Research Findings That Will Change the Way You Make Wine

Research Findings That Will Change the Way You Make Wine Curtis Phillips, Moderator Wine Business Monthly David Block UC Davis Dan Durall University of British Columbia Sydney Morgan, Ph.D. University of British Columbia

Curtis Phillips - Moderator Senior Technical Editor Wine Business Monthly

David Block Marvin Sands Department Chair Viticulture and Enology UC Davis

Dan Durall Associate Professor, University of British Columbia

Yeasts in Winery Fermentations Five Years of Sampling Daniel Durall, Jessica Lange, Erin Faasse, Marissa Neuner, Chrystal Scholl, Amy Kluftinger, Sydney Morgan, Morgan Stone, Mansak Tantikachornkiat

Okanagan Valley

Types of Fermentation Inoculated (ADY) Spontaneous

Inoculated Fermentation What are we trying to achieve with an inoculated fermentation? Pure inoculum Consistency from one vintage to the next A predictable aroma and flavour of the wine Do we achieve any of these when conducting inoculated fermentations in commercial wines?

Spontaneous Fermentation What are we trying to achieve with a spontaneous fermentation? Longer presence of non-saccharomyces in early and mid stages of fermentation High diversity of indigenous strains of S. cerevisiae Higher complexity in aroma and flavour Do we achieve any of these when conducting spontaneous fermentations?

Spontaneous Fermentations Non-Saccharomyces Saccharomyces cerevisiae -Hanseniaspora/Kloeckera -Pichia -Metschnikowia Indigenous S. cerevisisae -Candida spp. Commercial S. cerevisisae

Species Identification Sample Spread Plate PCR amplification D1/D2 region Species Identification Pure Culture DNA extraction

Strain Identification 24 Sample Spread Plate PCR amplification Published Data Bases Yeast Strain Identification 8 Microsatellite loci Streak DNA extraction

Inoculated Fermentation Inoculum persistence from five different vintages in Pinot noir An inoculation is deemed successful when the inoculum is >80% relative abundance at the end of fermentation Year% No.%of%Wineries% Sampled% Tanks/Total%Tanks% with%>80%%inoculum% Percentage%of% Tanks%with% >80%%Inoculum% 2009$ 1$ 0/3$ 0$ 2010$ 3$ 3/9$ 33$ 2011$ 1$ 2/3$ 67$ 2012$ 3$ 5/8$ 63$ 2013$ 4$ 9/12$ 75$ $

CEDAR CREEK WINERY 2010 PINOT NOIR (JESSICA LANGE) END STAGE Relative Abundance/ 16 isolates 16 14 12 10 8 6 4 2 0 Tank 1 (L2TD) Cedar Creek: 'END-Stage' Population Dynamics in Guided & Spontaneous Fermentation Tanks Tank 2 (AMH) Tank 3 (RC212) Spontaneous Lallemand EC118 S. cerevisiae- Unknown 3 Zymaflore FX10 S. cerevisiae- Unknown 4 Lallemand Rhone2056 Zymaflore X5 Lallemand D47 RC212 D254 Non-Saccharomyces sp. (Lalvin EC1118)

CEDAR CREEK WINERY 2010 PINOT NOIR (JESSICA LANGE) END STAGE Cedar Creek: 'END-Stage' Population Dynamics in Guided & Spontaneous Fermentation Tanks Relative Abundance/ 16 isolates 16 14 12 10 8 6 4 2 0 Tank 1 (L2TD) Tank 2 (AMH) Tank 3 (RC212) Spontaneous Lallemand EC118 S. cerevisiae- Unknown 3 Zymaflore FX10 S. cerevisiae- Unknown 4 Lallemand Rhone2056 Zymaflore X5 Lallemand D47 RC212 D254 Non-Saccharomyces sp. (Lalvin EC1118) L2TD E-AMH RC212 L7213

CEDAR CREEK WINERY 2010 PINOT NOIR (JESSICA LANGE) END STAGE Relative Abundance/ 16 isolates 16 14 12 10 8 6 4 2 0 Tank 1 (L2TD) Cedar Creek: 'END-Stage' Population Dynamics in Guided & Spontaneous Fermentation Tanks * * * * Tank 2 (AMH) Tank 3 (RC212) Spontaneous Lallemand EC118 S. cerevisiae- Unknown 3 Zymaflore FX10 S. cerevisiae- Unknown 4 Lallemand Rhone2056 Zymaflore X5 Lallemand D47 RC212 D254 Non-Saccharomyces sp. (Lalvin EC1118) Zymaflore X5 RC212 L2TD E-AMH RC212 L7213 *Lalvin ICV D254 = Fermol Premier Cru

