本研究挑選了四個台灣中部之商業栽培葡萄園進行葡萄成熟過程中酵母菌數與菌相變化相關研究。本研究利用平板計數法計算樣品中之酵母菌數，同時經CHROMagar與隨機擴增多型性DNA (Random amplified polymorphism DNA, RAPD)進行分群後，以大單元核糖體DNA D1D2區序列，並分析各區葡萄園中各樣品菌相變化情形。結果顯示：各區域各時期菌數變化非常相似，各品種間會有明顯差異。另菌數變化與套袋與否不具相關性，惟菌數會隨葡萄成熟度增加而上升，未成熟果實表面菌數約<1~3.33 Log (CFU/g)，至成熟期後菌數則可達2.39~5.84 Log (CFU/g)，而葉面菌數則明顯高於果實，菌數約為<1~6.24 Log (CFU/g)。化學特性分析結果顯示：糖含量與pH會隨著葡萄成熟而上升，並與菌數成正比，酸度則隨葡萄成熟度增加而降低，且其菌數隨酸度降低而增加。各樣區葡萄園主要優勢菌種皆為Sporidiobolus pararoseus，其餘較常見則有Cryptococcus sp. (09)、Pseudozyma aphidis與Pseudozyma antarctica等菌種，各地區菌相變化有所不同，以新社地區未套袋巨峰葡萄為例，著果期與轉色期以Sporidiobolus pararoseus為主要優勢菌株，至成熟期優勢菌種變為Jaminaea angkoriensis，到了收穫期主要優勢菌株則為Cryptococcus sp. (09)與Pseudozyma aphidis等菌株。本研究所有樣品中除新社地區成熟期巨峰葡萄分離出3株Saccharomyces cerevisiae外，其餘區域皆未發現該菌種，此結果與前人研究結論相符，亦即Saccharomyces cerevisiae並非葡萄表面優勢菌種。除上述描述菌種外，其他菌種共有20個屬48個種，較常見菌種包括Sporidiobolus pararoseus, Pseudozyma aphidis與Aureobasidiumpullulans var. melanogenum等菌株。

In this study, we investigated the dynamics of yeast counts and flora changes on fruits and leaves during fruit ripening. The total counts of yeasts on grapes and leaves sampled from four commercial vineyards were evaluated by plate count method. Then approximately fifty colonies were picked randomly from plates, and grouped by their morphology and color revealed on Chromagar. All representive strains in each groups by above method were clustered by randomly amplified polymorphism DNA (RAPD) with primer (GTG5). At the last, these representive strains were identified by the sequence comparison of D1/D2 domain of the 26S ribosomal RNA gene. The results showed that total yeast counts was increased with the ripened grapes. but variable in different grape varieties. Total yeast count of grapes in maturation stage ranged from <1 to 3.33 Log (CFU/g) and 2.39 to 5.84 Log (CFU/g). Also, the total yeast counts on leaves <1 to 6.24 Log (CFU/g) are higher than those on grapes <1 to 3.33 Log (CFU/g). Sugar concentration and pH value in grapes were raised during fruit ripening, but acidity was decreased at the same period. Total yeast count was increased with the decrease of acidity. The dominant yeast species of four vineyards were Sporidiobolus pararoseus, Cryptococcus sp.(09), Pseudozyma aphidis, and Pseudozyma antarctica. The yeast flora were different in different vineyard. In Xinshe, for example, Sporidiobolus pararoseus was the dominant species of unpackage grapes in fruiting and veraison stage, but Jaminaea angkoriensis and Cryptococcus sp.(09) were the dominant ones in maturation and harvest stage, Moreover, we isolated only three Saccharomyces cerevisiae strains from one of the four vineyards, which indicated that Saccharomyces cerevisiae is difficult to be isolated from grape surface,which comcident with studies by previous author. The total isolates in this study can be identified into 20 species and 48 strains, and Sporidiobolus pararoseus, Pseudozyma aphidis, and Aureobasidium pullulans var. melanogenum were the common species.

