韭菜迟眼蕈蚊气味结合蛋白OBP11的配体结合能力及关键氨基酸分析

doi:10.3864/j.issn.0578-1752.2026.18.005

中国农业科学 ›› 2026, Vol. 59 ›› Issue (18): 4007-4020.doi: 10.3864/j.issn.0578-1752.2026.18.005

• 专题：面向农业害虫精准防控的机制解析、技术优化与决策支撑 • 上一篇下一篇

韭菜迟眼蕈蚊气味结合蛋白OBP11的配体结合能力及关键氨基酸分析

王覃丽¹(), 谭顺阳¹, 夏根¹, 刘文庆², 杨玉婷¹(), 田丽霞²()

¹ 长江大学农学院/农业农村部长江中游作物绿色高效生产重点实验室（部省共建）/涝渍灾害与湿地农业湖北省重点实验室，湖北荆州 434025
² 北京市农林科学院植物保护研究所，北京 100097

收稿日期:2025-12-29 接受日期:2026-02-08 出版日期:2026-09-16 发布日期:2026-09-20
通信作者:
杨玉婷，E-mail：yangyuting198@163.com。
田丽霞，E-mail：tianlixia555@126.com。
联系方式: 王覃丽，E-mail：13477866904@163.com。
基金资助:
国家自然科学基金青年项目(32102202)

Ligand Binding Property and Key Amino Acid Analysis of Odorant- Binding Protein OBP11 in Bradysia odoriphaga

WANG QinLi¹(), TAN ShunYang¹, XIA Gen¹, LIU WenQing², YANG YuTing¹(), TIAN LiXia²()

¹ College of Agriculture, Yangtze University/Ministry of Agriculture and Rural Affairs Key Laboratory of Sustainable Crop Production in the Middle Reaches of the Yangtze River (Co-Construction by Ministry and Province)/Hubei Key Laboratory of Waterlogging Disaster and Agricultural Use of Wetland, Jingzhou 434025, Hubei
² Institute of Plant Protection, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097

Received:2025-12-29 Accepted:2026-02-08 Published:2026-09-16 Online:2026-09-20

摘要/Abstract

摘要：

【目的】明确韭菜迟眼蕈蚊（Bradysia odoriphaga）成虫触角高表达的气味结合蛋白BodoOBP11的分子特征，解析其在寄主定位中的嗅觉功能。【方法】利用生物信息学工具分析BodoOBP11的基本特征和进化关系；使用实时荧光定量PCR技术分析BodoOBP11在韭菜迟眼蕈蚊不同发育阶段和组织中的表达模式；通过大肠杆菌原核表达、镍柱亲和层析纯化和重组肠激酶酶切获得BodoOBP11蛋白；以1-NPN为荧光探针利用荧光竞争结合试验分析该蛋白与寄主植物韭菜的22种挥发性化合物和1种假定的性信息素化合物的结合能力；通过蛋白质结构预测、分子对接和定点突变揭示BodoOBP11蛋白与配体化合物的关键结合位点。【结果】BodoOBP11开放阅读框全长426 bp，编码141个氨基酸残基，在其氨基端有一段20个氨基酸残基组成的信号肽，且序列中具有Classic-C OBP的典型特征，即6个保守的半胱氨酸残基；其成熟蛋白预测分子量为13.53 kDa，等电点为7.71。实时荧光定量PCR结果表明，在不同发育阶段中，BodoOBP11在雄成虫中表达量最高，其次为雌成虫和蛹期。在不同组织中，BodoOBP11在雌、雄成虫的触角中均呈现显著高表达，且在雄虫触角中的表达量最高。荧光竞争结合试验显示，BodoOBP11仅与二丙基三硫醚、2-十三烷酮、二丙基二硫、2-十一烷酮、异硫氰酸苯乙酯及α-紫罗酮6种挥发物具有结合能力，其中与二丙基三硫醚结合能力最强。蛋白质结构预测与分子对接结果显示，BodoOBP11与上述6种配体的结合主要通过氢键与疏水相互作用，其中Thr74残基与2-十三烷酮、2-十一烷酮和α-紫罗酮形成氢键，而Val76、Ile107、Leu127、Met131、Phe138和Phe140等残基则主要介导疏水相互作用。定点突变后的荧光竞争结合试验显示，Ile107、Phe138和Phe140残基分别被突变成丙氨酸（Ala）后均会显著降低BodoOBP11与上述6种配体的结合能力，表明其为关键的结合位点。【结论】BodoOBP11对二丙基三硫醚、2-十三烷酮、二丙基二硫等6种韭菜挥发物表现出较强的结合亲和力，Ile107、Phe138和Phe140残基是其结合上述配体的关键残基。研究结果揭示了韭菜迟眼蕈蚊成虫利用BodoOBP11识别寄主植物挥发物的分子机制，可为基于嗅觉干扰的绿色防控技术研发提供依据。

