated the fundamental factors that affect and influence panellists’ motivation and participation. A survey (n=74) revealed that extra income and a general interest in food were the key drivers in inspiring people to become panellists, whilst enjoyment in being a panellist, interest in food, and extra income were key drivers for people to remain panellists. In a second survey, the intrinsic motivation of seven trained panels from four countries (n=108) was assessed. External panels were found to be more intrinsically motivated than internal panels. Experienced panellists had an increased perception of competence, which is a key factor for people to be intrinsically motivated. Understanding motivational frameworks currently used in other research fields and integrating them into existing panel training protocols may enhance and sustain panellists’ intrinsic motivation. A trained panel (n=14) was then used in the second part of the thesis to identify key flavours in Sauvignon blanc wines from Australia, France, New Zealand, Spain, South Africa and USA. Sixteen characteristics were identified and measured, including sweet sweaty passionfruit, capsicum, passionfruit skin/stalk, boxwood/cat’s urine, grassy, mineral/flinty, citrus, bourbon, apple lolly/candy, tropical, mint, fresh asparagus, canned asparagus, stonefruit, apple and snowpea. Principal component analysis was used to describe differences between regions and countries. Sauvignon blanc wines from Marlborough, New Zealand (NZ), were described by tropical and sweet sweaty passionfruit characteristics, while French and South African Sauvignon blanc wines were described as having flinty/mineral and bourbon-like flavors. Chemical analyses of these wines also showed that Marlborough, NZ wines had more methoxypyrazine and thiol compounds. A consumer study (n=109) showed that New Zealanders significantly prefer New Zealand style Sauvignon blanc. The final part of this research focused on using trained panellists to explore the interactions between volatile and non-volatile wine compounds and their effects on the aroma profile of New Zealand Sauvignon blanc wine. Four volatile aroma compounds that are important in New Zealand Sauvignon blanc wine were studied (isobutyl methoxypyrazine [MIBP], 3-mercaptohexanol [3MH], 3-mercaptohexanol acetate [3MHA], and ethyl decanoate). Each of these four aroma compounds were assessed in combination with three non-volatile polyphenolic compounds commonly found in Sauvignon blanc wine: catechin, caffeic acid and quercetin. Results showed each polyphenol had a unique effect when blended with a specific aroma compound, either suppressing, accentuating, or showing little effect on the perception of the aroma compounds. The perception of MIBP, 3MH, and ethyl decanoate were largely suppressed by the added polyphenols, with a few exceptions. The perception of 3MH was accentuated with the addition of caffeic acid, and the perception of 3MHA was accentuated with the addition of catechin. The interactive effects of aroma compounds with polyphenols likely reflect non-covalent associations in the wine solution that reduce the volatility of the aroma compounds. With an understanding of the interactive effects of volatile and non-volatile compounds in wine, winemakers might optimize the impact of selected volatile compounds by managing polyphenol levels, supporting their efforts to attain desirable wine aroma profiles.

The grape variety Sauvignon Blanc (SB) is the flagship of New Zealand’s wine industry and accounted for over 75 % of the value of total wine exports in 2008. Two volatile thiols, 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl-acetate (3MHA), reminiscent of grapefruit and passion fruit respectively, are critical for the main varietal characters in New Zealand SB. These aromatic thiols are not present in the grape juice, but are synthesized and released by the yeast during alcoholic fermentation from non-aromatic precursors. The aim of this work was to elucidate the underlying genetics of volatile thiol synthesis in yeast (Saccharomyces cerevisiae) during alcoholic fermentation of grape juice. A gene-deletion strategy was chosen for the investigation of putative genes influencing 3MH and 3MHA release. The first part of this thesis optimized fermentation conditions in grape-juice-based media, which enabled auxotrophic laboratory strains, derived from S288C, to ferment grape juice to completion with high efficiency. Key steps to achieving this goal were the supplementation of the grape juice with higher than recommended amounts of amino acids, which increased the fermentation rate of auxotrophic yeast strains. Lysine auxotrophic strains especially benefited from this measure. In combination with the dilution of SB grape juice by 25 % with synthetic grape juice without sugars, the auxotrophic laboratory