The aim of the present study was to evaluate populational structures among fermenting S. cerevisiae populations. Analysis of six polymorphic microsatellite loci was performed in 468 S. cerevisiae isolates derived from a previous screening (using mtDNA RFLP or electrophoretic karyotyoing) of 2490 yeast strains obtained from spontaneous fermentations of grapes collected in three vineyards of the Vinho Verde Region (northwest Portugal), and one vineyard of the Languedoc Region (South France) during the 2001 – 2003 harvest seasons.  Among the 93 alleles obtained, 52 new alleles were identified. For all loci analyzed, observed heterozygosity was three to four times lower than the expected value, probably due to a strong populational substructuring. Populational structures were identified based on the accumulation of small allele-frequency differences across six loci in groups of strains. The present work is the first large-scale approach showing that microsatellite typing reveals a very fine population resolution of indigenous S. cerevisiae strains isolated from vineyards.This study was financially supported by the programs POCI 2010 (FEDER/FCT, POCTI/AGR/56102/2004) and AGRO (ENOSAFE, Nº 762)

Cambon, Brigitte

Casal, Margarida

Dequin, Sylvie

Valero, Eva

Universidade do Minho: RepositoriUM

Winery Year
Number of 
collected
samples
Number of 
spontaneous
fermentations
Number of
Isolates
Number of
Non-S. cerevisiae
isolates
12
6
12
12
3
12
12
9
12
12
12
90
12
103
6
6
8
1
7
8
5
10
11
1
12
0
0
0
0
0
0
0
0
129
359
1
2
2
2 2
2001 330 51 1
6 59
2002 360 1M
2003 180 21
2
180
180
240
30
210
240
150
300
Number of genotypes
2001 11 
34
41
26
1
35 
64 
12
59
A 2002
2003
2001
2002
2003
2001
2002P
2003
C
Abstract
The aim of the present study was to evaluate populational structures among 
fermenting S. cerevisiae populations. Analysis of six polymorphic microsatellite
loci was performed in 468 S. cerevisiae isolates derived from a previous screening 
(using mtDNA RFLP or electrophoretic karyotyoing) of 2490 yeast strains obtained 
from spontaneous fermentations of grapes collected in three vineyards of the 
Vinho Verde Region (northwest Portugal), and one vineyard of the Languedoc 
Region (South France) during the 2001 – 2003 harvest seasons.  Among the 93 
alleles obtained, 52 new alleles were identified. For all loci analyzed, observed 
heterozygosity was three to four times lower than the expected value, probably
due to a strong populational substructuring. Populational structures were 
identified based on the accumulation of small allele-frequency differences across 
six loci in groups of strains. The present work is the first large-scale approach 
showing that microsatellite typing reveals a very fine population resolution of 
indigenous S. cerevisiae strains isolated from vineyards.
References
[1] D. Schuller, H. Alves, S. Dequin, M. Casal. 2005. FEMS Microbiol Ecol 51, 167-177.
[2] E. Valero, D. Schuller, B. Cambon, M. Casal, S. Dequin. 2005. FEMS Yeast Res. 5, 959-969.
[3] Pretorius IS, du Toit M, van Rensburg P (2003). Food Technol Biotechnol 41:3-10.
[4] Barnett J. A., Payne, R. W., and Yarrow, D. 1990. Yeasts: Characteristics and identification. 2nd 
Ed. Cambridge University Press. Cambridge. UK.
[5] Blondin, B., and Vezinhet, F. 1988. Rev. Fr. Oenol. 28: 7-11.
[6] Pérez, M.A., Gallego, F.J., Martinez, I. and Hidalgo, P. 2001. Lett. Appl. Microbiol. 33, 461-466.
[7] López V., Querol A., Ramón D., and Fernández-Espinar M.T. 2001. Int J Food Microbiol 68:75-
81
[8] Schuller, D., Valero, E., Dequin, S. and Casal, M. 2004. FEMS Microbiol Lett. 231(1) 19-26.
[9] Schneider, S., Kueffer, J-M., Roessli, D. and Excoffier, L. 1997. ARLEQUIN version 1.1. 
Software for population genetics data analysis. Geneva, Switzerland
[10] NTSYSpc 2.0, 1997, Exeter Software (http://www.ExeterSoftware.com)
Dorit Schuller1, Eva Valero2,3, Brigitte Cambon2, Sylvie Dequin2, Margarida Casal1
1 Departamento / Centro de Biologia, Universidade do Minho, Braga, Portugal 
2 UMR Sciences pour l’Oenologie, INRA, Montpellier, France
3 Present address: Departamento de Agroalimentación, IMIDRA, Madrid, Spain
