State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Yuanmingyuan West Road, Beijing 100193, China.
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Yuanmingyuan West Road, Beijing 100193, China; Hubei Key Laboratory of Regional Development and Environmental Response, Faculty of Resources and Environmental Science, Hubei University, Wuhan, China.
Int J Biol Macromol. 2023 Jul 1;242(Pt 1):124744. doi: 10.1016/j.ijbiomac.2023.124744. Epub 2023 May 5.
Organic fertilizers-derived volatiles attract Holotrichia parallela during oviposition. However, the mechanisms underlying the perception of oviposition cues in H. parallela remain unclear. Here, H. parallela odorant-binding protein 3 (HparOBP3) was identified as a key OBP. Bioinformatics analysis showed that HparOBP3 clustered together with Holotrichia oblita OBP8. HparOBP3 was mainly expressed in the antennae of both sexes. Recombinant HparOBP3 exhibited distinct binding affinities towards 22 compounds released by organic fertilizers. After 48 h of RNA interference (RNAi), the expression of HparOBP3 in male and female antennae was decreased by 90.77 % and 82.30 %, respectively. In addition, silencing of HparOBP3 significantly reduced the electrophysiological responses and tropism of males to cis-3-hexen-1-ol, 1-hexanol, and (Z)-β-ocimene as well as females to cis-3-hexen-1-ol, 1-hexanol, benzaldehyde, and (Z)-β-ocimene. Molecular docking indicated that hydrophobic residues Leu-83, Leu-87, Phe-108, and Ile-120 of HparOBP3 were important amino acids for interacting with ligands. Mutation of the key residue, Leu-83, significantly diminished the binding ability of HparOBP3. Furthermore, acrylic plastic arena bioassays showed that the attraction and oviposition indexes of organic fertilizers to H. parallela were reduced by 55.78 % and 60.11 %, respectively, after silencing HparOBP3. These results suggest that HparOBP3 is essential in mediating the oviposition behavior of H. parallela.
有机肥衍生挥发物吸引暗黑鳃金龟产卵。然而，暗黑鳃金龟产卵线索感知的机制尚不清楚。在这里，鉴定出暗黑鳃金龟气味结合蛋白 3（HparOBP3）作为一种关键的 OBPs。生物信息学分析表明，HparOBP3 与直翅目昆虫丽金龟属 OBP8 聚类在一起。HparOBP3 主要在两性触角中表达。重组 HparOBP3 对有机肥释放的 22 种化合物表现出明显的结合亲和力。经 48 h RNA 干扰（RNAi）后，雄性和雌性触角中 HparOBP3 的表达分别降低了 90.77%和 82.30%。此外，沉默 HparOBP3 显著降低了雄性对顺-3-己烯-1-醇、1-己醇和（Z）-β-罗勒烯以及雌性对顺-3-己烯-1-醇、1-己醇、苯甲醛和（Z）-β-罗勒烯的电生理反应和趋向性。分子对接表明 HparOBP3 的疏水残基 Leu-83、Leu-87、Phe-108 和 Ile-120 是与配体相互作用的重要氨基酸。关键残基亮氨酸 83 的突变显著降低了 HparOBP3 的结合能力。此外，丙烯酸塑料竞技场生物测定表明，沉默 HparOBP3 后，有机肥对暗黑鳃金龟的吸引力和产卵指数分别降低了 55.78%和 60.11%。这些结果表明，HparOBP3 在介导暗黑鳃金龟的产卵行为中是必不可少的。
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