yeast BY4743 was able to metabolize all sugars in the grape juice-based media in a time frame similar to that of a commercial wine yeast. The key properties of the resulting wine were comparable to wine made with a commercial wine yeast under the same conditions. In the second part, these newly developed fermentation conditions were employed to screen 69 single-gene deletion strains in the laboratory yeast background BY4743. The list of the 69 candidate genes was compiled by combining existing knowledge about thiol production in yeast with the mining of several biological databases. Screening of the single-gene deletions revealed 17 genes which caused biologically relevant increases or decreases in volatile thiol production, but none abolished it. The majority of the 17 genes were related to the sulfur and nitrogen metabolism in yeast. A subset of these thiol-influencing genes were also deleted in a wine yeast, and were overexpressed in both wine yeast and laboratory yeast, to gain more insight in their regulatory effects. The findings confirmed that sulfur and nitrogen metabolism in yeast were important in regulating 3MH and 3MHA synthesis. Different sulfur and nitrogen sources were added to the grape must prior to fermentation and their effect on thiol release was studied. It was found that nitrogen sources urea and DAP, as well as, the sulfur compound S-ethyl-L-cysteine (SEC) increased 3MH and 3MHA concentrations in the resulting wines. The addition of cysteine to grape juice fermented with wine yeast deleted in genes CYS3 and CYS4 more than doubled total thiol production. Mapping approaches to investigate thiol production in yeast were employed in the final part of this thesis. Genetically mapped F2 progeny of a cross between a low thiol-producing yeast strain and a high-thiol producer were screened for their thiol phenotype. The 3MH and 3MHA phenotypes across 48 screened F2 progeny resembled normal distributions, indicating a quantitative trait. Subsequent mapping identified a locus on chromosome 14 with a small effect on the 3MHA phenotype, but no obvious candidate genes were evident in the region. Another approach to investigate the evolution of volatile thiols in yeast included the use of SEC, a thiol compound resembling the cysteinylated precursor of 3MH, as a sole nitrogen source in a yeast growth assay. It was found that most wine yeast, European yeast isolates and laboratory yeasts could utilize SEC as a nitrogen source, whereas various other S. cerevisiae isolates could not. Crosses between three pairs of Sec- and Sec+ yeast strains strongly indicated that this trait was monogenically inherited. However, no direct correlation between the SEC phenotype and volatile release could be observed. Genetic mapping experiments in one SEC-segregating yeast population linked this SEC phenotype to the leu2-D0 deletion in a cross between a Leu+ and Leu- yeast strain. It was shown that leucine auxotrophy most likely caused the Sec- phenotype. In a second F2 population of a cross between prototrophic Sec+ and Sec- strains, strong linkage was established to a region on chromosome 6 containing two candidate genes, DUG1 and IRC7. DUG1 was proved not to be the cause of the SEC phenotype, whereas IRC7 remains a strong candidate gene.

ABSTRACT A core tool of sensory science is the use of trained descriptive panels. This research describes an investigation into the role of motivation in the performance of trained panels and the use of a trained panel to develop a better understanding of the perception of Sauvignon blanc wines. Substantial investment in time and money is directed towards ensuring trained panels perform optimally. Having selected a panel, the panel leader needs to ensure that panellists provide accurate, reliable data. Panellist motivation is also an important factor to consider. While performance psychology, education and sport science fields have researched motivation extensively, knowledge about panellist motivation within sensory science is limited. However, findings from existing research in these other areas - which suggest an important role for autonomy, competence and relatedness - can be applied to sensory panels in order to increase intrinsic motivation. The initial part of the research investigated the fundamental factors that affect and influence panellists’ motivation and participation. A survey (n=74) revealed that extra income and a general interest in food were the key drivers in inspiring people to become panellists, whilst enjoyment in being a panellist, interest in food, and extra income were key drivers for people to remain panellists. In a second survey, the intrinsic motivation of seven trained panels from four countries (n=108) was assessed. External panels were found to be more intrinsically motivated than internal panels. Experienced panellists had an