Abstract：
Nosema bombycis, an obligate intracellular parasite, is a single-celled eukaryote known to infect various tissues of silkworms, leading to the manifestation of pebrine. Trehalase, a glycosidase responsible for catalyzing the hydrolysis of trehalose into two glucose molecules, assumes a crucial role in thermal stress tolerance, dehydration, desiccation stress, and asexual development. Despite its recognized importance in these processes, the specific role of trehalase in N. bombycis remains uncertain. This investigation focused on exploring the functions of trehalase 3 in N. bombycis (NbTre3). Immunofluorescence analysis of mature (dormant) spores indicated that NbTre3 primarily localizes to the spore membrane or spore wall, suggesting a potential involvement in spore germination. Reverse transcription-quantitative polymerase chain reaction results indicated that the transcriptional level of NbTre3 peaked at 6 h post N. bombycis infection, potentially contributing to energy storage for proliferation. Throughout the life cycle of N. bombycis within the host cell, NbTre3 was detected in sporoplasm during the proliferative stage rather than the sporulation stage. RNA interference experiments revealed a substantial decrease in the relative transcriptional level of NbTre3, accompanied by a certain reduction in the relative transcriptional level of Nb16S rRNA. These outcomes suggest that NbTre3 may play a role in the proliferation of N. bombycis. The application of the His pull-down technique identified 28 proteins interacting with NbTre3, predominantly originating from the host silkworm. This finding implies that NbTre3 may participate in the metabolism of the host cell, potentially utilizing the host cell\'s energy resources.
摘要：
Nosemabombycis,一种专性细胞内寄生虫，是一种单细胞真核生物，可以感染蚕的各种组织，导致peparin的表现。海藻糖酶,一种糖苷酶，负责催化海藻糖水解成两个葡萄糖分子，在热应力耐受性中起着至关重要的作用，脱水,干燥应力，和无性发育。尽管它在这些进程中具有公认的重要性，海藻糖酶在烟草中的具体作用仍不确定。这项研究的重点是探索海藻糖酶3在家蚕原虫（NbTre3）中的功能。对成熟（休眠）孢子的免疫荧光分析表明，NbTre3主要位于孢子膜或孢子壁，提示可能参与孢子萌发。逆转录定量聚合酶链反应结果表明，NbTre3的转录水平在N.bombycis感染后6h达到峰值，可能有助于为扩散储存能量。在宿主细胞内的整个生命周期中，在增殖阶段而不是孢子形成阶段的孢子浆中检测到NbTre3。RNA干扰实验表明，NbTre3的相对转录水平显着降低，同时Nb16SrRNA的相对转录水平也有一定降低。这些结果表明，NbTre3可能在家蚕梭菌的增殖中起作用。His下拉技术的应用鉴定了与NbTre3相互作用的28种蛋白质，主要来自宿主家蚕。这一发现暗示NbTre3可能参与宿主细胞的代谢，潜在地利用宿主细胞的能量资源。
