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Mixed yeast communities contribute to regionally distinct wine attributes. 混合酵母群落造就了葡萄酒与众不同的地方特色。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad005
Diana Lynne Hawkins, Jess Ryder, Soon A Lee, Katie Parish-Virtue, Bruno Fedrizzi, Matthew R Goddard, Sarah J Knight

There is evidence that vineyard yeast communities are regionally differentiated, but the extent to which this contributes to wine regional distinctiveness is not yet clear. This study represents the first experimental test of the hypothesis that mixed yeast communities-comprising multiple, region-specific, isolates, and species-contribute to regional wine attributes. Yeast isolates were sourced from uninoculated Pinot Noir fermentations from 17 vineyards across Martinborough, Marlborough, and Central Otago in New Zealand. New methodologies for preparing representative, mixed species inoculum from these significantly differentiated regional yeast communities in a controlled, replicable manner were developed and used to inoculate Pinot Noir ferments. A total of 28 yeast-derived aroma compounds were measured in the resulting wines via headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry. Yeast community region of origin had a significant impact on wine aroma, explaining ∼10% of the observed variation, which is in line with previous reports of the effects of region-specific Saccharomyces cerevisiae isolates on Sauvignon Blanc ferments. This study shows that regionally distinct, mixed yeast communities can modulate wine aroma compounds in a regionally distinct manner and are in line with the hypothesis that there is a microbial component to regional distinctiveness, or terroir, for New Zealand Pinot Noir.

有证据表明,葡萄园的酵母群落存在地区差异,但这种差异在多大程度上促进了葡萄酒的地区独特性尚不清楚。本研究首次对混合酵母群落--包括多种地区特异性分离物和物种--有助于葡萄酒地区属性的假设进行了实验测试。酵母分离物来自新西兰马丁堡、马尔堡和中奥塔哥地区 17 个葡萄园的未接种黑比诺发酵物。我们开发了新方法,以可控、可复制的方式从这些差异显著的地区酵母群落中制备具有代表性的混合物种接种物,并将其用于黑比诺发酵接种。通过顶空固相微萃取和气相色谱-质谱联用技术,在酿制的葡萄酒中测定了 28 种酵母衍生香气化合物。酵母群落的原产地对葡萄酒香气有显著影响,可解释观察到的变化的 10%,这与之前关于特定地区的酿酒酵母分离株对长相思发酵的影响的报道一致。这项研究表明,不同地区的混合酵母群落能以不同地区的方式调节葡萄酒的香气化合物,这与新西兰黑比诺的地区特色或风土中存在微生物成分的假设是一致的。
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引用次数: 0
Production of single cell oil by two novel nonconventional yeast strains of Curvibasidium sp. isolated from medicinal lichen. 两株从药用地衣中分离的曲子菌属新酵母株生产单细胞油的研究。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad026
Long Bai, Cheng Cheng, Meng-Lin Sun, Jun Li, Yue Zou, Quanyu Zhao, Xin-Qing Zhao

Oleaginous yeasts utilize renewable resources to produce lipids, which benefits sustainable development, and it is of great interest to screen robust lipid producers. Curvibasidium sp. belongs to nonconventional yeast that are very limitedly studied. Here, two cold-adaptive strains of Curvibasidium sp., namely, Y230 and Y231, isolated from the medicinal lichen Usnea diffracta were investigated for their potential in lipid production. Genome mining of Curvibasidium sp. Y231 was performed, and the special features related to fatty acid biosynthesis were revealed. Glucose, xylose, and glycerol were tested as sole carbon sources for yeast cell growth and lipid production. The total lipid contents of Curvibasidium sp. Y230 and Y231 range from 38.43% to 54.62% of the cell dry cell weight at 20°C, and glucose is the optimal carbon source. These results indicate that the Curvibasidium sp. strains are promising for sustainable lipid production. Our study provides basis for exploration of lichen-derived strains for biotechnological applications, and also benefits utilization of other nonconventional yeasts for sustainable production based on genome-based studies.