increased perception of competence, which is a key factor for people to be intrinsically motivated. Understanding motivational frameworks currently used in other research fields and integrating them into existing panel training protocols may enhance and sustain panellists’ intrinsic motivation. A trained panel (n=14) was then used in the second part of the thesis to identify key flavours in Sauvignon blanc wines from Australia, France, New Zealand, Spain, South Africa and USA. Sixteen characteristics were identified and measured, including sweet sweaty passionfruit, capsicum, passionfruit skin/stalk, boxwood/cat’s urine, grassy, mineral/flinty, citrus, bourbon, apple lolly/candy, tropical, mint, fresh asparagus, canned asparagus, stonefruit, apple and snowpea. Principal component analysis was used to describe differences between regions and countries. Sauvignon blanc wines from Marlborough, New Zealand (NZ), were described by tropical and sweet sweaty passionfruit characteristics, while French and South African Sauvignon blanc wines were described as having flinty/mineral and bourbon-like flavors. Chemical analyses of these wines also showed that Marlborough, NZ wines had more methoxypyrazine and thiol compounds. A consumer study (n=109) showed that New Zealanders significantly prefer New Zealand style Sauvignon blanc. The final part of this research focused on using trained panellists to explore the interactions between volatile and non-volatile wine compounds and their effects on the aroma profile of New Zealand Sauvignon blanc wine. Four volatile aroma compounds that are important in New Zealand Sauvignon blanc wine were studied (isobutyl methoxypyrazine [MIBP], 3-mercaptohexanol [3MH], 3-mercaptohexanol acetate [3MHA], and ethyl decanoate). Each of these four aroma compounds were assessed in combination with three non-volatile polyphenolic compounds commonly found in Sauvignon blanc wine: catechin, caffeic acid and quercetin. Results showed each polyphenol had a unique effect when blended with a specific aroma compound, either suppressing, accentuating, or showing little effect on the perception of the aroma compounds. The perception of MIBP, 3MH, and ethyl decanoate were largely suppressed by the added polyphenols, with a few exceptions. The perception of 3MH was accentuated with the addition of caffeic acid, and the perception of 3MHA was accentuated with the addition of catechin. The interactive effects of aroma compounds with polyphenols likely reflect non-covalent associations in the wine solution that reduce the volatility of the aroma compounds. With an understanding of the interactive effects of volatile and non-volatile compounds in wine, winemakers might optimize the impact of selected volatile compounds by managing polyphenol levels, supporting their efforts to attain desirable wine aroma profiles.

The grape variety Sauvignon Blanc (SB) is the flagship of New Zealand’s wine industry and accounted for over 75 % of the value of total wine exports in 2008. Two volatile thiols, 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl-acetate (3MHA), reminiscent of grapefruit and passion fruit respectively, are critical for the main varietal characters in New Zealand SB. These aromatic thiols are not present in the grape juice, but are synthesized and released by the yeast during alcoholic fermentation from non-aromatic precursors. The aim of this work was to elucidate the underlying genetics of volatile thiol synthesis in yeast (Saccharomyces cerevisiae) during alcoholic fermentation of grape juice. A gene-deletion strategy was chosen for the investigation of putative genes influencing 3MH and 3MHA release. The first part of this thesis optimized fermentation conditions in grape-juice-based media, which enabled auxotrophic laboratory strains, derived from S288C, to ferment grape juice to completion with high efficiency. Key steps to achieving this goal were the supplementation of the grape juice with higher than recommended amounts of amino acids, which increased the fermentation rate of auxotrophic yeast strains. Lysine auxotrophic strains especially benefited from this measure. In combination with the dilution of SB grape juice by 25 % with synthetic grape juice without sugars, the auxotrophic laboratory yeast BY4743 was able to metabolize all sugars in the grape juice-based media in a time frame similar to that of a commercial wine yeast. The key properties of the resulting wine were comparable to wine made with a commercial wine yeast under the same conditions. In the second part, these newly developed fermentation conditions were employed to screen 69 single-gene deletion strains in the laboratory yeast background BY4743. The list of the 69 candidate genes was compiled by combining existing knowledge about thiol production in yeast with the mining of several biological