陸、參考文獻
白秋菊、白旭凱、謝廉一。2013。102年5月主要農作物生產預測。農政與農情。 251:120~126
連忠勇、張致盛。2004。葡萄產業現況與輔導策略。臺中區農業改良場特刊。 67:1-8
劉興隆、趙佳鴻、沈原民、吳世偉。2010。亞磷酸對葡萄主要病害之影響。臺中區農業改良場特刊。107:123-133
林月金。2007。臺灣鮮食葡萄之產銷研究。農業經營管理專輯特刊。76:79-96。
林嘉興。1988a。改善目前的葡萄整枝與修剪技術。葡萄生產技術14:173-188。
林信山、林嘉興。1978。乙撐氯醇在葡萄栽培上之利用。台灣農業 14(4):83～89。
林嘉興。1988b。葡萄休眠與催芽技術。葡萄生產技術14:189-196。
林嘉興。2004。葡萄產業沿革與栽培技術之發展。葡萄栽培技術研討會專集9〜22。
林玉靜。2012。土壤及有機物質中之木醣發酵菌分離、鑑定及酒精產量分析。新竹教育大學生命科學系碩士論文。
黃杉坁。1992。非農藥防治作物病蟲[作物套袋栽培]。台南區農業改良場技術專刊植保-6 59:1-4。
邱禮弘、張致盛。2010。巨峰葡萄合理化施肥技術。行政院農業委員會臺中區農業改良場特刊100:213-218。
謝智雯。2009。台灣南部地區土壤酵母菌種類分佈與調查。新竹教育大學生命科學系碩士論文。
張致盛。2007。台灣栽培葡萄品種之特性。台灣區農業專刊:1-4。
張致盛、張林仁、林嘉興。2004。台灣葡萄生產產期調節技術。葡萄栽培技術研討會專集37〜53。
鍾伊婷。2004。台灣食用及釀酒葡萄果實生長期及葡萄酒發酵期間酵母菌菌相變化調查。東吳大學微生物學系碩士論文。
楊尚勳。2006。葡萄優質化農藥減量策略。農政與農情165:88。
楊耀祥。1995。葡萄。台灣農家要覽。農作篇(二):183-190。
Achstetter, T., and Wolf, D.H. (1985). Proteinases, proteolysis and biological control in the yeast Saccharomyces cerevisiae. Yeast. 1: 139-157.
Buck, J.W., and Burpee, L.L. (2002). The effects of fungicides on the phylloplane yeast populations of creeping bentgrass. Canadian Journal of Microbiology. 48: 522–529.
Buglione, M., and Lozano, J. (2002). Nonenzymatic browning and chemical changes during grape juice storage. Journal of Food Science. 67:1538-1543.
Combina, M., Mercado, L., Borgo, P., Elia, A., Jofre´, V., Ganga, A., Martinez, C. (2005). Yeasts associated to Malbec grape berries from Mendoza, Argentina. Journal of Applied Microbiology. 98:1055–1061.
Cˇ adezˇ, N., Zupan, J., Raspor, P. (2010). The effect of fungicides on yeast communities associated with grape berries. FEMS Yeast Research. 10:619–630.
Chi, Z., Wang, F., Yue, L., Liu, G., Zhang, T. (2009). Bioproducts from Aureobasidium pullulans, a biotechnologically important yeast. Applied Microbiology and Biotechnology. 82:793–804.
Chavan, P., Mane S., Kulkarni, P., Shaikh, S., Ghormade, V., Nerkar, D.P., Shouche, Y., Deshpande, M.V. (2009). Natural yeast flora of different varieties of grapes used for wine making in India. Food Microbiology. 26:801–808.
Davenport, R.R. (1974). Microecology of yeasts and yeast-like organisms associated with an English vineyard. Vitis. 13:123-130.
Dimakopoulou, M., Tjamos, S.E., Antoniou, P.P., Pietri, A., Battilani, P., Avramidis, N., Markakis, E.A., Tjamos, E.C. (2008). Phyllosphere grapevine yeast Aureobasidium pullulans reduces Aspergillus carbonarius (sour rot) incidence in wine-producing vineyards in Greece. Biological Control. 46:158–165.
Erez, A. and Zur, A. (1981). Breaking the rest of apple buds by narrowdistillation range oil and dinitro-o-cresol. Scientia Horticulturae. 14:47-54.
Fleet, G.H. (2003). Yeast interactions and wine flavor. International Journal of Food Microbiology. 86:11–22.
Fleet, G.H., Prakitchaiwattana, C., Beh, A.L., Heard, G. (2002). The yeast ecology of wine grapes. Biodiversity and Biotechnology of Wine Yeasts. 1-17
Guerra, E., Sordi, G., Mannazzu, I., Clementi, F., Fatichenti, F. (1999). Occurrence of wine yeasts on grapes subjected to different pesticide treatments. Italian Journal of Food Science. 11: 221–230.
Gildemacher, P.R., Heijne, B., Houbraken, J., Vromans, T., Hoekstra, E.S., Boekhout, T. (2004). Can phyllosphere yeasts explain the effect of scab fungicides on russeting of Elstar apples? European Journal of Plant Pathology. 110: 929–937.
Gholamnejad, J., Etebarian, H.R., Sahebani, N. (2010). Biological control of apple blue mold with Candida membranifaciens and Rhodotorula mucilaginosa. African Journal of Food Science. 4:1-7.
Hagler A.N. (2006) Yeasts as indicators of environmental quality. Biodiversity and Ecophysiology of Yeasts. 21:519–536.
Huang, R., Che, H.J., Zhang, J., Yang, L., Jiang, D.H., Li, G.Q. (2012). Evaluation of Sporidiobolus pararoseus strain YCXT3 as biocontrol agent of Botrytis cinerea on post-harvest strawberry fruits. Biological Control. 62:53–63.
Ippolito, A., Ghaouth, A. EI., Wilson C.L., Wisniewski, M. (2000). Control of post- harvest decay of apple fruit by Aureobasidium pullulans and induction of defense responses. Postharvest Biology and Technology. 19:265–272.
Jolly, N.P., Augustyn, O.P.H., Pretorius I.S. (2003). The occurrence of non-Saccharomyces cerevisiae yeast species over three vintages in four vineyards and grape musts from four production regions of the western cape, South Africa. South African Journal for Enology & Viticulture. 24: 35-42.