Conclusions
Microsatellite typing of loci ScAAT1-ScAAT6, followed by statistical analysis permitted a high resolution populational screen, and
is therefore the appropriate method to obtain a deeper insight in the ecology and biogeography of fermentative S. cerevisiae
strains, even among geographically close regions.
Genetic differences among S. cerevisiae populations derived mainly from gradations in allele frequencies rather than from
distinctive “diagnostic” genotypes, and the accumulation of small allele-frequency differences across six loci allowed the
identification of a population structure. 
The extension of the current approach to strains isolated from other viticultural regions is currently underway, since a preliminary
comparison revealed major differences in both allelic combinations and frequencies (our unpublished data). 
Materials and Methods
Computer assisted data 
analysis
A group of strains with unique microsatellite profiles
(obtained from 30 isolates per fermentation) was considered
the population corresponding to each sampling site. The
pattern and degree of temporal and spatial divergence in the
nuclear microsatellites ScAAT1 to ScAAT6 among
subpopulations was estimated by Fst determination over all
loci by AMOVA analysis (computed by the Arlequin software 
[9]). A similarity matrix of allelic frequencies was computed 
by the program NTSYSpc 2.0 [10], based on the Euclidean 
distance and average linkage (UPGMA). 
Dorit Schuller
Centro de Biologia, 
Campus de Gualtar
Universidade do Minho
4710-057 Braga, Portugal  
Tel: 253 - 60 40 10/17
Fax: 253 – 67 89 80
Mail: dschuller@bio.uminho.pt
Comparison of vineyard-associated Saccharomyces
cerevisiae populations by microsatellite analysis
Samples
The sampling plan included four vineyards in the North of Portugal (Região Demarcada dos 
Vinhos Verdes), and South of France (Languedoc) as shown. In each vineyard, six sampling 
points were defined. Sampling campaigns were performed in duplicate (some days before and 
some days after harvest). The yeast flora from fermenting grape juice (500 ml) was analysed when 
the must weight was reduced by 70 g/l, corresponding to the consumption of about 2/3 of the 
sugar content. This experiment was repeated in three consecutive years (2001-2003), resulting in 
a total of 83 grape samples and 2490 isolates, that were further analyzed by molecular methods.
Introduction
The grape’s yeast flora depends on a large variety of factors such as climatic conditions 
including temperature and rainfalls, the geographic localization of the vineyard, antifungal 
applications, the harvest technique, grape variety, the vineyard’s age as well as the soil type. 
Several ecological surveys report a large diversity of Saccharomyces sp. strains among the 
enological fermentative flora. Some strains seem to be widely distributed in a given viticultural
region, can be found in several consecutive years and are also predominant in the fermenting 
flora hypothesizing the occurrence of specific native strains that can be associated to a terroir
[1-2]. 
At  present, leading winemakers demand for autochthonous fermenting strains that are able to 
enhance the expression of typical sensorial characteristics of wine and ensure the control of the 
fermentation process, concerning the motto “special yeasts for special traits” [3]. The detailed 
biogeographical evaluation of fermentative strains is essential for the establishment of adequate 
selection and improvement programes. 
The aim of the present study was to gain insight in the populational structure of vineyard-
associated S. cerevisiae in vineyards located in South France and North Portugal. This is the first 
systematical, 3-years biogeographical survey of fermentative S. cerevisiae strains by 
microsatellite genotyping, aiming at the analysis of populational structures and genetic
variability in three vineyards of the Vinho Verde Wine Region of Portugal. 
Number of perennial genotypes (regional distribution)
1 Number of perennial genotypes (limited to one vinayard)
Number of annual genotypes (multiple sites of one vineyard)