关键词: 韭菜迟眼蕈蚊, 气味结合蛋白, 荧光竞争结合, 分子对接, 定点突变

Abstract:

【Objective】The objective of this study is to elucidate the molecular characteristics of the odorant-binding protein BodoOBP11, which is highly expressed in the antennae of adult Bradysia odoriphaga, and to investigate its olfactory function in host localization.【Method】Bioinformatics tools were used to analyze the basic characteristics and evolutionary relationships of BodoOBP11. The spatio-temporal expression patterns of BodoOBP11 across different developmental stages and tissues were then assessed using quantitative real-time PCR (qPCR). The BodoOBP11 protein was obtained through prokaryotic expression in E. coli, purification via nickel affinity chromatography, and cleavage using recombinant enterokinase. Using fluorescence competitive binding assays with the fluorescent probe 1-NPN, the binding ability of the protein to 22 volatile compounds from the host plant (Chinese chives) and one putative sex pheromone compound was subsequently evaluated. Finally, protein structure prediction, molecular docking, and site-directed mutagenesis were employed to identify the key binding sites between BodoOBP11 and ligand compounds.【Result】BodoOBP11 has an open reading frame of 426 bp, encoding 141 amino acid residues, including an N-terminal signal peptide of 20 amino acids. It exhibits the typical features of the Classic-C OBP, characterized by six conserved cysteine residues. The predicted molecular weight of the mature protein is 13.53 kDa, with an isoelectric point of 7.71. qPCR analysis revealed distinct expression patterns of BodoOBP11. Across developmental stages, its expression was highest in male adults, followed by female adults and pupae. In different tissues, BodoOBP11 showed significant high expression in the antennae of both sexes, with the highest level detected in male antennae. Fluorescence competitive binding assays demonstrated that BodoOBP11 binds to six volatiles: dipropyl trisulfide, 2-tridecanone, dipropyl disulfide, 2-undecanone, phenethyl isothiocyanate, and α-ionone. The strongest binding affinity was observed for dipropyl trisulfide. Protein structure prediction and molecular docking indicated that BodoOBP11 binds to these six ligands primarily via hydrogen bonding and hydrophobic interactions. Specifically, Thr74 forms hydrogen bonds with 2-tridecanone, 2-undecanone, and α-ionone, while residues Val76, Ile107, Leu127, Met131, Phe138, and Phe140 mainly mediate hydrophobic interactions. Site-directed mutagenesis followed by fluorescence competitive binding assays revealed that mutations of Ile107, Phe138, and Phe140 to alanine significantly reduced the binding affinity of BodoOBP11 to all six ligands, indicating their critical roles in binding.【Conclusion】BodoOBP11 exhibits strong binding affinity to six volatile compounds from Chinese chive, including dipropyl trisulfide, 2-tridecanone, and dipropyl disulfide, etc. Furthermore, residues Ile107, Phe138, and Phe140 were identified as crucial sites for binding these ligands. These findings preliminarily reveal its molecular mechanism in mediating host plant volatile recognition by adult B. odoriphaga using BodoOBP11, offering a theoretical foundation for developing green control technologies based on olfactory interference.

Key words: Bradysia odoriphaga, odorant-binding protein (OBP), fluorescence competitive binding, molecular docking, site-directed mutagenesis

王覃丽, 谭顺阳, 夏根, 刘文庆, 杨玉婷, 田丽霞. 韭菜迟眼蕈蚊气味结合蛋白OBP11的配体结合能力及关键氨基酸分析[J]. 中国农业科学, 2026, 59(18): 4007-4020.

