Flb 蛋白家族在黑曲霉生命周期中的作用

Xiaoyi Chen, Juan P. Moran Torres, Han A. B. Wösten
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摘要

基因 flbA-E 参与黑曲霉的孢子形成和无性生殖。这些基因中的任何一个失活都会导致孢子形成延迟甚至消失的蓬松表型。此前,通过使黑曲霉中的 flbA 失活,获得了一种无孢子表型,这种表型伴随着裂解、细胞壁变薄和分泌组复杂性增加。在此,我们进一步研究了黑曲霉的 flb 基因的作用。菌株 ΔflbA、ΔflbB 和 ΔflbE显示生物量形成增加,而flbA-D的失活则减少甚至取消了分生孢子的形成。菌株 ΔflbA 对 H2O2、DTT 以及细胞壁完整性应激化合物 SDS 和刚果红(CR)更敏感。此外,ΔflbC 对 SDS 更为敏感,而 ΔflbB、ΔflbD 和 ΔflbE则对 CR 更为敏感。另一方面,flbE 失活会增加对 H2O2 的抵抗力。当Δflb菌株在木糖上生长时,酶的分泌受到影响。ΔflbE 菌株的木聚糖酶、纤维素酶和淀粉酶分泌减少。另一方面,ΔflbA菌落外围的淀粉酶分泌减少,但其中心却没有减少,而ΔflbB菌落中心的淀粉酶分泌增加,但其外围却没有增加。flbC和flbD的失活也会影响带状纤维素酶和淀粉酶的活性。黑僵菌的 Flb 蛋白家族在生物量形成、孢子形成、应激反应和蛋白质分泌方面发挥着重要作用。
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The role of the Flb protein family in the life cycle of Aspergillus niger

Genes flbA-E are involved in sporulation and vegetative growth in Aspergillus nidulans. Inactivation of either of these genes results in a fluffy phenotype with delayed or even abolished sporulation. Previously, a non-sporulating phenotype was obtained by inactivating flbA in Aspergillus niger, which was accompanied by lysis, thinner cell walls, and an increased secretome complexity. Here, we further studied the role of the flb genes of A. niger. Strains ΔflbA, ΔflbB and ΔflbE showed increased biomass formation, while inactivation of flbA-D reduced, or even abolished, formation of conidia. Strain ΔflbA was more sensitive to H2O2, DTT, and the cell wall integrity stress compounds SDS and Congo Red (CR). Also, ΔflbC was more sensitive to SDS, while ΔflbB, ΔflbD, and ΔflbE were more sensitive to CR. On the other hand, inactivation of flbE increased resistance to H2O2. Enzyme secretion was impacted when the Δflb strains were grown on xylose. Strain ΔflbE showed reduced xylanase, cellulase and amylase secretion. On the other hand, amylase secretion at the periphery of the ΔflbA colony was reduced but not in its center, while secretion of this enzyme was increased in the center of the ΔflbB colony but not at its periphery. Inactivation of flbC and flbD also impacted zonal cellulase and amylase activity. Together, the Flb protein family of A. niger function in biomass formation, sporulation, stress response, and protein secretion.

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