Better Whiskey Through Science! Leveraging a public corn breeding program to improve regional whiskey flavors and create new industries

Rob Arnold, MSc Seth C. Murray, PhD Head Distiller Associate Professor and Eugene Butler Endowed Chair Firestone & Robertson Distilling Co. Dept. of Soil & Crop Sciences PhD student, Texas A&M University Texas A&M University @TXDistiller @DrSethMurray What is Whiskey?

https://www.winebags.com/The-Compendium-of-Alcohol-Ingredients-and-Processes-s/2054.htm Whiskey: generic. Distilled alcohol made with grain and aged in a wood Most U.S. whiskeys use a lot of corn : US Law: 51+% corn, distilled <80% alcohol, barreled at < 62.5% alcohol, new charred oak barrel Tennessee whiskey: Usually filtered through charcoal Texas whiskey: The best whiskey! : 51+% rye (grain) Scotch : Usually malted barley, distilled 2 times in Scotland : Usually malted barley, distilled 3 times and aged 3 years in Ireland Most corn (, Zea mays L.) is not Corn - $51.5 billion, 89 million acres, 404 million tons Sweet corn - $860 million, 495,000 acres, 4 million tons Plant breeding and agronomy have increased yields

Brummer, E.C., W.T. Barber, S. Collier, T.S. Cox, R. Johnson, S.C. Murray*, R.T. Olsen, R.C. Pratt, and A.M. Thro. 2011. Plant breeding for harmony between agriculture and the environment. Frontiers of Ecology and the Environment 9:561–568.

Data from USDA- NASS A need to feed the growing world population Nobel Laurate Dr. 10 billion people Norman Borlaug by 2050 “Father of the Green Revolution” Former Texas A&M Faculty Member

http://www.worldfoodprize.org/ http://esa.un.org/unpd/wpp/Graphs/Probabilistic/POP/TOT/ 170 U.S. corn yield in bushels per cre from 1866 to 2016 Period of industry 150 “Synthetic fertilizer” Haber-Bosch process consolidation <1908 1996 130 Public breeding Industry sales “Hybrid vigor” 2012 110 1930 - 1975 Drought Darwin Shull and East Genomic selection 1876 1908/9 developed in 2007 90 Yield bu/A Yield 70 H. Wallace first hybrid seed sale First genetic maps 1986 1924 50 ‘B73’ developed 1973 Open Pollinated DNA major discoveries 0.008 Bu/A/yr 30 Watson & Crick Sanger 1953 10 1860 1880 1900 1920 1940 1960 1980 2000 2020 Year Now a $12 billion dollar private corn seed industry Improvements in addition to yield are needed Reduced input use Ecosystem service provision Flavor Human health & nutrition Climate change resilience Carbon

http://www.nnvl.noaa.gov/ Corn Wild species X Zea Wild species hybrid

Zea mays (Corn) Commercial Hybrid New lines selected from crosses Native (“natural”) genetic diversity in maize

The International Maize and Wheat Improvement Center (CIMMYT) - https://maize.org/maize-from-mexico-to-the-world/ How do we breed corn? – 30,000ft. view Tassels: male pollen 1. Develop inbred lines A. Cross best parents B. Self 2. Make hybrid seed with Shoot another inbred bags 3. Yield trials and measure other traits Ear: 4. Only advance the best female silks

5. Extensive testing Tassel bags 6. Sell seed to farmers Pace of change is slow in plant breeding

7 years

Nokia 2330 http://www.mymobiles.com/ Samsung S6 http://www.mymobiles.com/

1 new variety Modern genomics and phenomics tools speed breeding p1; Red cob Blue r1 c1 grain y1; Yellow grain

http://www.illumina.com/ Chromosome

Dr. Sorin Popescu Yet, a lot of manual labor is still required by dedicated students and staff Problems facing Texas (& Southern) corn farmers 1. Very low yield & not increasing like midwest • Industry consolidation to the Corn Belt • Texas alone has $1.5 billion in corn value