WANG QinLi, TAN ShunYang, XIA Gen, LIU WenQing, YANG YuTing, TIAN LiXia. Ligand Binding Property and Key Amino Acid Analysis of Odorant- Binding Protein OBP11 in Bradysia odoriphaga[J]. Scientia Agricultura Sinica, 2026, 59(18): 4007-4020.

图/表 9

表1

图1

图2

图3

表2

重组蛋白BodoOBP11与不同挥发性化合物的结合力"

化合物名称 Compound name	分子式 Molecular formula	分子量 Molecular weight	CAS号 CAS number	BodoOBP11 (pH 7.4)
化合物名称 Compound name	分子式 Molecular formula	分子量 Molecular weight	CAS号 CAS number	IC₅₀(μmol·L^-1)	K_i (μmol·L^-1)
假定的性信息素Putative sex pheromone
正十七烷Heptadecane	C₁₇H₃₆	240.47	629-78-7	—	—
韭菜挥发物Chinese chives’ volatile
酮类Ketones
2-十一烷酮2-Undecanone	C₁₁H₂₂O	170.29	112-12-9	21.12±7.38	19.15±6.69
2-十三烷酮2-Tridecanone	C₁₃H₂₆O	198.34	593-08-8	15.58±2.95	14.13±2.67
α-紫罗酮α-Ionone	C₁₃H₂₀O	192.30	127-41-3	26.38±0.86	23.91±0.79
醇类Alcohols
3-甲基-2-丁烯-1-醇3-Methyl-2-buten-1-ol	C₅H₁₀O	86.13	556-82-1	—	—
芳樟醇Linalool	C₁₀H₁₈O	154.25	78-70-6	—	—
香茅醇Citronellol	C₁₀H₂₀O	156.27	106-22-9	—	—
醛类Aldehydes
苯甲醛Benzaldehyde	C₇H₆O	106.12	100-52-7	—	—
壬醛1-Nonanal	C₉H₁₈O	142.24	124-19-6	—	—
庚醛Heptaldehyde	C₇H₁₄O	114.19	111-71-7	—	—
挥发性硫类化合物Volatile sulfur compounds
二丙基二硫Propyl disulfide	C₆H₁₄S₂	150.31	629-19-6	18.33±2.80	16.62±2.54
二丙基三硫醚Dipropyl trisulfide	C₆H₁₄S₃	182.37	6028-61-1	5.25±0.47	4.76±0.43
二甲基三硫Dimethyl trisulfide	C₂H₆S₃	126.26	3658-80-8	—	—
二烯丙基二硫Diallyl disulfide	C₆H₁₀S₂	146.27	2179-57-9	—	—
甲基丙基二硫Methyl propyl disulfide	C₄H₁₀S₂	122.25	2179-60-4	—	—
不饱和烃类Unsaturated hydrocarbons
1-庚烯1-Heptene	C₇H₁₄	98.18	1161-13-3	—	—
石竹烯Caryophyllene	C₅₅H₂₄	204.36	87-44-5	—	—
β-蒎烯β-Pinene	C₁₀H₁₆	136.23	127-91-3	—	—
酯类Esters
异硫氰酸苯乙酯Phenylethyl isothiocyanate	C₉H₉NS	163.24	2257-09-2	21.67±1.66	19.64±1.51
3-（甲硫基）丙基异硫酸酯 3-(methyl thioyl) propyl isosulfate	C₅H₉NS₂	147.26	505-79-3	—	—
水杨酸甲酯Methyl salicylate	C₈H₈O₃	152.15	119-36-8	—	—
烯丙基硫氢酸酯Allyl thiocyanate	C₄H₅NS	99.15	764-49-8	—	—
月桂酸乙酯Ethyl dodecanoate	C₁₄H₂₈O₂	228.37	106-33-2	—	—