产油酵母利用可再生资源生产油脂,有利于油脂的可持续发展,因此筛选优质产脂菌具有重要意义。曲担子酵母属一种研究非常有限的非常规酵母。本研究从药用地衣Usnea diffracta中分离出两株冷适应性菌株Curvibasidium sp.,即Y230和Y231,研究其产脂能力。对曲子菌(Curvibasidium sp. Y231)进行了基因组挖掘,揭示了与脂肪酸生物合成相关的特殊特征。葡萄糖、木糖和甘油被测试为酵母细胞生长和脂质生产的唯一碳源。在20℃条件下,曲子菌Y230和Y231的总脂质含量为细胞干重的38.43% ~ 54.62%,葡萄糖为最佳碳源。这些结果表明,曲担子菌具有持续产脂的潜力。本研究为探索地衣衍生菌株的生物技术应用提供了基础,也有利于利用其他基于基因组研究的非常规酵母进行可持续生产。
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引用次数: 0
Polymorphisms of rDNA genes in Cyberlindnera yeast suggest birth-and-death evolution events. Cyberlindnera 酵母菌中 rDNA 基因的多态性提示了生死进化事件。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad032
Rodolfo Bizarria, Tatiane de Castro Pietrobon, Henrique Ferreira, Andre Rodrigues

In eukaryotes, the ribosome machinery is encoded by repeats of the ribosomal RNA genes: 26/28S, 18S, 5.8S, and 5S, structured in tandem arrays and frequently homogenized within a genome. This homogenization is thought to be driven by concerted evolution, evolving as a unit, which contributes to its target as the species barcode in modern taxonomy. However, high heterogeneity of rDNA genes has been reported, including in Saccharomycotina yeasts. Here, we describe the polymorphisms and heterogeneity of D1/D2 domains (26S rRNA) and the intergenic transcribed spacer of a new yeast species with affinities to the genus Cyberlindnera and their evolution. Both regions are not homogenized, failing the prediction of concerted evolution. Phylogenetic network analysis of cloned sequences revealed that Cyberlindnera sp. rDNAs are diverse and evolved by reticulation rather than by bifurcating tree evolution model. Predicted rRNA secondary structures also confirmed structural differences, except for some conserved hairpin loops. We hypothesize that some rDNA is inactive within this species and evolves by birth-and-death rather than concerted evolution. Our findings propel further investigation into the evolution of rDNA genes in yeasts.

在真核生物中,核糖体机器是由核糖体 RNA 基因的重复序列编码的:26/28S、18S、5.8S 和 5S,呈串联阵列结构,在基因组内经常同质化。这种同质化被认为是由协同进化驱动的,是作为一个单元进化的,这有助于在现代分类学中将其作为物种条形码的目标。然而,rDNA 基因的高度异质性已被报道,包括在酵母菌中。在这里,我们描述了一个与 Cyberlindnera 属有亲缘关系的酵母新种的 D1/D2 域(26S rRNA)和基因间转录间隔的多态性和异质性及其进化。这两个区域没有同质化,无法预测协同进化。对克隆序列的系统进化网络分析显示,Cyberlindnera sp.的rDNA具有多样性,是通过网状进化而非分叉树进化模式进化而来的。预测的 rRNA 二级结构也证实了结构上的差异,除了一些保守的发夹环之外。我们推测,一些 rDNA 在该物种内不活跃,是通过生死进化而非协同进化的。我们的发现推动了对酵母中 rDNA 基因进化的进一步研究。
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引用次数: 0
Elevated energy costs of biomass production in mitochondrial respiration-deficient Saccharomyces cerevisia. 线粒体呼吸缺陷酿酒酵母生物质生产的能源成本升高。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad008
Pranas Grigaitis, Samira L van den Bogaard, Bas Teusink