CEDAR CREEK WINERY 2010 PINOT NOIR (JESSICA LANGE) END STAGE Relative Abundance/ 16 isolates 16 14 12 10 8 6 4 2 0 * Tank 1 (L2TD) Cedar Creek: 'END-Stage' Population Dynamics in Guided & Spontaneous Fermentation Tanks * Tank 2 (AMH) * Tank 3 (RC212) * Spontaneous Lallemand EC118 S. cerevisiae- Unknown 3 Zymaflore FX10 S. cerevisiae- Unknown 4 Lallemand Rhone2056 Zymaflore X5 Lallemand D47 RC212 D254 Non-Saccharomyces sp. (Lalvin EC1118) L2TD E-AMH RC212 L7213 *ICV-D254 and Rhone 2056

Quails Gate Winery 2012 Pinot Noir (Erin Faasse) Percent Relative Abundance 1.00 0.75 0.50 0.25 0.00 CS (23.8 Brix) ER (22.79 Brix) M (15 Brix) F (0.6 Brix) Inoculated RC 212

Quails Gate Winery 2012 Pinot Noir (Erin Faasse) Percent Relative Abundance 1.00 0.75 0.50 0.25 0.00 CS (23.8 Brix) ER (22.79 Brix) M (15 Brix) F (0.6 Brix) Inoculated RC 212 Hanseniaspora uvarum

Quails Gate Winery 2012 Pinot Noir (Erin Faasse) Percent Relative Abundance 1.00 0.75 0.50 0.25 0.00 CS (23.8 Brix) ER (22.79 Brix) M (15 Brix) F (0.6 Brix) Inoculated RC 212

Quails Gate Winery 2012 Pinot Noir (Erin Faasse) Percent Relative Abundance 1.00 0.75 0.50 0.25 0.00 CS (23.8 Brix) ER (22.79 Brix) M (15 Brix) F (0.6 Brix) Inoculated RC 212 1.00 0.75 0.50 0.25 0.00 CS (24.24 Brix) ER (20.1 Brix) M (11.5 Brix) F (3.2 Brix) Spontaneous Fermentation RC212 Lalvin CY3079 Fermol Arome Plus ICV D-254

Spontaneous Fermentations At Four Wineries in 2013 (Scholl et al. 2016) Pinot Noir Chardonnay

Summary You do not know what is fermenting your wine unless you identify the organisms Inoculation does not guarantee a dominant implantation of the inoculum Over 5 years, 30% of the inoculated fermentations have <80% implantation Spontaneous fermentations usually contain a diversity of commercial S. cerevisiae strains Indigenous S. cerevisiae in spontaneous fermentations are rare Thus, wild, indigenous, and native are terms that do not accurately describe spontaneous fermentations

Answer To Questions Do we achieve what we want when conducting inoculated fermentations in commercial wines? Do we achieve what we want when conducting spontaneous fermentations in commercial wines?

Recommendations To know what yeasts are fermenting your must To view commercial yeast diversity as a good thing To start thinking about inoculating with a diversity of commercial and indigenous yeasts

Acknowledgements Financial Support BC Wine and Grape Council, NSERC Engage and Collaborative Research Development grants Quails Gate Estate Winery UBC Okanagan Biology Department Winery Assistance Grant Stanley from 50th Parallel Winery, Darryl Brooker from Cedar Creek Estate Winery/Mission Hill Family Estate Winery, David Patterson from Tantalus Vineyards and Niki Callaway and David Ledderhoff from Quails Gate Estate Winery Durall Lab Barb Hall, Amy Kluftinger, Jessica Lange, Frida Gustafsson, Laura Feeny, Marissa Neuner, Sarah Smith, Sydney Morgan, Chrystal Scholl, Mansak Tantikachornkiat, Erin Faasse, Morgan Stone, Natasha Benson, Michelle Stephenson.

Sydney Morgan Ph.D. Student, University of British Columbia

Effects of initial sulfur dioxide addition on wine yeast strains in spontaneous fermentations Sydney Morgan Supervisor: Dr. Daniel Durall University of British Columbia

SO 2 in winemaking SO 2 added to wine must before alcoholic fermentation for two main reasons: http://www.nytimes.com/2012/10/10/dini ng/bo nny-doon -vineya rds-la beling -policy-d raws-little -att ention. html?pagewa nted =all

SO 2 in winemaking SO 2 added to wine must before alcoholic fermentation for two main reasons: Antioxidant Prevent oxidation and browning reactions http://www.nytimes.com/2012/10/10/dini ng/bo nny-doon -vineya rds-la beling -policy-d raws-little -att ention. html?pagewa nted =all

SO 2 in winemaking SO 2 added to wine must before alcoholic fermentation for two main reasons: Antimicrobial agent Remove potential spoilage organisms (vineyard) http://www.nytimes.com/2012/10/10/dini ng/bo nny-doon -vineya rds-la beling -policy-d raws-little -att ention. html?pagewa nted =all

Why study SO 2 in wine? Health concerns Consumer demand for more natural food products SO 2 additions disallowed in organic wines http://www.bestwesternkelownahotel.com/blog/okanagan-attractions/kelowna-hotels-giv e-inside-scoop-on-golf-wine.html

Research objective Investigate effects of initial SO 2 addition on S. cerevisiae strains in spontaneous fermentations at a commercial winery http://www.fsrmagazine.com/wine/pinot- grigio -hits- right- notes http://www.vancouversun.com/life/cedarcr eek+s ale+m andl+ good +de al/945 4939/s tory. html

Experimental Design 0ppm SO 2 20ppm SO 2 40ppm SO 2

Strain composition

Commercial strains

Dominant strains

Dominant Strains

Sensory Analysis How do these treatments affect the sensory profiles of these wines?

