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外被体调节香蕉枯萎病菌丝尖端和隔膜处水解酶分泌。

The Exocyst Regulates Hydrolytic Enzyme Secretion at Hyphal Tips and Septa in the Banana Fusarium Wilt Fungus Fusarium odoratissimum.

机构信息

State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, College of Plant Protection, Fujian Agriculture and Forestry Universitygrid.256111.0, Fuzhou, China.

College of Life Science, Fujian Agriculture and Forestry Universitygrid.256111.0, Fuzhou, China.

出版信息

Appl Environ Microbiol. 2021 Aug 11;87(17):e0308820. doi: 10.1128/AEM.03088-20.

Abstract

Hyphal polarized growth in filamentous fungi requires tip-directed secretion, while additional evidence suggests that fungal exocytosis for the hydrolytic enzyme secretion can occur at other sites in hyphae, including the septum. In this study, we analyzed the role of the exocyst complex involved in the secretion in the banana wilt fungal pathogen Fusarium odoratissimum. All eight exocyst components in not only localized to the tips ahead of the Spitzenkörper in growing hyphae but also localized to the outer edges of septa in mature hyphae. To further analyze the exocyst in , we attempted single gene deletion for all the genes encoding the eight exocyst components and only succeeded in constructing the gene deletion mutants for and ; we suspect that the other 6 exocyst components are encoded by essential genes. Deletion of or led to defects in vegetative growth, conidiation, and pathogenicity in . Notably, the deletion of resulted in decreased activities for endoglucosidase, filter paper enzymes, and amylase, while the loss of only led to a slight reduction in amylase activity. Septum-localized α-amylase (AmyB) was identified as the marker for septum-directed secretion, and we found that Exo70 is essential for the localization of AmyB to septa. Meanwhile the loss of Sec5 did not affect AmyB localization to septa but led to a higher accumulation of AmyB in cytoplasm. This suggested that while Exo70 and Sec5 both take part in the septum-directed secretion, the two play different roles in this process. The exocyst complex is a multisubunit tethering complex (MTC) for secretory vesicles at the plasma membrane and contains eight subunits, Sec3, Sec5, Sec6, Sec8, Sec10, Sec15, Exo70, and Exo84. While the exocyst complex is well defined in eukaryotes from yeast () to humans, the exocyst components in filamentous fungi show different localization patterns in the apical tips of hyphae, which suggests that filamentous fungi have evolved divergent strategies to regulate endomembrane trafficking. In this study, we demonstrated that the exocyst components in Fusarium odoratissimum are localized not only to the tips of growing hyphae but also to the outer edge of the septa in mature hyphae, suggesting that the exocyst complex plays a role in the regulation of septum-directed protein secretion in . We further found that Exo70 and Sec5 are required for the septum-directed secretion of α-amylase in but with different influences.

摘要

丝状真菌的菌丝极性生长需要指向顶端的分泌,而额外的证据表明,真菌胞吐作用可以在菌丝的其他部位发生,包括隔膜。在这项研究中,我们分析了参与分泌的外泌体复合物在香蕉枯萎病真菌病原体尖孢镰刀菌中的作用。不仅所有 8 种外泌体成分都定位于生长菌丝的 Spitzenkörper 前方的尖端,而且还定位于成熟菌丝的隔膜外边缘。为了进一步分析 中的外泌体,我们试图对编码 8 种外泌体成分的所有基因进行单基因缺失,但仅成功构建了 和 的基因缺失突变体;我们怀疑其他 6 种外泌体成分由必需基因编码。缺失 或 导致 在营养生长、产孢和致病性方面出现缺陷。值得注意的是,缺失 导致内切葡聚糖酶、滤纸酶和淀粉酶活性降低,而缺失 仅导致淀粉酶活性略有降低。定位于隔膜的 α-淀粉酶 (AmyB)被鉴定为隔膜定向分泌的标志物,我们发现 Exo70 是 AmyB 定位于隔膜所必需的。同时,Sec5 的缺失不影响 AmyB 定位于隔膜,但导致细胞质中 AmyB 的积累增加。这表明,虽然 Exo70 和 Sec5 都参与了隔膜定向分泌,但它们在这个过程中发挥着不同的作用。外泌体复合物是质膜上分泌囊泡的多亚基连接复合物 (MTC),包含 8 个亚基,即 Sec3、Sec5、Sec6、Sec8、Sec10、Sec15、Exo70 和 Exo84。虽然外泌体复合物在从酵母到人类的真核生物中已经得到很好的定义,但丝状真菌中的外泌体成分在菌丝顶端的定位模式不同,这表明丝状真菌已经进化出不同的策略来调节内膜运输。在这项研究中,我们证明了尖孢镰刀菌中的外泌体成分不仅定位于生长菌丝的尖端,而且还定位于成熟菌丝的隔膜外边缘,这表明外泌体复合物在调节 中隔膜定向蛋白分泌中发挥作用。我们进一步发现,Exo70 和 Sec5 是 中 α-淀粉酶隔膜定向分泌所必需的,但影响不同。

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