Microbial growth requires energy for maintaining the existing cells and producing components for the new ones. Microbes therefore invest a considerable amount of their resources into proteins needed for energy harvesting. Growth in different environments is associated with different energy demands for growth of yeast Saccharomyces cerevisiae, although the cross-condition differences remain poorly characterized. Furthermore, a direct comparison of the energy costs for the biosynthesis of the new biomass across conditions is not feasible experimentally; computational models, on the contrary, allow comparing the optimal metabolic strategies and quantify the respective costs of energy and nutrients. Thus in this study, we used a resource allocation model of S. cerevisiae to compare the optimal metabolic strategies between different conditions. We found that S. cerevisiae with respiratory-impaired mitochondria required additional energetic investments for growth, while growth on amino acid-rich media was not affected. Amino acid supplementation in anaerobic conditions also was predicted to rescue the growth reduction in mitochondrial respiratory shuttle-deficient mutants of S. cerevisiae. Collectively, these results point to elevated costs of resolving the redox imbalance caused by de novo biosynthesis of amino acids in mitochondria. To sum up, our study provides an example of how resource allocation modeling can be used to address and suggest explanations to open questions in microbial physiology.

微生物的生长需要能量来维持现有的细胞,并为新的细胞产生成分。因此,微生物将大量资源投入到能量收集所需的蛋白质中。不同环境下的生长与酵母生长所需的能量不同有关,尽管交叉条件的差异仍然不清楚。此外,直接比较不同条件下新生物质生物合成的能源成本在实验上是不可实现的;相反,计算模型可以比较最佳代谢策略,并量化各自的能量和营养成本。因此,在本研究中,我们使用酿酒酵母资源分配模型来比较不同条件下的最佳代谢策略。我们发现线粒体呼吸受损的酿酒酵母需要额外的能量投入来生长,而在富含氨基酸的培养基上的生长不受影响。在厌氧条件下补充氨基酸也被预测可以挽救酿酒酵母线粒体呼吸梭缺陷突变体的生长减少。总的来说,这些结果表明解决线粒体中氨基酸从头生物合成引起的氧化还原不平衡的成本增加。总之,我们的研究提供了一个例子,说明如何使用资源分配模型来解决和解释微生物生理学中的开放性问题。
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引用次数: 0
Zuo1, a ribosome-associated J protein, is involved in glucose repression in Saccharomyces cerevisiae. Zuo1是一种核糖体相关的J蛋白,参与酿酒酵母的葡萄糖抑制。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad038
Yoichi Yamada, Atsuki Shiroma, Suguru Hirai, Jun Iwasaki

In Saccharomyces cerevisiae, the J-protein Zuo1 and the nonconventional Hsp70 homologue Ssz1 stimulate the ATPase activity of the chaperone proteins Ssb1 and Ssb2 (Ssb1/2), which are associated with the ribosomes. The dephosphorylation of sucrose nonfermenting 1 (Snf1) on Thr210 is required for glucose repression. The Ssb1/2 and 14-3-3 proteins Bmh1 and Bmh2 appear to be responsible for the dephosphorylation of Snf1 on Thr210 and glucose repression. Here, we investigated the role of Zuo1 in glucose repression. The zuo1∆ strain as well as the ssb1∆ssb2∆ strain exhibited a glucose-specific growth defect during logarithmic growth on glucose. Many of the respiratory chain genes examined were statistically significantly upregulated, but less than 2-fold, in the zuo1∆ strain as well as in the ssb1∆ssb2∆ strain on glucose. In addition, excessive phosphorylation of Snf1 on Thr210 was observed in the zuo1∆ strain as well as in the ssb1∆ssb2∆ strain in the presence of glucose. The mRNA levels of SSB1/2 and BMH1 were statistically significantly reduced by approximately 0.5- to 0.8-fold relative to the wild-type level in the zuo1∆ strain on glucose. These results suggest that Zuo1 is responsible for glucose repression, possibly by increasing the mRNA levels of SSB1/2 and BMH1 during growth on glucose.