1 Number of annual genotypes (in multiple sites of two vineyards)
This poster is available at
http://repositorium.sdum.uminho.pt
The six markers revealed a high degree of genetic variability, being ScAAT1 and ScAAT3 the most polymorphic markers with 31 and 19 
alleles, respectively.
Besides the 41 ScAAT1-ScAAT6 alleles previously described for 51 strains [3], 52 new alleles were identified in the present study .
Some newly described alleles occur with relative high frequency and may be used as indicative alleles for the Vinho Verde Wine
Region. 
The vast majority of alleles were evenly distributed among S. cerevisiae populations belonging to vineyards A, C and P and M, but
differences are notorious for few alleles, which can be considered as vineyard(s) or Wine Region – indicative . 
Acknowledgements
This study was supported by the projects ENOSAFE (Nº 762, Programa AGRO, medida 8) and the 
programme POCI 2010 (project POCI/AGR/56771/2004). We appreciate the kind assistance of the
enologists Rui Cunha, Anselmo Mendes, Euclides Rodrigues and José Domingues for facilitating
sampling campaigns in the three vineyards. Magda Silva Graça is gratefully acknowledged for the
operation of the DNA sequencer. 
Relationships among the populations from different 
sampling sites in four vineyards 
R
E
S
U
L
T
S
Allelic frequenciesStrains collected
The strain collection obtained from this survey comprises 2490 isolates, that were classified in 356 genetic
patterns according their allelic distribution. 
The highest S. cerevisiae biodiversity was observed in winery M (323 isolates, 73 patterns) followed by
wineries P (690 isolates, 135 patterns), A (450 isolates, 86 patterns) and C (480 isolates, 62 patterns). 
Several genotypes showed a wider temporal and geographical generalized pattern of sporadic presence, 
absence and reappearance across sampling sites, vineyards or years. 
Non-Saccharomyces strains belonging to the genus Kloeckera.
Vineyard P
3
10 m
4
5
6
2 1
W
3
1
2
4
65
W
100 m
Vineyard C
Vineyard A
100 mW
6
5
4
3 2 1
Vineyard M
1000 m 6
5
4
3 2 1
W
WineryW
Molecular identification
In France
Preliminary discrimination between Saccharomyces and non-
Saccharomyces yeast was based on the inability to grow in YNB medium
containing L-lysine [4]. Chromosomal polymorphisms were studied by 
pulsed field gel electrophoresis as previously described [5]. Strains with 
identical mtDNA RFLP patterns were grouped and one representative 
strain was further characterised by analysis of 6  microsatellite loci [6].
In Portugal
Isolated strains were analysed by mitochondrial DNA restriction patterns 
(mtDNA RFLP) [7]. Strains with identical mtDNA RFLP patterns were 
grouped and one representative strain was further characterised by 
analysis of the above mentioned  microsatellite loci.The equivalence of this 
typing method to previously described ones has been previously shown
[8]. Microsatellite loci are specific for Saccharomyces cerevisiae. Other
Saccharomyces species that may be present in spontaneous fermentations
such as S. bayanus and S.paradoxus showed no amplification signals in 
most of the loci. 
(AAT)19IXScAAT6
(AAT)13XVIScAAT5
(ATT)12VIIScAAT4
(ATT)23IVScAAT3
(ATT)20IIScAAT2
(ATT)35XIIIScAAT1
Repeat(number) ChromosomeMicrosatellite
0
0,2
0,4
0,6
0,8
1
1,2
1,4
1,6
156 159 165 171 174 177 180 186 189 192 195 198 201 204 207 213 216 219 222 225 228 237 240 243 249 261 267 273 279 282 300
0
0,4
0,8
1,2
1,6
2
2,4
2,8
287 293 299 302 305 311 314 317 323 326 329 332 335 338 341 347 350
0
0,2
0,4
0,6
0,8
1
1,2
1,4
339 345 348 351 354 357 360 363 366 369 372 375 378 381 384
0
0,2
0,4
0,6
0,8
1
1,2
1,4
1,6
213 216 219 222 225
0
0,2
0,4
0,6
0,8
1
1,2
1,4
229 235 238 241 244 247 250 253 256 259 262 265 268 271 274 277 280 301 346
0
0,4
0,8
1,2
1,6
2
2,4
2,8
3,2
247 250 253 256 259 262 265 271 277
ScAAT1
ScAAT6ScAAT3ScAAT5ScAAT2
ScAAT4
Al
lel
ic
fre
qu
en
cy
Al
lel
ic
Al
lel
ic
fre
qu
en
cy
fre
qu
en
cy
Allele (bp)
Vineyard M
Vineyard C
Vineyard P
Vineyard A
AMOVA analysis - Fst values based on microsatellite data 
Coefficient
0.45 0.65 0.85 1.05 1.25
          