表2

图4

表3

图5

图6

参考文献 46

[1]	杨集昆, 张学敏. 韭菜蛆的鉴定迟眼蕈蚊属二新种(双翅目: 眼蕈蚊科)[J]. 北京农业大学学报, 1985, 11(2): 153-157.
	Yang J K, Zhang X M. Notes on the fragrant onion gnats with descriptions of two new species of Bradysia (Sciaridae: Diptera)[J]. Acta Agriculturae Universitatis Pekinensis, 1985, 11(2): 153-157. (in Chinese)
[2]	王覃丽, 王福, 华登科, 杨玉婷. 韭菜迟眼蕈蚊气味受体BodoOR71和BodoOR72基因时空表达分析[J]. 中国蔬菜, 2024(6): 55-64.
	Wang Q L, Wang F, Hua D K, Yang Y T. Spatiotemporal expression analysis of odorant receptor genes of BodoOR71 and BodoOR72 in Bradysia odoriphaga[J]. China Vegetables, 2024(6): 55-64. (in Chinese)
[3]	吴青君, 于毅, 谷希树, 刘峰, 宋敦伦, 魏国树, 贺敏, 刘长仲, 许国庆, 张友军. 韭菜根蛆的发生危害及综合防治技术研究——公益性行业(农业)科研专项“作物根蛆类害虫综合防治技术研究与示范”进展[J]. 应用昆虫学报, 2016, 53(6): 1165-1173.
	Wu Q J, Yu Y, Gu X S, Liu F, Song D L, Wei G S, He M, Liu C Z, Xu G Q, Zhang Y J. The occurrence of, and damage caused by, root maggots on Chinese chives and integrated management techniques to control these pests[J]. Chinese Journal of Applied Entomology, 2016, 53(6): 1165-1173. (in Chinese)
[4]	杨玉婷, 梁玲, 史彩华, 谢文, 张友军. 不同温度下韭菜迟眼蕈蚊年龄-阶段两性实验种群生命表的构建[J]. 植物保护, 2019, 45(1): 51-57.
	Yang Y T, Liang L, Shi C H, Xie W, Zhang Y J. Age-stage, two-sex life table of laboratory populations of Bradysia odoriphaga Yang et Zhang under different temperatures[J]. Plant Protection, 2019, 45(1): 51-57. (in Chinese)
[5]	杨玉婷, 史彩华, 程佳旭, 张友军. 不同寄主植物对韭菜迟眼蕈蚊生长发育、繁殖和保护酶活力的影响[J]. 植物保护, 2017, 43(5): 119-123.
	Yang Y T, Shi C H, Cheng J X, Zhang Y J. Effects of feeding on different host plants on development, reproduction and protective enzymes of Bradysia odoriphaga[J]. Plant Protection, 2017, 43(5): 119-123. (in Chinese)
[6]	Li X, Zhang X G, Xiao C, Gao Y L, Dong W X. Behavioral responses of potato tuber moth (Phthorimaea operculella) to tobacco plant volatiles[J]. Journal of Integrative Agriculture, 2020, 19(2): 325-332. doi: 10.1016/S2095-3119(19)62663-8
[7]	Zhang L, Jiang H Q, Wu F, Wen P, Qing J, Song X M, Li H L. Eastern honeybee Apis cerana sense cold flowering plants by increasing the static binding affinity of odorant-binding protein to cold floral volatiles from loquats[J]. International Journal of Biological Macromolecules, 2023, 232: 123227. doi: 10.1016/j.ijbiomac.2023.123227
[8]	Li D X, Li C B, Liu D G. Analyses of structural dynamics revealed flexible binding mechanism for the Agrilus mali odorant binding protein 8 towards plant volatiles[J]. Pest Management Science, 2021, 77(4): 1642-1653. doi: 10.1002/ps.v77.4
[9]	Yang Y T, Wang F, Wang Q L, Tan S Y, Zhou Z X, Su Q, Hua D K, Tian L X. Functional characterization of BodoCSP3 in odorant recognition and host-seeking behavior of the insect pest Bradysia odoriphaga (Diptera: Sciaridae)[J]. Pest Management Science, 2026, 82(3): 2206-2215. doi: 10.1002/ps.v82.3
[10]	Jing T T, Qian X N, Du W K, Gao T, Li D F, Guo D Y, He F, Yu G M, Li S P, Schwab W, Wan X C, Sun X L, Song C K. Herbivore-induced volatiles influence moth preference by increasing the β-ocimene emission of neighbouring tea plants[J]. Plant, Cell and Environment, 2021, 44(11): 3667-3680. doi: 10.1111/pce.v44.11
