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Functional Analysis of the α-1,3-Glucan Synthase Genes agsA and agsB in Aspergillus nidulans: AgsB Is the Major α-1,3-Glucan Synthase in This Fungus

机译:nidulans中α-1,3-葡聚糖合酶基因agsA和agsB的功能分析:AgsB是该真菌中主要的α-1,3-葡聚糖合酶

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摘要

Although α-1,3-glucan is one of the major cell wall polysaccharides in filamentous fungi, the physiological roles of α-1,3-glucan remain unclear. The model fungus Aspergillus nidulans possesses two α-1,3-glucan synthase (AGS) genes, agsA and agsB. For functional analysis of these genes, we constructed several mutant strains in A. nidulans: agsA disruption, agsB disruption, and double-disruption strains. We also constructed several CagsB strains in which agsB expression was controlled by the inducible alcA promoter, with or without the agsA-disrupting mutation. The agsA disruption strains did not show markedly different phenotypes from those of the wild-type strain. The agsB disruption strains formed dispersed hyphal cells under liquid culture conditions, regardless of the agsA genetic background. Dispersed hyphal cells were also observed in liquid culture of the CagsB strains when agsB expression was repressed, whereas these strains grew normally in plate culture even under the agsB-repressed conditions. Fractionation of the cell wall based on the alkali solubility of its components, quantification of sugars, and 13C-NMR spectroscopic analysis revealed that α-1,3-glucan was the main component of the alkali-soluble fraction in the wild-type and agsA disruption strains, but almost no α-1,3-glucan was found in the alkali-soluble fraction derived from either the agsB disruption strain or the CagsB strain under the agsB-repressed conditions, regardless of the agsA genetic background. Taken together, our data demonstrate that the two AGS genes are dispensable in A. nidulans, but that AgsB is required for normal growth characteristics under liquid culture conditions and is the major AGS in this species.
机译:尽管α-1,3-葡聚糖是丝状真菌中主要的细胞壁多糖之一,但是α-1,3-葡聚糖的生理作用仍然不清楚。模型真菌构巢曲霉(Aspergillus nidulans)具有两个α-1,3-葡聚糖合酶(AGS)基因agsA和agsB。为了对这些基因进行功能分析,我们在构巢曲霉中构建了几种突变菌株:agsA破坏,agsB破坏和双破坏菌株。我们还构建了几种CagsB菌株,其中agsB的表达受可诱导的alcA启动子控制,带有或不带有破坏agsA的突变。 agsA破坏菌株没有显示出与野生型菌株明显不同的表型。无论agsA的遗传背景如何,agsB破坏菌株在液体培养条件下均会形成分散的菌丝细胞。当抑制agsB表达时,在CagsB菌株的液体培养物中也观察到分散的菌丝细胞,而这些菌株即使在受agsB抑制的条件下,在平板培养中也能正常生长。根据其成分的碱溶解度,糖的定量和 13 C-NMR光谱分析对细胞壁进行分级分离,结果表明,α-1,3-葡聚糖是该碱溶性物质的主要成分野生型和agsA破坏菌株中的部分,但在 agsB 下从agsB破坏菌株或CagsB菌株衍生的碱溶部分中几乎没有发现α-1,3-葡聚糖。抑制条件,无论 agsA 遗传背景如何。两者合计,我们的数据表明这两个AGS基因在 A中是不可缺少的。 nidulans ,但AgsB是液体培养条件下正常生长特性所必需的,并且是该物种中的主要AGS。

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