Study On The Diversity Of Non-Saccharomyces Yeast Of Cabernet Saovignon From Differfent Sub-regions At The East Foothillof Helan Mountain

At present,the research on Saccharomyces saccharomyces savannaii is not limited to Saccharomyces cerevisiae,and non-Saccharomyces cerevisiae has gradually become the main research object at home and abroad.Taking advantage of the good geographical resources in the eastern foothills of Helan Mountain,isolating the local representative non-Saccharomyces cerevisiae is one of the current local research priorities.The research object of this study is mainly based on Cabernet Sauvignon grapes from five sub-producing regions in the eastern foothills of Helan Mountain as raw materials,and the non-Saccharomyces cerevisiae strains isolated during natural fermentation are identified by WRN culture medium morphology and 26S r DNA D1/D2 region sequence analysis,and the non-Saccharomyces cerevisiae strains are analyzed by Tandem repeat-tRNA(TRtRNA)fingerprinting method for interspecific and intraspecific molecular fingerprinting,and the results of the study are as follows.(1)A total of 524 non-saccharomyces cerevisiae strains were isolated from five wine producing regions at the eastern foot of Helan Mountain,and identified by WLN medium morphological classification and 26S R DNA D1/D2 region sequence analysis,which were classified into 19 species belonging to 10 genera:Hanseniaspora uvarum,Metschnikowia Pimensis,Metschnikowia pulcherrima,Cryptococcus sp.,Cryptococcus albidus,Cryptococcus flavescens,Cystofilobasidium ferigula,Cryptococcus terrestris,Rhodotorula glutinis,Naganishia albida,Candida Zemplinina,Pichia Kluyveri,Pichia Kudriavzevii,Rhodotorula Graminis,Aureobasidium pullulans,Filobasidium Magnum,Filobasidium sp,Cryptococcus magnus,Filobasidium elegans.(2)There are 7 genera and 11 species of yeast in Shizuishan producing area,accounting for 57.89%of the total yeast species.There are 5 genera and 10 species of yeast in Qingtongxia production area,accounting for 52.63%of the total yeast species;Yinchuan production area has 8 genera and 8 species,accounting for 42.1 1%of the total yeast species;Yeast in Hongsipu production area There are 3 genera and 4 species of yeast,accounting for 21.05%of the total yeast species;there are 3 genera and 3 species of yeast in Yuquanying production area,accounting for 15.79%of the total yeast species.There are 2 genera and 2 species of yeast unique to Yinchuan production area,namely:R graminis and A.pullulans,and 4 genera and 4 species of yeast unique to Shizuishan production area,namely:C.ferigula,C.terrestris,R.glutinis,N.albida,there are 2 genera and 3 species of yeasts unique to Qingtongxia production area,namely:Filobasidium sp.,C.magnus and F.elegans,and only 1 yeast species unique to Hongsipu production area Genus 1 species,namely:P.kudriavzevii.The yeast strains of H.uvarum and M.pulcherrima were isolated from the five production areas of Yinchuan,Shizuishan,Qingtongxia,Yuquanying and Hongsibao,of which H.uvarum was the dominant strain in the five sub-producing areas.Among them,18 new yeast strains were isolated from the naturally fermented Cabernet Sauvignon wine this time,accounting for 3.44%of the total strains isolated.(3)The combination of traditional methods and molecular biological methods can effectively realize the interspecific and intraspecific studies of non-saccharomyces cerevisiae.Primer pairs(4)TtRNASC and 5CAG divided all non-saccharomyces cerevisiae strains into 31 categories,primer pairs TtRNASC and ISSR-MB all non-saccharomyces cerevisiae strains into 37 categories,n.albida did not show fingerprints.Cryptococcus and other yeast had the same differentiation effect between the two pairs of microsatellite primers.The effect of H.uvarum and other yeast on the intraspecies differentiation was higher in primer pairs than in primer pairs.19 species of non-saccharomyces cerevisiae belonging to 10 genera showed certain interspecific and intraspecific differences in the fingerprints of two pairs of microsatellite primers,indicating that H.uvarum,C.zemplinina and Cryptococcus strains isolated from the natural fermentation of Cabernet sauvignon grapes at the eastern piedmont of Helan Mountain were rich in genetic diversity.The genetic diversity of M.pulcherrima,Cystofilobasidium,Pichia and other yeast strains was relatively single.