 A1(20) 
 A3(26) 
 A4(17) 
 C2(26) 
 P3(17) 
 P1(35) 
 P2(20) 
 P6(20) 
 P4(38) 
 P5(29) 
 C4(23) 
 C5(15) 
 M2(16) 
 M4(14) 
 M5(15) 
 M3(29) 
 M6(12) 
 A5(29) 
Vineyard-specific populational substructure is shown by several clusters, comprising sampling sites of vineyards C, 
P, A and M. Populations within groups C and P are more closely related, while S. cerevisiae populations belonging
to vineyard A are much more heterogeneous and also more distinct from C and P. 
The C2 population lies within the P-cluster, indicating that genetic differences do not delimit specific populations
with fixed geographic boundaries. 
Strains from Languedoc form a cluster that can be clearly distinguished from strains isolated in the Vinho Verde 
Region
Exceptions from the vineyard - specific population structure may be due to the presence of rare alleles (A5).
r = 0.85
The numbers of strains with distinct
genotypes are indicated in parenthesis
0.00010.0793.336.300.37P
0.070.0594.615.63-0.25M
0.00010.1188.8912.67-1.56C
< 0.00010.1882.0520.48-2.45A
2001 / 2003
0.0520.0990.597.731.68P
< 0.00010.1783.015.791.29A
2002 / 2003
0.00010.1189.279.940.79P
0.0340.1188.5113.94-2.45A
2001 / 2002
Among years
< 0.00010.0891.984.543.482003
< 0.00010.0891.805.442.75M/P2001
0.0010.1386.909.253.852003
0.0160.0694.255.720.03
M/C
2001
< 0.00010.2278.0414.597.382003
< 0.00010.1386.807.335.87
M/A
2001
< 0.00010.0991.428.100.482003
0.00010.0792.898.31-1.21C/P2001
< 0.00010.1782.7116.201.092003
0.0590.1287.5116.66-4.16
A/C
2001
0.00010.1485.9511.292.762003
0.00010.2080,3313.286.382002
< 0.00010.1287.949.033.03
A/P
2001
Among vineyards
WPAPWGAG P(r < o)FSTSource of variation
WP  within population (group of genotypes from
duplicate campaigns of one sampling site);
APWG among populations within groups of strains
from different sampling years / vineyards
AG among groups of strains from different
sampling years / vineyards
Genetic distance, expressed as Fst values, are 
not correlated with the distance between 
vineyards:
Pair wise association of populations from 
different vineyards showed that the closer
vineyards A/P and A/C (30 – 50 km) are 
genetically less related compared to the more 
distant vineyards P/C (ca. 80 km). 
Fst values for the pair wise association of S. 
cerevisiae populations from France and Portugal 
(A/M, C/M and P/M) are similar to the values
observed among Portuguese populations.
Populations variation within a vineyard in 
consecutive years is similar to the variation
observed between vineyards, being more 
variable in A (Fst = 0.11 – 0.18) compared to P 
(Fst = 0.09 – 0.11). 


