The Na+/H+ antiporter Nhx1 controls vacuolar fusion indispensible for the life cycle in vitro and in vivo of a fungal insect pathogen
The Na+/H+ antiporter Nhx1 controls vacuolar fusion indispensible for the life cycle in vitro and in vivo of a fungal insect pathogen
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Na /H 逆向转运蛋白 Nhx1 控制液泡融合，这对于真菌昆虫病原体的体外和体内生命周期至关重要
DOI：
10.1111/1462-2920.13359
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发表时间：
2016
影响因子：
5.1
通讯作者：
Feng Ming-Guang
中科院分区：
文献类型：
--
作者：
Zhu Jin;Ying Sheng-Hua;Feng Ming-Guang
唯一的Na+/H+反向转运蛋白Nhx 1在丝状真菌中通常未被探索。我们对昆虫病原真菌球孢白僵菌（Beauveria bassiana）中的Nhx 1进行了鉴定。eGFP标记的Nhx 1融合体在小的点状结构中积累，推测为内体和跨高尔基体网络区室，在每个野生型细胞的间隔和管状空泡之间，用空泡特异性染料染色。nhx 1的缺失导致液泡中显著的酸化和严重的融合缺陷，这些液泡是碎片化的，与大的或管状的野生型液泡不同。该缺失还导致了气生分生孢子或淹没芽生孢子生产的急剧减少，以及营养生长比分生孢子萌发更严重的缺陷。Δ nhx 1突变体在生长过程中对高渗透压、热激和几种金属离子更加敏感，但其分生孢子表现出更强的UV-B耐受性。有趣的是，Δ nhx 1无法通过表皮穿透或血腔内注射感染模型昆虫，因为它在最小肉汤中产生的生物量和表皮降解酶要少得多，并且无法在昆虫血淋巴中形成芽生孢子。所有的变化完全或大部分恢复targetednhx 1互补。我们的研究结果提供了一个新的见解Nhx 1的一个不可缺少的不仅液泡融合，而且在体外和在vivoinB的生命周期。巴西纳。
The sole Na+/H+antiporter Nhx1 has been generally unexplored in filamentous fungi. We characterized Nhx1 in the entomopathogenic fungusBeauveria bassiana. An eGFP‐tagged Nhx1 fusion accumulated in small punctuate structures, presumably endosomal and trans‐Golgi network compartments, between septum and tubular vacuole of each wild‐type cell stained with a vacuole‐specific dye. Deletion ofnhx1resulted in significant acidification and severe fusion defect in vacuoles, which were fragmented and distinct from large or tubular wild‐type vacuoles. The deletion also caused a drastic reduction in aerial conidiation or submerged blastospore production and more severe defect in vegetative growth than in conidial germination. The Δnhx1mutant became more sensitive to high osmolarity, heat shock and several metal ions during growth but its conidia showed increased UV‐B tolerance. Intriguingly, Δnhx1was unable to infect a model insect through cuticle penetration or intrahaemocoel injection because it produced much less biomass and cuticle‐degrading enzymes in a minimal broth and failed to form blastospores in the insect haemolymph. All changes were completely or largely restored by targetednhx1complementation. Our results provide novel insight into an indispensability of Nhx1 for not only vacuolar fusion but also life cyclesin vitroandin vivoinB. bassiana.