Development of Genetic Modification Tools for Hanseniaspora uvarum

被引:18
作者
Badura, Jennifer [1 ]
van Wyk, Niel [1 ,2 ]
Brezina, Silvia [1 ]
Pretorius, Isak S. [2 ]
Rauhut, Doris [1 ]
Wendland, Juergen [1 ]
von Wallbrunn, Christian [1 ]
机构
[1] Hsch Geisenheim Univ, Dept Microbiol & Biochem, Von Lade Str 1, D-65366 Geisenheim, Germany
[2] Macquarie Univ, Dept Mol Sci, ARC Ctr Excellence Synthet Biol, Sydney, NSW 2113, Australia
基金
澳大利亚研究理事会;
关键词
Hanseniaspora uvarum; transformation; genetic modification; ATF1; ethyl acetate; fermentation;
D O I
10.3390/ijms22041943
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Apiculate yeasts belonging to the genus Hanseniaspora are commonly isolated from viticultural settings and often dominate the initial stages of grape must fermentations. Although considered spoilage yeasts, they are now increasingly becoming the focus of research, with several whole-genome sequencing studies published in recent years. However, tools for their molecular genetic manipulation are still lacking. Here, we report the development of a tool for the genetic modification of Hanseniaspora uvarum. This was employed for the disruption of the HuATF1 gene, which encodes a putative alcohol acetyltransferase involved in acetate ester formation. We generated a synthetic marker gene consisting of the HuTEF1 promoter controlling a hygromycin resistance open reading frame (ORF). This new marker gene was used in disruption cassettes containing long-flanking (1000 bp) homology regions to the target locus. By increasing the antibiotic concentration, transformants were obtained in which both alleles of the putative HuATF1 gene were deleted in a diploid H. uvarum strain. Phenotypic characterisation including fermentation in Muller-Thurgau must showed that the null mutant produced significantly less acetate ester, particularly ethyl acetate. This study marks the first steps in the development of gene modification tools and paves the road for functional gene analyses of this yeast.
引用
收藏
页码:1 / 17
页数:16
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共 63 条
  • [1] Snails as Taxis for a Large Yeast Biodiversity
    Akan, Madina
    Michling, Florian
    Matti, Katrin
    Krause, Sinje
    Muno-Bender, Judith
    Wendland, Juergen
    [J]. FERMENTATION-BASEL, 2020, 6 (03):
  • [2] Yeast species associated with orange juice: Evaluation of different identification methods
    Arias, CR
    Burns, JK
    Friedrich, LM
    Goodrich, RM
    Parish, ME
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2002, 68 (04) : 1955 - 1961
  • [3] Growth promotion effect of Pichia guilliermondii in chilli and biocontrol potential of Hanseniaspora uvarum against Colletotrichum capsici causing fruit rot
    Basha, H.
    Ramanujam, B.
    [J]. BIOCONTROL SCIENCE AND TECHNOLOGY, 2015, 25 (02) : 185 - 206
  • [4] Dynamic behavior of Saccharomyces cerevisiae, Pichia kluyveri and Hanseniaspora uvarum during spontaneous and inoculated cocoa fermentations and their effect on sensory characteristics of chocolate
    Batista, Nadia Nara
    Ramos, Cintia Lacerda
    Ribeiro, Disney Dias
    Marques Pinheiro, Ana Carla
    Schwan, Rosane Freitas
    [J]. LWT-FOOD SCIENCE AND TECHNOLOGY, 2015, 63 (01) : 221 - 227
  • [5] Unraveling the Enzymatic Basis of Wine "Flavorome": A Phylo-Functional Study of Wine Related Yeast Species
    Belda, Ignacio
    Ruiz, Javier
    Alastruey-Izquierdo, Ana
    Navascues, Eva
    Marquina, Domingo
    Santos, Antonio
    [J]. FRONTIERS IN MICROBIOLOGY, 2016, 7
  • [6] Overexpression of RAD51 Enables PCR-Based Gene Targeting in Lager Yeast
    Bernardi, Beatrice
    Kayacan, Yeseren
    Akan, Madina
    Wendland, Juergen
    [J]. MICROORGANISMS, 2019, 7 (07):
  • [7] Development of headspace solid-phase microextraction-gas chromatography-mass spectrometry methodology for analysis of terpenoids in Madeira wines
    Câmara, JS
    Alves, MA
    Marques, JC
    [J]. ANALYTICA CHIMICA ACTA, 2006, 555 (02) : 191 - 200
  • [8] New and Redesigned pRS Plasmid Shuttle Vectors for Genetic Manipulation of Saccharomyces cerevisiae
    Chee, Mark K.
    Haase, Steven B.
    [J]. G3-GENES GENOMES GENETICS, 2012, 2 (05): : 515 - 526
  • [9] Genome sequencing and analysis of Kloeckera apiculata strain 34-9, a biocontrol agent against postharvest pathogens in citrus
    Chen, Kai
    Yang, Xiaoping
    Zheng, Feng
    Long, Chao-an
    [J]. GENES & GENOMICS, 2017, 39 (01) : 87 - 99
  • [10] The Fungal Aroma Gene ATF1 Promotes Dispersal of Yeast Cells through Insect Vectors
    Christiaens, Joaquin F.
    Franco, Luis M.
    Cools, Tanne L.
    De Meester, Luc
    Michiels, Jan
    Wenseleers, Tom
    Hassan, Bassem A.
    Yaksi, Emre
    Verstrepen, Kevin J.
    [J]. CELL REPORTS, 2014, 9 (02): : 425 - 432