English

Comparison of vineyard-associated Saccharomyces cerevisiae populations by microsatellite analysis

Winery YearNumber of collectedsamplesNumber of spontaneousfermentationsNumber ofIsolatesNumber ofNon-S. cerevisiaeisolates1261212312129121212901210366817851011Y112000000001293591222 22001 330 51 1[Y6 592002 360 1M2003 180 21XYZX2XXXXX18018024030210240150300Number of genotypes2001 11 344126135 64 1259A 2002200320012002200320012002P2003CAbstractThe aim of the present study was to evaluate populational structures among fermenting S. cerevisiae populations. Analysis of six polymorphic microsatelliteloci was performed in 468 S. cerevisiae isolates derived from a previous screening (using mtDNA RFLP or electrophoretic karyotyoing) of 2490 yeast strains obtained from spontaneous fermentations of grapes collected in three vineyards of the Vinho Verde Region (northwest Portugal), and one vineyard of the Languedoc Region (South France) during the 2001 – 2003 harvest seasons.  Among the 93 alleles obtained, 52 new alleles were identified. For all loci analyzed, observed heterozygosity was three to four times lower than the expected value, probablydue to a strong populational substructuring. Populational structures were identified based on the accumulation of small allele-frequency differences across six loci in groups of strains. The present work is the first large-scale approach showing that microsatellite typing reveals a very fine population resolution of indigenous S. cerevisiae strains isolated from vineyards.References[1] D. Schuller, H. Alves, S. Dequin, M. Casal. 2005. FEMS Microbiol Ecol 51, 167-177.[2] E. Valero, D. Schuller, B. Cambon, M. Casal, S. Dequin. 2005. FEMS Yeast Res. 5, 959-969.[3] Pretorius IS, du Toit M, van Rensburg P (2003). Food Technol Biotechnol 41:3-10.[4] Barnett J. A., Payne, R. W., and Yarrow, D. 1990. Yeasts: Characteristics and identification. 2nd Ed. Cambridge University Press. Cambridge. UK.[5] Blondin, B., and Vezinhet, F. 1988. Rev. Fr. Oenol. 28: 7-11.[6] Pérez, M.A., Gallego, F.J., Martinez, I. and Hidalgo, P. 2001. Lett. Appl. Microbiol. 33, 461-466.[7] López V., Querol A., Ramón D., and Fernández-Espinar M.T. 2001. Int J Food Microbiol 68:75-81[8] Schuller, D., Valero, E., Dequin, S. and Casal, M. 2004. FEMS Microbiol Lett. 231(1) 19-26.[9] Schneider, S., Kueffer, J-M., Roessli, D. and Excoffier, L. 1997. ARLEQUIN version 1.1. Software for population genetics data analysis. Geneva, Switzerland[10] NTSYSpc 2.0, 1997, Exeter Software (http://www.ExeterSoftware.com)Dorit Schuller1, Eva Valero2,3, Brigitte Cambon2, Sylvie Dequin2, Margarida Casal11 Departamento / Centro de Biologia, Universidade do Minho, Braga, Portugal 2 UMR Sciences pour l’Oenologie, INRA, Montpellier, France3 Present address: Departamento de Agroalimentación, IMIDRA, Madrid, SpainConclusionsMicrosatellite typing of loci ScAAT1-ScAAT6, followed by statistical analysis permitted a high resolution populational screen, andis therefore the appropriate method to obtain a deeper insight in the ecology and biogeography of fermentative S. cerevisiaestrains, even among geographically close regions.Genetic differences among S. cerevisiae populations derived mainly from gradations in allele frequencies rather than fromdistinctive “diagnostic” genotypes, and the accumulation of small allele-frequency differences across six loci allowed theidentification of a population structure. The extension of the current approach to strains isolated from other viticultural regions is currently underway, since a preliminarycomparison revealed major differences in both allelic combinations and frequencies (our unpublished data). Materials and MethodsComputer assisted data analysisA group of strains with unique microsatellite profiles(obtained from 30 isolates per fermentation) was consideredthe population corresponding to each sampling site. Thepattern and degree of temporal and spatial divergence in thenuclear microsatellites ScAAT1 to ScAAT6 amongsubpopulations was estimated by Fst determination over allloci by AMOVA analysis (computed by the Arlequin software [9]). A similarity matrix of allelic frequencies was computed by the program NTSYSpc 2.0 [10], based on the Euclidean distance and average linkage (UPGMA). Dorit SchullerCentro de Biologia, Campus de GualtarUniversidade do Minho4710-057 Braga, Portugal  Tel: 253 - 60 40 10/17Fax: 253 – 67 89 80Mail: dschuller@bio.uminho.ptComparison of vineyard-associated Saccharomycescerevisiae populations by microsatellite analysisSamplesThe sampling plan included