β‐1,3‐glucan recognition proteins (βGRPs) function as pattern recognition receptors in the innate immune response against invading pathogens. In the present study, we obtain full‐length cDNA clones for two novel putative βGRPs: TpβGRPc and TpβGRPd from the ghost moth Thitarodes pui (Lepidoptera: Hepialidae). Phylogenetic analysis shows a small distinct lineage, βGRP clade 4, consisting of T. pui βGRPs including TpβGRPa and TpβGRPb that have been identified previously. TpβGRPc and TpβGRPd, comprising 488 and 229 amino acids, have calculated molecular masses of 52 596 and 24 589 Da, respectively. TpβGRPc is 85.52% identical in sequence to TpβGRPa. TpβGRPb and TpβGRPd share the same deletion start site located at the conserved residue Pro 43, although TpβGRPd exhibits a much larger deletion of up to approximately 270 residues covering both the N‐ and C‐terminal regions. Affinity purification, associated with subsequent peptide sequencing, confirms the constitutive occurrence of TpβGRPa and TpβGRPc of similar size (approximately 65 kDa) in sixth‐instar larval haemolymph. These two βGRPs show clear binding affinities to curdlan, an insoluble β‐1,3‐glucan. A quantitative real‐time polymerase chain reaction analysis reveals the high‐level constitutive expression of TpβGRPc and TpβGRPd in the fat body of mid‐instar larvae, which are found to be susceptible to fungal pathogens in field investigations. Remarkable induction of both TpβGRPs occurs in response to haemocoelic challenge with entomopathogenic fungus Beauveria bassiana. The results of the present study suggest that TpβGRPs may contribute to the detection and control of fungal infections.

中文翻译：

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来自青藏高原鬼蛾 Thitarodes pui（鳞翅目，蛇形目）的新型 β-1,3-葡聚糖识别蛋白的表征

β-1,3-葡聚糖识别蛋白（βGRPs）在针对入侵病原体的先天免疫反应中作为模式识别受体发挥作用。在本研究中，我们从鬼蛾 Thitarodes pui（鳞翅目：蚧科）中获得了两种新型推定 βGRP 的全长 cDNA 克隆：TpβGRPc 和 TpβGRPd。系统发育分析显示了一个小的不同谱系，βGRP 进化枝 4，由 T. pui βGRP 组成，包括先前已鉴定的 TpβGRPa 和 TpβGRPb。TpβGRPc 和 TpβGRPd 由 488 和 229 个氨基酸组成，计算出的分子量分别为 52 596 和 24 589 Da。TpβGRPC 在序列上与 TpβGRPa 有 85.52% 的同一性。TpβGRPb 和 TpβGRPd 共享相同的缺失起始位点，位于保守残基 Pro 43，尽管 TpβGRPd 表现出更大的缺失，最多可达约 270 个残基，覆盖 N 端和 C 端区域。与随后的肽测序相关的亲和纯化证实了在六龄幼虫血淋巴中具有相似大小（约 65 kDa）的 TpβGRPa 和 TpβGRPc 的组成性发生。这两种 βGRP 显示出与 Curdlan（一种不溶性 β-1,3-葡聚糖）的明确结合亲和力。定量实时聚合酶链反应分析揭示了 TpβGRPc 和 TpβGRPd 在中龄幼虫脂肪体中的高水平组成型表达，在现场调查中发现它们对真菌病原体敏感。两种 TpβGRP 的显着诱导是响应于昆虫病原真菌白僵菌的血结肠攻击。