Ayumi Abe, Tomoyuki Sato, and Teruo Sone*
*Research Faculty of Agriculture, Hokkaido University, Kita 9 Nishi 9, Kita-ku, Sapporo Hokkaido 060-8589, Japan (sonet@agr.hokudai.ac.jp)

In recent years, the number of wineries in Hokkaido, Japan, has increased significantly. Microorganisms are an important factor responsible for terroir. The purpose of this study was to collect wine-related microorganisms from various materials in Hokkaido and perform molecular identification of the isolates and their genetic and physiological characteristics. They can be used as starters in the future or as research materials. In this study, we focused on isolating yeasts and lactic acid bacteria such as Saccharomyces cerevisiae and Oenococcus oeni. We attempted to isolate them from various fruits, must under fermentation, and wine lees. For fruits with a lot of juice, such as grapes, the fruits were crushed in a plastic bag and allowed to ferment at room temperature. For strawberries, prunes, and other fruits with little juice, we added the fruit to commercial grape juice and waited for fermentation to occur at room temperature. Fermented samples were then spread on agar plates to form colonies. Colonies were isolated and identified by rDNA ITS or 16S rDNA. Strains isolated from the same sample and identified as S. cerevisiae and O. oeni were excluded from possible clones by SSR (Simple Sequence Repeat) analysis and RAPD (Random Amplified Polymorphic DNA) or SSR analysis, respectively, and were registered as AHU (Agriculture, Hokkaido University) collections. From 2016 to 2022, we collected 475 Saccharomyces strains and 143 O. oeni strains. Other LABs including Leuconostoc mesenteroides, Lactococcus lactis, Lactobacillus plantarum, and Lactobacillus brevis were isolated as well. We are currently analyzing the genetic and physiological characteristics of these collected strains.

Funding Support: Institution of Fermentation, Osaka/ Sapporo Bioscience Foundation