four vineyards in the North of Portugal (Região Demarcada dos Vinhos Verdes), and South of France (Languedoc) as shown. In each vineyard, six sampling points were defined. Sampling campaigns were performed in duplicate (some days before and some days after harvest). The yeast flora from fermenting grape juice (500 ml) was analysed when the must weight was reduced by 70 g/l, corresponding to the consumption of about 2/3 of the sugar content. This experiment was repeated in three consecutive years (2001-2003), resulting in a total of 83 grape samples and 2490 isolates, that were further analyzed by molecular methods.IntroductionThe grape’s yeast flora depends on a large variety of factors such as climatic conditions including temperature and rainfalls, the geographic localization of the vineyard, antifungal applications, the harvest technique, grape variety, the vineyard’s age as well as the soil type. Several ecological surveys report a large diversity of Saccharomyces sp. strains among the enological fermentative flora. Some strains seem to be widely distributed in a given viticulturalregion, can be found in several consecutive years and are also predominant in the fermenting flora hypothesizing the occurrence of specific native strains that can be associated to a terroir[1-2]. At  present, leading winemakers demand for autochthonous fermenting strains that are able to enhance the expression of typical sensorial characteristics of wine and ensure the control of the fermentation process, concerning the motto “special yeasts for special traits” [3]. The detailed biogeographical evaluation of fermentative strains is essential for the establishment of adequate selection and improvement programes. The aim of the present study was to gain insight in the populational structure of vineyard-associated S. cerevisiae in vineyards located in South France and North Portugal. This is the first systematical, 3-years biogeographical survey of fermentative S. cerevisiae strains by microsatellite genotyping, aiming at the analysis of populational structures and geneticvariability in three vineyards of the Vinho Verde Wine Region of Portugal. X Number of perennial genotypes (regional distribution)1 Number of perennial genotypes (limited to one vinayard)X Number of annual genotypes (multiple sites of one vineyard)1 Number of annual genotypes (in multiple sites of two vineyards)This poster is available athttp://repositorium.sdum.uminho.ptThe six markers revealed a high degree of genetic variability, being ScAAT1 and ScAAT3 the most polymorphic markers with 31 and 19 alleles, respectively.Besides the 41 ScAAT1-ScAAT6 alleles previously described for 51 strains [3], 52 new alleles were identified in the present study .Some newly described alleles occur with relative high frequency and may be used as indicative alleles for the Vinho Verde WineRegion. The vast majority of alleles were evenly distributed among S. cerevisiae populations belonging to vineyards A, C and P and M, butdifferences are notorious for few alleles, which can be considered as vineyard(s) or Wine Region – indicative . AcknowledgementsThis study was supported by the projects ENOSAFE (Nº 762, Programa AGRO, medida 8) and the programme POCI 2010 (project POCI/AGR/56771/2004). We appreciate the kind assistance of theenologists Rui Cunha, Anselmo Mendes, Euclides Rodrigues and José Domingues for facilitatingsampling campaigns in the three vineyards. Magda Silva Graça is gratefully acknowledged for theoperation of the DNA sequencer. Relationships among the populations from different sampling sites in four vineyards RESULTSAllelic frequenciesStrains collectedThe strain collection obtained from this survey comprises 2490 isolates, that were classified in 356 geneticpatterns according their allelic distribution. The highest S. cerevisiae biodiversity was observed in winery M (323 isolates, 73 patterns) followed bywineries P (690 isolates, 135 patterns), A (450 isolates, 86 patterns) and C (480 isolates, 62 patterns). Several genotypes showed a wider temporal and geographical generalized pattern of sporadic presence, absence and reappearance across sampling sites, vineyards or years. Non-Saccharomyces strains belonging to the genus Kloeckera.Vineyard P310 m4562 1W312465W100 mVineyard CVineyard A100 mW6543 2 1Vineyard M1000 m 6543 2 1WWineryWMolecular identificationIn FrancePreliminary discrimination between Saccharomyces and non-Saccharomyces yeast was based on the inability to grow in YNB mediumcontaining L-lysine [4]. Chromosomal polymorphisms were