2. Heat and drought stress * At the forefront of climate change

3. Aflatoxin

4. Markets Texas is one of last states with a public corn breeding program (and has two)! Novel corn for whiskey in Texas A&M Corn Breeding Program Evaluate ~7,000 varieties for testing /year

● Variety 1 ● Variety 5 ● Variety 2 ● Variety 6 ● Variety 3 ● Variety 7 ● Variety 4 ● Variety 8 Can different corn’s make whiskey taste different? better? No scientific publications to date on corn whiskey flavor! Rob Arnold, MSc - 3rd generation distiller - Selected his own yeast strain - Wants to develop a corn variety - Doing a distance PhD at Texas A&M Turning to science… Step 1: Develop a repeatable method to test small samples

Scientific microbatch (1L) new-make distilling Step 2: Develop new methods for measurement Fermentation Gas Sensory panels Good Bad & chromatography- Anise Sour Banana Woody • Alcohol yield olfactometry Caramel MustyDusty Chemicals Coffee Haylike • pH Corn Green contributing to DarkFruit Burnt Floral Buttery • Speed of odor and flavor Honey Vinegar fermentation Leather LacticAcid Malt Cardboard Mint Stale • Distillation proof Molasses Soapy Nutty Oily Pepper Rancid etc… Roast Fishy Smokey Butyric Vanilla Sulphur etc. etc. Step 3: Evaluate G, E, and G x E variation effects Does the hybrid variety grown matter? Genetics!

Do the environments grown matter? Environment!

Do hybrid varieties interact with the environment? Genetics x Environment interaction!

Not enough samples to look at management or microbes or … yet What have we found to date? • Evaluated ~30 Texas A&M varieties so far (PLOS ONE - in revision)

• Substantial impacts from G, E, and/or GxE ● Variety 1 ● Variety 5 ● Variety 2 ● Variety 6 ● Variety 3 ● Variety 7 • Alcohol yield: (G) variety = (E) environment > GxE ● Variety 4 ● Variety 8

• Flavor, aroma and congeners: each differed !!! terroir

• Different as new make spirit but differs more after aging

• Scaling up a proprietary Texas A&M hybrid

• Lots of popular press What have we found to date? Ethyl Octanoate correlated with good aromas (PLOS ONE - in revision)

v

2-Nonenal correlated with bad aromas

26 22 Public breeding programs distribution in the U.S.

Dr. Stephenhttp://css.wsu.edu S. Jones https://www.stonebarnscenter.org Wheat Breeder Dr. Michael R Mazourek Washington State University Vegetable breeder Cornell University

Dr. Christine Diepenbrock Plant Biology for Nutrition Shelton, A. C., & Tracy, W. F. (2017). Cultivar development in Dr. Stephen Kresovich UC Davis the US public sector. Crop Science, 57(4), 1823-1835. Clemson University US public plant breeders skew much older And less than half believe their position will be refilled after retirement

Shelton, A. C., & Tracy, W. F. (2017). Cultivar development in the US public sector. Crop Science, 57(4), 1823-1835. @DrSethMurray Breeding program funding Whiskey Murray group Steven Anderson research Jacob Pekar funding David Rooney Nathalia Cruzato* Colby Ratcliff Holly Lane Alper Adak Dr. Scott Wilde Firestone & Shakirah Nakasagga Robertson Many past students Distilling Co. and student workers – thank you! *Supported in-part by a USDA-NIFA-AFRI Award No. 2017-67013-26185, USDA- Monsanto future leaders NIFA Hatch funds, Texas A&M AgriLife Research, the Texas Corn fellowship Producers Board, the Iowa Corn Promotion Board and the Eugene Butler Endowed Chair in Biotechnology What are we tasting today?

Commercial TX whiskey Commercial Variety TAMU experimental made with Produced on John hybrids produced in commercial Sawyers Farm in Hill Burleson County, TX corn (for now) County, TX 2017 2017 aged four years 59% ABV 59% ABV in (119 proof) (119 proof) 40% ABV Both aged in for 1.5 years (80 proof) in matched oak barrels