刺梨自然发酵过程中非酿酒酵母多样性分析

Biodiversity of non-Saccharomyces yeasts during natural fermentation of Rosa roxburghii

摘要:

[目的] 分析刺梨果实自然发酵过程中非酿酒酵母菌群特征，为筛选优质刺梨非酿酒酵母提供参考。[方法] 基于Illumina MiSeq高通量测序技术和WL营养琼脂鉴定培养基纯种分离技术，分析刺梨果实自然发酵1 d（F1）、3 d（F3）、5 d（F5）和15 d（F15）4个阶段及YPD培养基富集培养样本中非酿酒酵母种群组成和多样性。[结果] 高通量测序分析结果共获得182个OTUs（operational taxonomic units，OTUs），归属于81个属107个种；物种多样性分析结果表明，刺梨果实自然发酵前期，优势非酿酒酵母为汉逊酵母（Hanseniaspora sp.）和伯顿丝孢毕赤酵母（Hyphopichia burtonii），二者在样本F1中分别占42.59%和26.85%；随着自然发酵的不断进行，二者的比例逐渐降低，在第15天（F15），Hanseniaspora sp.和H.burtonii比例降低至7.73%和0.52%。相反，Pichia sporocuriosa和未培养的酵母，随着自然发酵不断进行所占比例逐渐增大，分别由F1中的0.23%和0.33%增至F15中的37.26%和32.62%。此外，采用WL营养琼脂鉴定培养基纯种分离和鉴定技术，从刺梨上分离到Hanseniaspora sp.、H.burtonii、克鲁维毕赤酵母（Pichia kluyveri）、P.sporocuriosa和异常威克汉姆酵母（Wickerhamomyces anomalus）5种类型的可培养非酿酒酵母。[结论] 刺梨果实上存在着丰富的非酿酒酵母菌资源，研究刺梨自然发酵过程中非酿酒酵母多样性，为酵母资源开发和利用奠定基础。

Abstract:

[Objective] The present study aims to reveal non-Saccharomyces yeasts biodiversity and theirs changes during natural fermentation of Rosa roxburghii fruit, so as to provide reference for the screening of high quality non-Saccharomyces yeasts.[Methods] The composition and biodiversity in non-Saccharomyces yeasts communities in four natural fermentation stages including 1 d (F1), 3 d (F3), 5 d (F5) and 15 d (F15) and the cultures from R. roxburghii in YPD medium were analyzed using high-throughput sequencing and WL (Wallerstein laboratory nutrient agar) identification medium.[Results] High-throughput sequencing results show that a total of 182 operational taxonomic units (OTUs) were detected and assigned to 81 species in 107 genera. Hanseniaspora sp. and Hyphopichia burtonii were predominant in R. roxburghii fruit, accounting for 42.59% and 26.85% in F1, respectively. With the continuous progress of natural fermentation, the content of Hanseniaspora sp. and H. burtonii decreased gradually. At the end of fermentation (F15), the proportion of Hanseniaspora sp. and H. burtonii decreased to 7.73% and 0.52%, respectively. In contrast, the proportion of Pichia sporocuriosa and unclassified_o_Saccharomycetales increased from 0.23%, 0.33% in F1 to 37.26%, 32.62% in F15, respectively. Moreover, Hanseniaspora_sp., H. burtonii, Pichia kluyveri, P. sporocuriosa and Wickerhamomyces anomalus were also isolated by pure culture approach.[Conclusion] There were abundant resources of non-Saccharomyces yeasts in R. roxburghii fruit. Study on the biodiversity of non-Saccharomyces yeasts during natural fermentation of R. roxburghii, it aims to lay a foundation for the exploitation and utilization of these yeast resources.