在酿酒酵母中,j蛋白Zuo1和非常规的Hsp70同源物Ssz1刺激与核糖体相关的伴侣蛋白Ssb1和Ssb2 (Ssb1/2)的atp酶活性。蔗糖非发酵1 (Snf1)在Thr210上的去磷酸化是葡萄糖抑制所必需的。Ssb1/2和14-3-3蛋白Bmh1和Bmh2似乎负责Snf1对Thr210的去磷酸化和葡萄糖抑制。在这里,我们研究了Zuo1在葡萄糖抑制中的作用。在葡萄糖的对数生长过程中,zuo1∆菌株和ssb1∆ssb2∆菌株表现出葡萄糖特异性生长缺陷。在葡萄糖作用下的zuo1∆菌株和ssb1∆ssb2菌株中,许多被检测的呼吸链基因在统计学上显著上调,但低于2倍。此外,在葡萄糖存在的情况下,在zuo1∆菌株和ssb1∆ssb2∆菌株中观察到Thr210上Snf1的过度磷酸化。葡萄糖zuo1∆菌株的SSB1/2和BMH1 mRNA水平较野生型降低约0.5 ~ 0.8倍,差异有统计学意义。这些结果表明,Zuo1可能通过增加葡萄糖生长过程中SSB1/2和BMH1的mRNA水平来抑制葡萄糖。
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引用次数: 0
Non-saccharomyces yeast probiotics: revealing relevance and potential. 非酿酒酵母益生菌:揭示相关性和潜力。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad041
Silvia Cristina Vergara, María José Leiva, María Victoria Mestre, Fabio Vazquez, María Cristina Nally, Yolanda Paola Maturano

Non-Saccharomyces yeasts are unicellular eukaryotes that play important roles in diverse ecological niches. In recent decades, their physiological and morphological properties have been reevaluated and reassessed, demonstrating the enormous potential they possess in various fields of application. Non-Saccharomyces yeasts have gained relevance as probiotics, and in vitro and in vivo assays are very promising and offer a research niche with novel applications within the functional food and nutraceutical industry. Several beneficial effects have been described, such as antimicrobial and antioxidant activities and gastrointestinal modulation and regulation functions. In addition, several positive effects of bioactive compounds or production of specific enzymes have been reported on physical, mental and neurodegenerative diseases as well as on the organoleptic properties of the final product. Other points to highlight are the multiomics as a tool to enhance characteristics of interest within the industry; as well as microencapsulation offer a wide field of study that opens the niche of food matrices as carriers of probiotics; in turn, non-Saccharomyces yeasts offer an interesting alternative as microencapsulating cells of various compounds of interest.

非酿酒酵母是单细胞真核生物,在不同的生态位中发挥着重要作用。近几十年来,人们对它们的生理和形态特性进行了重新评估,证明了它们在各个应用领域中具有巨大的潜力。非酿酒酵母作为益生菌已经获得了相关性,体外和体内检测非常有前景,并在功能性食品和营养品行业提供了新的应用研究领域。已经描述了几种有益的效果,如抗菌和抗氧化活性以及胃肠道调节和调节功能。此外,据报道,生物活性化合物或特定酶的产生对身体、精神和神经退行性疾病以及最终产品的感官特性有一些积极影响。其他需要强调的是,多元组学是增强行业内兴趣特征的工具;以及微胶囊化提供了广泛的研究领域,打开了食品基质作为益生菌载体的利基市场;反过来,非酿酒酵母作为各种感兴趣化合物的微胶囊化细胞提供了一种有趣的替代品。
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引用次数: 0
Yca1 metacaspase: diverse functions determine how yeast live and let die. Yca1元天冬酶:多种功能决定酵母如何生存和死亡。
IF 2.4 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad022
Darren K Lam, Gavin Sherlock

The Yca1 metacaspase was discovered due to its role in the regulation of apoptosis in Saccharomyces cerevisiae. However, the mechanisms that drive apoptosis in yeast remain poorly understood. Additionally, Yca1 and other metacaspase proteins have recently been recognized for their involvement in other cellular processes, including cellular proteostasis and cell cycle regulation. In this minireview, we outline recent findings on Yca1 that will enable the further study of metacaspase multifunctionality and novel apoptosis pathways in yeast and other nonmetazoans. In addition, we discuss advancements in high-throughput screening technologies that can be applied to answer complex questions surrounding the apoptotic and nonapoptotic functions of metacaspase proteins across a diverse range of species.