databases. Screening of the single-gene deletions revealed 17 genes which caused biologically relevant increases or decreases in volatile thiol production, but none abolished it. The majority of the 17 genes were related to the sulfur and nitrogen metabolism in yeast. A subset of these thiol-influencing genes were also deleted in a wine yeast, and were overexpressed in both wine yeast and laboratory yeast, to gain more insight in their regulatory effects. The findings confirmed that sulfur and nitrogen metabolism in yeast were important in regulating 3MH and 3MHA synthesis. Different sulfur and nitrogen sources were added to the grape must prior to fermentation and their effect on thiol release was studied. It was found that nitrogen sources urea and DAP, as well as, the sulfur compound S-ethyl-L-cysteine (SEC) increased 3MH and 3MHA concentrations in the resulting wines. The addition of cysteine to grape juice fermented with wine yeast deleted in genes CYS3 and CYS4 more than doubled total thiol production. Mapping approaches to investigate thiol production in yeast were employed in the final part of this thesis. Genetically mapped F2 progeny of a cross between a low thiol-producing yeast strain and a high-thiol producer were screened for their thiol phenotype. The 3MH and 3MHA phenotypes across 48 screened F2 progeny resembled normal distributions, indicating a quantitative trait. Subsequent mapping identified a locus on chromosome 14 with a small effect on the 3MHA phenotype, but no obvious candidate genes were evident in the region. Another approach to investigate the evolution of volatile thiols in yeast included the use of SEC, a thiol compound resembling the cysteinylated precursor of 3MH, as a sole nitrogen source in a yeast growth assay. It was found that most wine yeast, European yeast isolates and laboratory yeasts could utilize SEC as a nitrogen source, whereas various other S. cerevisiae isolates could not. Crosses between three pairs of Sec- and Sec+ yeast strains strongly indicated that this trait was monogenically inherited. However, no direct correlation between the SEC phenotype and volatile release could be observed. Genetic mapping experiments in one SEC-segregating yeast population linked this SEC phenotype to the leu2-D0 deletion in a cross between a Leu+ and Leu- yeast strain. It was shown that leucine auxotrophy most likely caused the Sec- phenotype. In a second F2 population of a cross between prototrophic Sec+ and Sec- strains, strong linkage was established to a region on chromosome 6 containing two candidate genes, DUG1 and IRC7. DUG1 was proved not to be the cause of the SEC phenotype, whereas IRC7 remains a strong candidate gene.

ABSTRACT A core tool of sensory science is the use of trained descriptive panels. This research describes an investigation into the role of motivation in the performance of trained panels and the use of a trained panel to develop a better understanding of the perception of Sauvignon blanc wines. Substantial investment in time and money is directed towards ensuring trained panels perform optimally. Having selected a panel, the panel leader needs to ensure that panellists provide accurate, reliable data. Panellist motivation is also an important factor to consider. While performance psychology, education and sport science fields have researched motivation extensively, knowledge about panellist motivation within sensory science is limited. However, findings from existing research in these other areas - which suggest an important role for autonomy, competence and relatedness - can be applied to sensory panels in order to increase intrinsic motivation. The initial part of the research investigated the fundamental factors that affect and influence panellists’ motivation and participation. A survey (n=74) revealed that extra income and a general interest in food were the key drivers in inspiring people to become panellists, whilst enjoyment in being a panellist, interest in food, and extra income were key drivers for people to remain panellists. In a second survey, the intrinsic motivation of seven trained panels from four countries (n=108) was assessed. External panels were found to be more intrinsically motivated than internal panels. Experienced panellists had an increased perception of competence, which is a key factor for people to be intrinsically motivated. Understanding motivational frameworks currently used in other research fields and integrating them into existing panel training protocols may enhance and sustain panellists’ intrinsic motivation. A trained panel (n=14) was then used in the second part of the thesis to identify key flavours in Sauvignon blanc wines from Australia, France, New Zealand, Spain, South Africa and USA. Sixteen characteristics were identified and measured, including sweet sweaty passionfruit, capsicum, passionfruit