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Bioactive Excreted/Secreted Products of Entomopathogenic Nematode Heterorhabditis bacteriophora Inhibit the Phenoloxidase Activity during the Infection.
Insects ( IF 3 ) Pub Date : 2020-06-05 , DOI: 10.3390/insects11060353
Sara Eliáš 1 , Jana Hurychová 1 , Duarte Toubarro 2 , Jorge Frias 2 , Martin Kunc 1 , Pavel Dobeš 1 , Nelson Simões 2 , Pavel Hyršl 1
Affiliation  

Entomopathogenic nematodes (EPNs) are efficient insect parasites, that are known for their mutualistic relationship with entomopathogenic bacteria and their use in biocontrol. EPNs produce bioactive molecules referred to as excreted/secreted products (ESPs), which have come to the forefront in recent years because of their role in the process of host invasion and the modulation of its immune response. In the present study, we confirmed the production of ESPs in the EPN Heterorhabditis bacteriophora, and investigated their role in the modulation of the phenoloxidase cascade, one of the key components of the insect immune system. ESPs were isolated from 14- and 21-day-old infective juveniles of H. bacteriophora, which were found to be more virulent than newly emerged nematodes, as was confirmed by mortality assays using Galleria mellonella larvae. The isolated ESPs were further purified and screened for the phenoloxidase-inhibiting activity. In these products, a 38 kDa fraction of peptides was identified as the main candidate source of phenoloxidase-inhibiting compounds. This fraction was further analyzed by mass spectrometry and the de novo sequencing approach. Six peptide sequences were identified in this active ESP fraction, including proteins involved in ubiquitination and the regulation of a Toll pathway, for which a role in the regulation of insect immune response has been proposed in previous studies.

中文翻译:

昆虫病原线虫细菌性异型杆菌的生物活性分泌/分泌产物在感染过程中抑制酚氧化酶活性。

昆虫病原线虫(EPN)是有效的昆虫寄生虫,以其与昆虫病原菌的相互关系及其在生物防治中的应用而闻名。EPNs产生称为排泄/分泌产物(ESP)的生物活性分子,由于它们在宿主入侵过程中的作用及其免疫应答的调节作用,近年来已成为最前沿。在本研究中,我们确认了EPNs在EPN细菌性杂种优势细菌中的产生,并研究了它们在调节昆虫免疫系统关键成分之一的酚氧化酶级联反应中的作用。从14和21天大的H. bacteriophora感染少年中分离出ESP蠕虫被发现比新出现的线虫更具杀伤力,这已通过使用圆顶虫幼虫进行的死亡率分析得到了证实。分离的ESP被进一步纯化,并筛选其抑制酚氧化酶的活性。在这些产品中,鉴定出38 kDa的肽段是酚氧化酶抑制化合物的主要候选来源。通过质谱法和从头测序法进一步分析该级分。在该活性ESP馏分中鉴定出六个肽序列,包括涉及泛素化和Toll途径调控的蛋白质,在先前的研究中已提出了在昆虫免疫应答调控中的作用。
更新日期:2020-06-05
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