Genetic and Physiological Characterization of Brettanomyces bruxellensis Strains Isolated from Wines

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Figure 1
Phylogenetic tree obtained by alignment of the sequenced portion of 26S rDNA (Arg.: Argentina; Bel.: Belgium; New Z.: New Zealand; Port.: Portugal; MO: Missouri; TS: type strain sequence).
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Figure 2
Cluster analysis of the physiological-genetic data matrix. Text boxes are highlighting the fact that the defined groups are characterized by a different production of ethyl phenols. Low: <100 μg/L (13 strains); medium (in bold font): ~1000 μg/L (6 strains); high: ~2000 μg/ L (16 strains).
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Figure 3
Predictions on the first two discriminant functions calculated based on the 26S rDNA physiological-genetic profile. Groups are enclosed by their respective convex hulls. The first two linear discriminants account for 63% and 31% of the trace (between-group variance), respectively.

Table 1

Brettanomyces isolates, collections, origin, variety, and vintage.

UCD^a	NYSAES^a	Other collection^a	Geographic origin	Variety^b	Vintage
^aUCD: UC Davis Wine Yeast and Bacteria Collection; NYSAES: New York State Agricultural Experiment Station; CBS: Centraalbureau voor Shimmelcultures; UCD FST: UCD Food Science and Technology; DBVPG: Dipartmento di Biologia Vegetale Università di Perugia.
^bCH: Chardonnay; CS: Cabernet Sauvignon; M: Merlot; PN: Pinot noir; Z: Zinfandel; na: not available.
615			California	na	1974
738			Missouri	na	1982
752			France	na	1984
2030			California	CS	1972
2041			Thailand	Fruit	2002
2046			California	M	1999
2047			California	CS	2001
2048			California	PN	1999
2049			New Zealand	PN	2001
2050			New Zealand	CH	2001
2051			California	PN	2000
2052			California	Z	2001
2053			California	CS	2001
2054			California	CS	2001
2058		CBS73, UCD FST82-30	France	Must	1933
2059		UCD FST77-105	California	CS	1975
2060			Chile	CS	2001
2062		UCD FST75-41	France	Champagne	1975
2063		UCD FST77-106	California	CS	1975
2065		DBVPG4051	Malta	White must	na
2066		DBVPG4052	Malta	White must	na
2067			California	PN	2000
2075			California	na	2001
2076			Canada, BC	na	2001
2077			Chile	CS	2001
2078	CE40	CBS 72	Belgium	Beer	1931
2079	CE41	CBS 74	Belgium	Beer	1940
2080	CE116		Germany	na	na
2081	CE119		California	CS	1989
2082	CE120		California	CS	1989
2083	CE122		California	CS	1992
2085	CE50		California	na	na
2091		CBS 2797	France	Bordeaux	1957
2092		CBS 2796	Germany	Sparkling Moselle	1957
2093			New York	na	2001
	CE121		California	na	na
	CE148		Portugal	na	na
	CE149		Canada	na	na
	CE150		Canada	na	na
	HKY6		New Zealand	na	na
	HKY 8		Chile	na	na
	HKY65		Italy	na	na
	HKY99		New York	na	na
	HKY23		Portugal	na	na
	HKY33		Portugal	na	na
	HKY14		Argentina	na	na
	HKY47		Italy	na	na
	HKY86		Italy	na	na

Table 2

Summary of physiological characteristics of 35 Brettanomyces strains.

Character tested	Frequency	Isolates (%)
Carbon source growth
Arginine, cellulose, proline, tartrate	0	0
Adonitol	2	6
Arabinose, citrate, starch	3	9
Lactose, mannitol, raffinose	4	11
Ethanol	9	26
Glycerol	10	29
Lactate	12	34
Succinate	13	37
Malate	14	40
Galactose	28	80
Cellobiose, maltose	32	91
Trehalose	34	97
Sucrose	35	100
Nitrogen source growth
Nitrate	25	71
Arginine, proline	35	100
Temperature growth
at 37°C	13	37
at 10°C	11	31
Alcohol
Tolerance >10%	35	100
Sulfite tolerance
>30 mg/L at pH 3.4	17	49
pH growth
at pH 2.0	33	94
4-EP and 4-EG (μg/L)
High (>2000 4-EP; >1500 4-EG)	17	49
Medium (1000–2000 4-EP;700–1500 4-EG)	6	17
Low (<50 4-EP; <60 4-EG)	7	20
None (<4.0 4-EP and 4-EG)	7	17

Table 3

26S rDNA genetic profile compared to the strain origin, SO₂ tolerance, growth on ethanol and at 10°C, and volatile phenol production.

Strain	Growth at 10°C	SO₂ tolerance (mg/L)	4-EP (μg/L)	4-EG (μg/L)	Growth on ethanol	Origin	26S rDNA group
^ana: data not available.
752	++	20	32	18	+	France	na^a
2058	+	20	12	24	+	France	a
2063	−	30	1820	1330	−	California	a
2079	−	50	26	17	−	Belgium	a
2092	+	50	47	34	+	Germany	a
2093	+	50	2800	2060	+	New York	a
2091	+	20	0	0	+	France	a
2078	−	10	0	0	−	Belgium	a
2030	−	20	10	7	−	California	b
2048	−	50	2720	2450	−	California	b
2059	−	50	2810	2310	−	California	b
2067	−	30	1680	1230	−	California	b
2076	−	50	2870	2310	−	Canada	b
2077	−	50	2830	1930	−	Chile	b
2081	+	50	20	56	−	California	b
2082	−	50	2760	2030	−	California	b
2083	−	50	2860	2340	−	California	b
2085	−	50	2770	2190	−	California	b
2041	−	10	0	0	−	Thailand	b
2053	−	20	1564	1000	−	California	b
2062	−	30	2610	2090	−	France	b
2075	−	30	2810	2220	−	California	b
2060	−	30	2800	2340	−	Chile	b
615	+	20	2600	2130	+	California	c
738	++	30	2680	2380	−	Missouri	c
2047	+	10	1880	1340	+	California	c
2049	+	10	1113	870	−	New Zealand	c
2050	+	20	2470	1860	−	New Zealand	c
2054	+	20	1261	790	+	California	c
2051	+	20	2350	1720	−	California	c
2052	+	10	2590	1860	−	California	c
2046	−	20	0	0	−	California	d
2065	−	10	4	5	−	Malta	d
2066	+	10	0	0	+	Malta	d
2080	−	10	0	0	−	Germany	e