skin/stalk, boxwood/cat’s urine, grassy, mineral/flinty, citrus, bourbon, apple lolly/candy, tropical, mint, fresh asparagus, canned asparagus, stonefruit, apple and snowpea. Principal component analysis was used to describe differences between regions and countries. Sauvignon blanc wines from Marlborough, New Zealand (NZ), were described by tropical and sweet sweaty passionfruit characteristics, while French and South African Sauvignon blanc wines were described as having flinty/mineral and bourbon-like flavors. Chemical analyses of these wines also showed that Marlborough, NZ wines had more methoxypyrazine and thiol compounds. A consumer study (n=109) showed that New Zealanders significantly prefer New Zealand style Sauvignon blanc. The final part of this research focused on using trained panellists to explore the interactions between volatile and non-volatile wine compounds and their effects on the aroma profile of New Zealand Sauvignon blanc wine. Four volatile aroma compounds that are important in New Zealand Sauvignon blanc wine were studied (isobutyl methoxypyrazine [MIBP], 3-mercaptohexanol [3MH], 3-mercaptohexanol acetate [3MHA], and ethyl decanoate). Each of these four aroma compounds were assessed in combination with three non-volatile polyphenolic compounds commonly found in Sauvignon blanc wine: catechin, caffeic acid and quercetin. Results showed each polyphenol had a unique effect when blended with a specific aroma compound, either suppressing, accentuating, or showing little effect on the perception of the aroma compounds. The perception of MIBP, 3MH, and ethyl decanoate were largely suppressed by the added polyphenols, with a few exceptions. The perception of 3MH was accentuated with the addition of caffeic acid, and the perception of 3MHA was accentuated with the addition of catechin. The interactive effects of aroma compounds with polyphenols likely reflect non-covalent associations in the wine solution that reduce the volatility of the aroma compounds. With an understanding of the interactive effects of volatile and non-volatile compounds in wine, winemakers might optimize the impact of selected volatile compounds by managing polyphenol levels, supporting their efforts to attain desirable wine aroma profiles.

The grape variety Sauvignon Blanc (SB) is the flagship of New Zealand’s wine industry and accounted for over 75 % of the value of total wine exports in 2008. Two volatile thiols, 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl-acetate (3MHA), reminiscent of grapefruit and passion fruit respectively, are critical for the main varietal characters in New Zealand SB. These aromatic thiols are not present in the grape juice, but are synthesized and released by the yeast during alcoholic fermentation from non-aromatic precursors. The aim of this work was to elucidate the underlying genetics of volatile thiol synthesis in yeast (Saccharomyces cerevisiae) during alcoholic fermentation of grape juice. A gene-deletion strategy was chosen for the investigation of putative genes influencing 3MH and 3MHA release. The first part of this thesis optimized fermentation conditions in grape-juice-based media, which enabled auxotrophic laboratory strains, derived from S288C, to ferment grape juice to completion with high efficiency. Key steps to achieving this goal were the supplementation of the grape juice with higher than recommended amounts of amino acids, which increased the fermentation rate of auxotrophic yeast strains. Lysine auxotrophic strains especially benefited from this measure. In combination with the dilution of SB grape juice by 25 % with synthetic grape juice without sugars, the auxotrophic laboratory yeast BY4743 was able to metabolize all sugars in the grape juice-based media in a time frame similar to that of a commercial wine yeast. The key properties of the resulting wine were comparable to wine made with a commercial wine yeast under the same conditions. In the second part, these newly developed fermentation conditions were employed to screen 69 single-gene deletion strains in the laboratory yeast background BY4743. The list of the 69 candidate genes was compiled by combining existing knowledge about thiol production in yeast with the mining of several biological databases. Screening of the single-gene deletions revealed 17 genes which caused biologically relevant increases or decreases in volatile thiol production, but none abolished it. The majority of the 17 genes were related to the sulfur and nitrogen metabolism in yeast. A subset of these thiol-influencing genes were also deleted in a wine yeast, and were overexpressed in both wine yeast and laboratory yeast, to gain more insight in their regulatory effects. The findings confirmed that sulfur and nitrogen metabolism in yeast were important in regulating 3MH and 3MHA synthesis. Different