Yca1 元天冬酶之所以被发现,是因为它在调控酿酒酵母的细胞凋亡中发挥了作用。然而,人们对酵母细胞凋亡的驱动机制仍然知之甚少。此外,Yca1 和其他元aspase 蛋白最近也被认为参与了其他细胞过程,包括细胞蛋白稳态和细胞周期调控。在本小视图中,我们概述了有关 Yca1 的最新发现,这些发现将有助于进一步研究元aspase 的多功能性以及酵母和其他非甲动物的新型凋亡途径。此外,我们还讨论了高通量筛选技术的进展,这些技术可用于回答围绕元aspase 蛋白在不同物种中的凋亡和非凋亡功能的复杂问题。
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引用次数: 0
Genomic and functional features of yeast species in Korean traditional fermented alcoholic beverage and soybean products. 韩国传统发酵酒精饮料和豆制品中酵母菌种的基因组和功能特征。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foac066
Da Min Jeong, Hyeon Jin Kim, Min-Seung Jeon, Su Jin Yoo, Hye Yun Moon, Eun-Joo Jeon, Che Ok Jeon, Seong-Il Eyun, Hyun Ah Kang

In this review, we describe the genomic and physiological features of the yeast species predominantly isolated from Nuruk, a starter for traditional Korean rice wines, and Jang, a traditional Korean fermented soy product. Nuruk and Jang have several prevalent yeast species, including Saccharomycopsis fibuligera, Hyphopichia burtonii, and Debaryomyces hansenii complex, which belong to the CUG clade showing high osmotic tolerance. Comparative genomics revealed that the interspecies hybridization within yeast species for generating heterozygous diploid genomes occurs frequently as an evolutional strategy in the fermentation environment of Nuruk and Jang. Through gene inventory analysis based on the high-quality reference genome of S. fibuligera, new genes involved in cellulose degradation and volatile aroma biosynthesis and applicable to the production of novel valuable enzymes and chemicals can be discovered. The integrated genomic and transcriptomic analysis of Hyphopichia yeasts, which exhibit strong halotolerance, provides insights into the novel mechanisms of salt and osmo-stress tolerance for survival in fermentation environments with a low-water activity and high-concentration salts. In addition, Jang yeast isolates, such as D. hansenii, show probiotic potential for the industrial application of yeast species beyond fermentation starters to diverse human health sectors.

在这篇综述中,我们描述了主要从韩国传统米酒的发酵剂 Nuruk 和韩国传统发酵豆制品 Jang 中分离出来的酵母菌种的基因组和生理特征。Nuruk 和 Jang 有几种常见的酵母菌,包括 Saccharomycopsis fibuligera、Hyphopichia burtonii 和 Debaryomyces hansenii complex,它们属于 CUG 支系,具有高渗透耐受性。比较基因组学发现,在努鲁克和张氏发酵环境中,作为一种进化策略,酵母菌种间杂交以产生杂合二倍体基因组的现象经常发生。通过基于 S. fibuligera 高质量参考基因组的基因清单分析,可以发现参与纤维素降解和挥发性香气生物合成的新基因,这些基因可用于生产新型有价值的酶和化学品。通过对耐盐性强的 Hyphopichia 酵母菌的基因组和转录组的综合分析,可以深入了解其在低水活性和高浓度盐的发酵环境中生存的耐盐和耐渗透压的新机制。此外,张氏酵母分离物(如 D. hansenii)显示出益生菌的潜力,可将酵母物种的工业应用从发酵起动器扩展到各种人类健康领域。
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引用次数: 1
Influence of different Lachancea thermotolerans strains in the wine profile in the era of climate challenge. 气候变化下不同耐温葡萄品种对葡萄酒品质的影响。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foac062
Javier Vicente, Niina Kelanne, Lydia Rodrigo-Burgos, Eva Navascués, Fernando Calderón, Antonio Santos, Domingo Marquina, Baoru Yang, Santiago Benito

The study performed sequential fermentations of red grape juice using several strains of Lachancea thermotolerans and one strain of Saccharomyces cerevisiae. Due to the new conditions imposed by climate change, wine acidity must be affected as well as the volatile profile. Non-Saccharomyces yeasts such as L. thermotolerans are real alternatives to soften the impact of climate change in winemaking. The L. thermotolerans strains included three commercially available strains and two wine-related natural isolates. L. thermotolerans showed significant statistical differences in basic chemical parameters such as lactic acid, malic acid, or ethanol concentrations as well as in the volatile profile. S. cerevisiae clearly produced some volatile compounds in higher amounts than the studied L. thermotolerans strains while others showed the opposite effect. Sequential fermentations involving any of the studied strains of L. thermotolerans with S. cerevisiae showed an increased volatile profile compared to the S. ceresisiae single fermentation, highlighting the synergic effect between the studied species.