studied by pulsed field gel electrophoresis as previously described [5]. Strains with identical mtDNA RFLP patterns were grouped and one representative strain was further characterised by analysis of 6  microsatellite loci [6].In PortugalIsolated strains were analysed by mitochondrial DNA restriction patterns (mtDNA RFLP) [7]. Strains with identical mtDNA RFLP patterns were grouped and one representative strain was further characterised by analysis of the above mentioned  microsatellite loci.The equivalence of this typing method to previously described ones has been previously shown[8]. Microsatellite loci are specific for Saccharomyces cerevisiae. OtherSaccharomyces species that may be present in spontaneous fermentationssuch as S. bayanus and S.paradoxus showed no amplification signals in most of the loci. (AAT)19IXScAAT6(AAT)13XVIScAAT5(ATT)12VIIScAAT4(ATT)23IVScAAT3(ATT)20IIScAAT2(ATT)35XIIIScAAT1Repeat(number) ChromosomeMicrosatellite00,20,40,60,811,21,41,6156 159 165 171 174 177 180 186 189 192 195 198 201 204 207 213 216 219 222 225 228 237 240 243 249 261 267 273 279 282 30000,40,81,21,622,42,8287 293 299 302 305 311 314 317 323 326 329 332 335 338 341 347 35000,20,40,60,811,21,4339 345 348 351 354 357 360 363 366 369 372 375 378 381 38400,20,40,60,811,21,41,6213 216 219 222 22500,20,40,60,811,21,4229 235 238 241 244 247 250 253 256 259 262 265 268 271 274 277 280 301 34600,40,81,21,622,42,83,2247 250 253 256 259 262 265 271 277ScAAT1ScAAT6ScAAT3ScAAT5ScAAT2ScAAT4AllelicfrequencyAllelicAllelicfrequencyfrequencyAllele (bp)Vineyard MVineyard CVineyard PVineyard AAMOVA analysis - Fst values based on microsatellite data Coefficient0.45 0.65 0.85 1.05 1.25           A1(20)  A3(26)  A4(17)  C2(26)  P3(17)  P1(35)  P2(20)  P6(20)  P4(38)  P5(29)  C4(23)  C5(15)  M2(16)  M4(14)  M5(15)  M3(29)  M6(12)  A5(29) Vineyard-specific populational substructure is shown by several clusters, comprising sampling sites of vineyards C, P, A and M. Populations within groups C and P are more closely related, while S. cerevisiae populations belongingto vineyard A are much more heterogeneous and also more distinct from C and P. The C2 population lies within the P-cluster, indicating that genetic differences do not delimit specific populationswith fixed geographic boundaries. Strains from Languedoc form a cluster that can be clearly distinguished from strains isolated in the Vinho Verde RegionExceptions from the vineyard - specific population structure may be due to the presence of rare alleles (A5).r = 0.85The numbers of strains with distinctgenotypes are indicated in parenthesis0.00010.0793.336.300.37P0.070.0594.615.63-0.25M0.00010.1188.8912.67-1.56C< 0.00010.1882.0520.48-2.45A2001 / 20030.0520.0990.597.731.68P< 0.00010.1783.015.791.29A2002 / 20030.00010.1189.279.940.79P0.0340.1188.5113.94-2.45A2001 / 2002Among years< 0.00010.0891.984.543.482003< 0.00010.0891.805.442.75M/P20010.0010.1386.909.253.8520030.0160.0694.255.720.03M/C2001< 0.00010.2278.0414.597.382003< 0.00010.1386.807.335.87M/A2001< 0.00010.0991.428.100.4820030.00010.0792.898.31-1.21C/P2001< 0.00010.1782.7116.201.0920030.0590.1287.5116.66-4.16A/C20010.00010.1485.9511.292.7620030.00010.2080,3313.286.382002< 0.00010.1287.949.033.03A/P2001Among vineyardsWPAPWGAG P(r < o)FSTSource of variationWP  within population (group of genotypes fromduplicate campaigns of one sampling site);APWG among populations within groups of strainsfrom different sampling years / vineyardsAG among groups of strains from differentsampling years / vineyardsGenetic distance, expressed as Fst values, are not correlated with the distance between vineyards:Pair wise association of populations from different vineyards showed that the closervineyards A/P and A/C (30 – 50 km) are genetically less related compared to the more distant vineyards P/C (ca. 80 km). Fst values for the pair wise association of S. cerevisiae populations from France and Portugal (A/M, C/M and P/M) are similar to the valuesobserved among Portuguese populations.Populations variation within a vineyard in consecutive years is similar to the variationobserved between vineyards, being more variable in A (Fst = 0.11 – 0.18) compared to P (Fst = 0.09 – 0.11). 

http://repositorium.sdum.uminho.pt/bitstream/1822/6137/1/2006-FEMS-Genetic%20structure-Poster.pdf

Comparison of vineyard-associated Saccharomyces cerevisiae populations by microsatellite analysis

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Universidade do Minho: RepositoriUM

Universidade do Minho: RepositoriUM

Universidade do Minho: RepositoriUM