Sensory Analysis Wines Reps (R Judges (J) (W) ) J x W J x R W x R MSE Citrus Aroma 33.65*** 2.92** 8.63** 1.22 3.79** 1.65 93.50 Pome Fruit Aroma 53.53*** 1.18 0.57 1.60* 3.99*** 0.62 114.7 Tropical Fruit Aroma 11.73*** 0.99 0.01 0.83 2.41* 0.73 246.3 Spice Aroma 44.20*** 1.18 1.41 0.76 5.58*** 0.61 97.15 Vanilla Aroma 18.78*** 1.32 10.53** 0.76 2.48* 0.99 220.8 Toasty/Smoky Aroma 27.99*** 2.35* 0.67 1.27 8.04*** 0.87 108.5 Unprocessed Wood Aroma 32.05*** 0.64 4.75* 0.89 3.00** 0.28 218.2 Citrus Flavour 35.75*** 0.29 0.49 1.31 6.37*** 1.03 46.85 Pome Fruit Flavour 96.56*** 1.45 1.58 0.87 3.64** 0.65 69.31 Tropical Fruit Flavour 72.28*** 2.09* 0.84 1.10 15.38*** 0.90 65.66 Spice Flavour 94.40*** 0.71 3.53 0.92 4.49*** 1.82 46.80 Vanilla Flavour 78.52*** 1.00 20.07*** 1.35 1.93 1.54 58.84 Toasty/Smoky Flavour 88.82*** 0.25 4.44* 1.24 5.28*** 0.61 53.67 Unprocessed Wood Flavour 40.81*** 0.79 29.38*** 1.02 2.90** 0.46 74.8 Acidity 73.85*** 0.59 21.32*** 1.13 2.55* 0.46 74.60 Astringency 89.25*** 0.28 0.19 0.53 2.98** 1.17 104.2 Body 95.40*** 0.63 2.31 0.87 9.40*** 1.39 36.46 Length of Aftertaste 35.43*** 0.96 2.57 1.09 3.92*** 1.16 47.74

Sensory Analysis Judges (J) Wines (W) Reps (R) J x W J x R W x R MSE Citrus Aroma 33.65*** 2.92** 8.63** 1.22 3.79** 1.65 93.50 Pome Fruit Aroma 53.53*** 1.18 0.57 1.60* 3.99*** 0.62 114.7 Tropical Fruit Aroma 11.73*** 0.99 0.01 0.83 2.41* 0.73 246.3 Spice Aroma 44.20*** 1.18 1.41 0.76 5.58*** 0.61 97.15 Vanilla Aroma 18.78*** 1.32 10.53** 0.76 2.48* 0.99 220.8 Toasty/Smoky Aroma 27.99*** 2.35* 0.67 1.27 8.04*** 0.87 108.5 Unprocessed Wood Aroma 32.05*** 0.64 4.75* 0.89 3.00** 0.28 218.2 Citrus Flavour 35.75*** 0.29 0.49 1.31 6.37*** 1.03 46.85 Pome Fruit Flavour 96.56*** 1.45 1.58 0.87 3.64** 0.65 69.31 Tropical Fruit Flavour 72.28*** 2.09* 0.84 1.10 15.38*** 0.90 65.66 Spice Flavour 94.40*** 0.71 3.53 0.92 4.49*** 1.82 46.80 Vanilla Flavour 78.52*** 1.00 20.07*** 1.35 1.93 1.54 58.84 Toasty/Smoky Flavour 88.82*** 0.25 4.44* 1.24 5.28*** 0.61 53.67 Unprocessed Wood Flavour 40.81*** 0.79 29.38*** 1.02 2.90** 0.46 74.8 Acidity 73.85*** 0.59 21.32*** 1.13 2.55* 0.46 74.60 Astringency 89.25*** 0.28 0.19 0.53 2.98** 1.17 104.2 Body 95.40*** 0.63 2.31 0.87 9.40*** 1.39 36.46 Length of Aftertaste 35.43*** 0.96 2.57 1.09 3.92*** 1.16 47.74

Sensory Analysis

Conclusions SO 2 addition at crush resulted in significantly altered S. cerevisiae strain compositions

Conclusions Previously used commercial strains dominate spontaneous fermentations regardless of SO 2 addition level

Conclusions SO 2 addition level at crush resulted in sensory differences in the final wines http://www.adglobalgroup.com/wp-content/uploads/2015/08/wine-tasting2.jpg

Acknowledgements Winemakers at Cedar Creek Estate Winery Darryl Brooker, Taylor Whelan All members of the Durall lab at UBC Natasha Benson, Chrystal Scholl, Morgan Stone, Marissa Neuner, Ben Tantikachornkiat