本研究利用几株耐高温葡萄球菌和一株酿酒酵母菌对红葡萄汁进行了连续发酵。由于气候变化带来的新条件,葡萄酒的酸度必然受到影响,其挥发性也会受到影响。非酵母菌酵母菌,如耐高温酵母菌,是缓解气候变化对酿酒影响的真正替代品。耐温L.菌株包括3个市售菌株和2个与葡萄酒相关的天然菌株。耐温菌在乳酸、苹果酸或乙醇浓度等基本化学参数以及挥发性谱上表现出显著的统计学差异。酿酒葡萄球菌明显比所研究的耐高温葡萄球菌产生更多的挥发性化合物,而其他菌株则表现出相反的效果。与酿酒酵母单次发酵相比,任何一种耐热L.菌株与酿酒酵母的连续发酵都显示出更高的挥发性,这突出了所研究物种之间的协同效应。
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引用次数: 1
CRISPR-Cas9 engineering in the hybrid yeast Zygosaccharomyces parabailii can lead to loss of heterozygosity in target chromosomes. 在杂交酵母Zygosaccharomyces parabailii中进行CRISPR-Cas9工程可导致靶染色体的杂合性丧失。
IF 3.2 4区 生物学 Q2 Immunology and Microbiology Pub Date : 2023-01-04 DOI: 10.1093/femsyr/foad036
Pooja Jayaprakash, Liliane Barroso, Matteo Vajente, Letizia Maestroni, Edward J Louis, John P Morrissey, Paola Branduardi

The hybrid yeast Zygosaccharomyces parabailii holds potential as a cell factory mainly because of its robustness in withstanding stressors that often characterize bio-based processes. However, a complex genome and a lack of gene editing tools hinder the capacity to engineer this yeast. In this work, we developed a CRISPR-Cas9 gene editing system for Z. parabailii that allows simultaneous disruption or deletion of both alleles of a gene. We evaluated four different gRNA expression systems consisting of combinations of tRNAs, tRNA and ribozyme or ribozymes as self-cleaving flanking elements and established that the most efficient systems used an RNA Pol II promoter followed by a 5'tRNA flanking the gRNA. This gRNA system was then used to construct a strain of Z. parabailii in which both alleles of DNL4 were inactivated and so relied on homologous recombination to repair double-stranded breaks. Our system can be used for gene inactivation in a wild-type strain and precise deletion with marker insertion in a dnl4 mutant. In some cases, we observed inter-chromosomal recombination around the site of the DSB that could cause loss of heterozygosity through gene conversion or deletion. Although an additional aspect that needs to be monitored during strain engineering, this phenomenon also offers opportunities to explore genome plasticity in hybrid yeasts.

杂交酵母Zygosaccharomyces parabailii具有作为细胞工厂的潜力,主要是因为它在承受通常表征生物基过程的应激源方面具有稳健性。然而,复杂的基因组和缺乏基因编辑工具阻碍了改造这种酵母的能力。在这项工作中,我们为Z. parabailii开发了一种CRISPR-Cas9基因编辑系统,该系统允许同时破坏或删除基因的两个等位基因。我们评估了四种不同的gRNA表达系统,包括tRNA、tRNA和核酶或核酶作为自切割侧翼元件的组合,并确定了最有效的系统使用RNA Pol II启动子,然后是gRNA侧翼的5'tRNA。然后利用该gRNA系统构建了一株双链断裂的Z. parabailii菌株,其中DNL4的两个等位基因都失活,因此依靠同源重组来修复双链断裂。我们的系统可以用于野生型菌株的基因失活和dn14突变体的精确缺失与标记插入。在某些情况下,我们观察到DSB位点周围的染色体间重组可能通过基因转换或缺失导致杂合性丧失。虽然这是菌株工程中需要监测的另一个方面,但这种现象也为探索杂交酵母的基因组可塑性提供了机会。
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引用次数: 0
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