Observing Nutrient Gradients, Gene Expression and Growth Variation Using the "Yeast Machine" Microfluidic Device
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Author:
Date:
2020-07-05
[Abstract] The natural environment of microbial cells like bacteria and yeast is often a complex community in which growth and internal organization reflect morphogenetic processes and interactions that are dependent on spatial position and time. While most of research is performed in simple homogeneous environments (e.g., bulk liquid cultures), which cannot capture full spatiotemporal community dynamics, studying biofilms or colonies is complex and usually does not give access to the spatiotemporal dynamics at single cell level. Here, we detail a protocol for generation of a microfluidic device, the “yeast machine”, with arrays of long monolayers of yeast colonies to advance the global understanding of how intercellular metabolic interactions affect the internal structure of colonies ...
[摘要] [摘要 ] 微生物细胞(如细菌和酵母菌)的自然环境通常是一个复杂的社区,在该社区中,生长和内部组织反映了形态发生过程和相互作用,这些过程和相互作用取决于空间位置和时间。虽然大多数研究是在无法捕获完整时空群落动态的简单同质环境(例如,大量液体培养)中进行的,但研究生物膜或菌落却很复杂,通常无法在单个细胞水平上获得时空动态。在这里,我们详细介绍了一种用于生成微流控设备(“酵母机器”)的协议,该协议带有酵母菌落的长单层阵列,以推进对细胞间代谢相互作用如何影响已定义和可定制的空间尺寸内菌落内部结构的全球了解。以酿酒酵母作为模型酵母系统,我们使用“酵母机器”通过追踪荧光标记的己糖转运蛋白来证明葡萄糖梯度的出现。我们进一步量化了菌落内生长速率的表达空间模式和葡萄糖可利用性调控的其他基因的表达。除此之外,我们显示出氨基酸的梯度也在菌落内形成,潜在地打开了类似的方法来研究许多其他营养物和代谢废物的梯度的时空形成。该方法将来可用于在与生态学和进化有关的单细胞分辨率和时标下,破译其他相同物种或更复杂的多物种系统中的远程代谢相互作用,细胞发育和形态发生之间的相互作用。
[背景 ] ...
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Generation of Fusarium graminearum Knockout Mutants by the Split-marker Recombination Approach
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Author:
Date:
2018-08-20
[Abstract] Fusarium graminearum is a destructive phytopathogen and shows an impressive metabolic diversity. Gene deletion is an important and useful approach for gene function study. Here we present a protocol for generating gene deletion mutant by applying “split-marker” deletion strategy (Catlett et al., 2003) with PEG-mediated protoplast transformation (Yuan et al., 2008; Martín, 2015).
[摘要] 禾谷镰刀菌是一种破坏性的植物病原体,具有令人印象深刻的代谢多样性。 基因缺失是基因功能研究的重要且有用的方法。 在这里,我们提出了一个协议,通过应用“分裂标记”删除策略(Catlett et al。,2003)与PEG介导的原生质体转化(Yuan 等。,2008;Martín,2015)。
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Perithecium Formation and Ascospore Discharge in Fusarium graminearum
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Author:
Date:
2018-08-05
[Abstract] The filamentous ascomycete Fusarium graminearum (previously also known as Gibberella zeae) is a phytopathogen of grain cereals, reducing crop yield and grain quality. The abilities of sexual reproduction organ-perithecium formation, ascospore formation and discharge are all essential characteristics relevant to F. graminearum disease cycle. Here, we present the details of the protocol to study perithecium formation and ascospore discharge in F. graminearum.
[摘要] 丝状子囊菌 Fusarium graminearum (以前也称为 Gibberella zeae )是谷物谷物的植物病原体,降低了作物产量和谷物质量。 有性生殖器官 - 外膜形成,子囊孢子形成和排出的能力都是与 F相关的基本特征。 禾本科疾病周期。 在这里,我们提出了研究 F中的perithecium形成和子囊孢子放电的协议的细节。菌。
【背景】子囊菌真菌 Fusarium graminearum 是小麦 Fusarium 头枯病和玉米赤霉病茎腐病的主要致病因子。这种真菌可以在定植的寄主植物表面产生有性子实体 - perithecia,在农作物碎片上过冬,并在明年流行时排出子囊孢子(Goswami和Kistler,2004)。受潮湿和温暖条件的影响,子囊孢子被强制从子囊中排出,并在气流中作为主要接种物空气传播。
这种真菌是同种异体的;大多数菌株可以很容易地在体外上生产胡萝卜琼脂上的perithecia(Trail and Common,2000)。已经报道了在 F的性发育期间的时间转录组学分析的微观研究和对perithecia发展的全面描述。 graminearum (Trail and Common,2000; Hallen et ...
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