sulfur and nitrogen sources were added to the grape must prior to fermentation and their effect on thiol release was studied. It was found that nitrogen sources urea and DAP, as well as, the sulfur compound S-ethyl-L-cysteine (SEC) increased 3MH and 3MHA concentrations in the resulting wines. The addition of cysteine to grape juice fermented with wine yeast deleted in genes CYS3 and CYS4 more than doubled total thiol production. Mapping approaches to investigate thiol production in yeast were employed in the final part of this thesis. Genetically mapped F2 progeny of a cross between a low thiol-producing yeast strain and a high-thiol producer were screened for their thiol phenotype. The 3MH and 3MHA phenotypes across 48 screened F2 progeny resembled normal distributions, indicating a quantitative trait. Subsequent mapping identified a locus on chromosome 14 with a small effect on the 3MHA phenotype, but no obvious candidate genes were evident in the region. Another approach to investigate the evolution of volatile thiols in yeast included the use of SEC, a thiol compound resembling the cysteinylated precursor of 3MH, as a sole nitrogen source in a yeast growth assay. It was found that most wine yeast, European yeast isolates and laboratory yeasts could utilize SEC as a nitrogen source, whereas various other S. cerevisiae isolates could not. Crosses between three pairs of Sec- and Sec+ yeast strains strongly indicated that this trait was monogenically inherited. However, no direct correlation between the SEC phenotype and volatile release could be observed. Genetic mapping experiments in one SEC-segregating yeast population linked this SEC phenotype to the leu2-D0 deletion in a cross between a Leu+ and Leu- yeast strain. It was shown that leucine auxotrophy most likely caused the Sec- phenotype. In a second F2 population of a cross between prototrophic Sec+ and Sec- strains, strong linkage was established to a region on chromosome 6 containing two candidate genes, DUG1 and IRC7. DUG1 was proved not to be the cause of the SEC phenotype, whereas IRC7 remains a strong candidate gene.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias, Programa de Pós-Graduação em Ciência dos Alimentos, Florianópolis, 2010 / Made available in DSpace on 2012-10-25T07:12:03Z (GMT). No. of bitstreams: 1 282003.pdf: 1280843 bytes, checksum: e4f6203d2e606307f9144e2e9ecefadb (MD5) / A uva Cabernet Sauvignon, apresenta diferentes clones, que são classificados de acordo com seu potencial produtivo. Dentre estes clones pode-se destacar os clones 169 e 685, ambos de origem Francesa que são classificados em distintos grupos. O objetivo deste trabalho foi caracterizar dois clones, 169 e 685, da uva Cabernet Sauvignon procedentes da região de São Joaquim do Estado de Santa Catarina, Brasil, e validar uma metodologia por cromatografia líquida de alta eficiencia para a eterminação simultânea dos principais compostos fenólicos. Os vinhos produzidos com os clones 169 e 685, safra 2008, foram caracterizados quanto aos parâmetros físico-quimicos, compostos fenólicos totais, compostos fenólicos individuais flavonóides e não-flavonóides, ácidos orgânicos e atividade antioxidante. Também foi avaliado a evolução dos principais compostos fenólicos e da atividade antioxidante dos vinhos durante 11 meses de guarda em garrafa. As uvas dos clones 169 e 685, safra 2009, foram avaliadas quanto ao ciclo fenológico, evolução do período de maturação, componentes de rendimento e área foliar. Os resultados demonstraram que a metodologia desenvolvida para quantificar os principais compostos fenólicos presentes no vinho apresentou excelente precisão, exatidão, limites de detecção e recuperação (80-120 %), assim como demonstrou separação satisfatória de todos os compostos avaliados, podendo ser utilizada para diferenciar amostras de vinho. Os vinhos produzidos com os clones 169 e 685 apresentaram diferenças significativas em relação aos compostos fenólicos, sendo que através de analises multivariadas, como análise de Cluster e Análise de Componentes Principais, foi possível separar e classificar os vinhos de acordo com o clone. O clone 169 apresentou maior correlação com os polifenóis enquanto o clone 685 foi correlacionado com os parâmetros de cor, principalmente com as antocianinas, esta mesma correlação também foi observada para as amostras de uvas. As uvas dos clones 169 e 685 apresentaram semelhança na duração do ciclo fenológico e requerimento térmico (graus-dias), durante a safra 2009, apresentando boa maturação tecnológica e fenólica. Os clones apresentaram diferenças significativas em relação as componentes de rendimento, área foliar assim como no teor de polifenóis e antocianinas. Com base nos resultados obtidos pode-se concluir que os vinhos apresentaram as características particulares de acordo com o clone que foram produzidos.