Kurtzman, C.P., and Robnett, C.J. (1998). Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek. 73: 331–371.
Kulakovskaya, T.V., Shashkov, A.S., Kulakovskaya, E.V., Golubev, W.I. (2004). Characterization of an antifungal glycolipid secreted by the yeast Sympodiomycopsis paphiopedili. FEMS Yeast Research. 5:247–252.
Longo, E., Cansado, J., Agrelo, D., Villa, T.G. (1991). Effect of climatic conditions on yeast diversity in grape musts from Northwest Spain. American Journal of Enology and Viticulture. 42:141-144.
Loureiro, V. and Ferreira, M.M. (2003). Spoilage yeasts in the wine industry. International Journal of Food Microbiology. 86:23–50.
Martini, A., Federici, F., Rosini, G. (1980). A new approach to the study of yeast ecology of natural substrates. Canadian Journal of Microbiology. 26:856-859.
Morita, T., Konishi, M., Fukuoka, T., Imura, T., Kitamoto, D. (2007). Physiological differences in the formation of the glycolipid biosurfactants, mannosylerythritol lipids, between Pseudozyma antarctica and Pseudozyma aphidis. Applied Microbiology and Biotechnology. 74:307–315.
Morita, T., Konishi, M., Fukuoka, T., Imura, T., Kitamoto, D. (2007). Microbial conversion of glycerol into glycolipid biosurfactants, mannosylerythritol lipids, by a basidiomycete yeast, Pseudozyma antarctica JCM 10317T. Journal of bioscience and bioengineering. 104:78–81.
Morita, T., Fukuoka, T., Imura, T., Kitamoto, D. (2013). Accumulation of cellobiose lipids under nitrogen-limiting onditions by two ustilaginomycetous yeasts, Pseudozyma aphidis and Pseudozyma hubeiensis. FEMS Yeast Research. 13:44–49.
Mounir, R., Durieux, A., Bodo, E., Allard, C., Simon, J.P., Achbani, E.H., Jaafari, S.EI., Douira, A., Jijakli, M.H. (2007). Production, formulation and antagonistic activity of the biocontrol like-yeast Aureobasidium pullulans against Penicillium expansum. Biotechnology Letters. 29:553–559.
Renouf, V., Claisse, O., Funel, A.L. (2005). Understanding the microbial ecosystem on the grape berry surface through numeration and identification of yeast and bacteria. Australian Journal of Grape and Wine Research. 11:316-327.
Rementeria, A., Rodriguez, J.A., Cadaval, A., Amenabar, R., Muguruza, J.R., Hernando F.L., Sevilla, M.J. (2003). Yeast associated with spontaneous fermentations of white wines from the ‘‘Txakoli de Bizkaia’’ region (Basque Country, North Spain). International Journal of Food Microbiology. 86 :201– 207.
Rosini, G., Federici, F., Martini A. (1982). Yeast flora of grape berries during ripening. Microbial Ecology. 8:83-89.
Radler, F., Herzberger, S., Schonig, I., Schwarz, P. (1993). Investigation of a killer strain of Zygosaccharomyces bailii. Journal of General Microbiology. 139:495-500.
Sabat, J., Cano, J., Zarzoso, B.E., Guillamón, J.M. (2002). Isolation and identification of yeasts associated with vineyardand winery by RFLP analysis of ribosomal genes and mitochondrial DNA. Microbiological Research. 157: 267–274.
Tolaini, V., Reverberi, M., Fanelli, C., Fabbri, A.A., Fiore, A.D., De Rossi, P., Ricelli A. (2010). Lentinula edodes enhances the biocontrol activity of Cryptococcus laurentii against Penicillium expansum contamination and patulin production in apple fruits. International Journal of Food Microbiology. 138:243–249.
Walczak, E., Czaplin’ska, A., Barszczewski, W., Wilgosz, M., Wojtatowicz, M., Robak, M. (2007). RAPD with microsatellite as a tool for differentiation of Candida genus yeasts isolated in brewing. Food Microbiology. 24:305–312.
Walter, M., Frampton, C.M., Boyd-Wilson, S.H., Harris-Virgin, P., Waipara, N.W. (2007). Agrichemical impact on growth and survival of non-target apple phyllosphere microorganisms. Canadian Journal of Microbiology. 53:45–55.
Xu, B., Zhang, H., Chen, K., Xu, Q., Yao, Y., Gao, H. (2013). Biocontrol of postharvest Rhizopus decay of peaches with Pichia caribbica. Current Microbiology. 67:255–261.
Zhang, S., Schisler, D.A., Boehm, M.J., Slininger, P.J. (2007). Utilization of chemical inducers of resistance and Cryptococcus flavescens OH 182.9 to reduce Fusarium head blight under greenhouse conditions. Biological Control. 42:308–315.
Zhang, H.Y., Zheng, X.D., Xi, Y.F. (2005). Biological control of postharvest blue mold of oranges by Cryptococcus laurentii (Kufferath) Skinner. BioControl. 50: 331–342.