[11]	Britch S C, Dame D A, Meisch M V, Kline D L, Walker T W, Allan S A, Urban J, Aldridge R L, Linthicum K J. Spatial repellents protect small perimeters from riceland mosquitoes in a warm-humid environment[J]. Journal of the American Mosquito Control Association, 2021, 37(1): 41-45. doi: 10.2987/20-6974.1 pmid: 33857316
[12]	Yang Y T, Su Q, Shi L L, Chen G, Zeng Y, Shi C H, Zhang Y J. Electrophysiological and behavioral responses of Bradysia odoriphaga (Diptera: Sciaridae) to volatiles from its host plant, Chinese chives (Allium tuberosum Rottler ex Spreng)[J]. Journal of Economic Entomology, 2019, 112(4): 1638-1644. doi: 10.1093/jee/toz057
[13]	Pelosi P, Zhou J J, Ban L P, Calvello M. Soluble proteins in insect chemical communication[J]. Cellular and Molecular Life Sciences, 2006, 63(14): 1658-1676. doi: 10.1007/s00018-005-5607-0 pmid: 16786224
[14]	Zhou J J. Odorant-binding proteins in insects[M]// Vitamins and Hormones. Amsterdam: Elsevier, 2010, 83: 241-272.
[15]	Leal W S, Nikonova L, Peng G. Disulfide structure of the pheromone binding protein from the silkworm moth, Bombyx mori[J]. FEBS Letters, 1999, 464(2): 85-90. doi: 10.1016/S0014-5793(99)01683-X
[16]	Brito N F, Moreira M F, Melo A C A. A look inside odorant-binding proteins in insect chemoreception[J]. Journal of Insect Physiology, 2016, 95: 51-65. doi: S0022-1910(16)30108-1 pmid: 27639942
[17]	Sun L, Wang Q, Wang Q, Dong K, Xiao Y, Zhang Y J. Identiﬁcation and characterization of odorant binding proteins in the forelegs of Adelphocoris lineolatus (Goeze)[J]. Frontiers in Physiology, 2017, 8: 735. doi: 10.3389/fphys.2017.00735
[18]	Liu X Q, Jiang H B, Fan J Y, Liu T Y, Meng L W, Liu Y, Yu H Z, Dou W, Wang J J. An odorant-binding protein of Asian citrus psyllid, Diaphorina citri, participates in the response of host plant volatiles[J]. Pest Management Science, 2021, 77(7): 3068-3079. doi: 10.1002/ps.v77.7
[19]	Wang Q, Liu J T, Zhang Y J, Chen J L, Li X C, Liang P, Gao X W, Zhou J J, Gu S H. Coordinative mediation of the response to alarm pheromones by three odorant binding proteins in the green peach aphid Myzus persicae[J]. Insect Biochemistry and Molecular Biology, 2021, 130: 103528. doi: 10.1016/j.ibmb.2021.103528
[20]	Li E T, Wu H J, Qin J H, Luo J, Li K B, Cao Y Z, Zhang S, Peng Y, Yin J. Involvement of Holotrichia parallela odorant binding protein 3 in the localization of oviposition sites[J]. International Journal of Biological Macromolecules, 2023, 242: 124744. doi: 10.1016/j.ijbiomac.2023.124744
[21]	Zhao Y H, Ding J F, Zhang Z Q, Liu F, Zhou C G, Mu W. Sex- and tissue-specific expression profiles of odorant binding protein and chemosensory protein genes in Bradysia odoriphaga (Diptera: Sciaridae)[J]. Frontiers in Physiology, 2018, 9: 107. doi: 10.3389/fphys.2018.00107
[22]	Tang B W, Tai S L, Dai W, Zhang C N. Expression and functional analysis of two odorant-binding proteins from Bradysia odoriphaga (Diptera: Sciaridae)[J]. Journal of Agricultural and Food Chemistry, 2019, 67(13): 3565-3574. doi: 10.1021/acs.jafc.9b00568
[23]	Yang Y T, Hua D K, Shi C H, Xie W, Zhang Y J. Molecular and binding characteristics of OBP5 of Bradysia odoriphaga (Diptera: Sciaridae)[J]. Journal of Economic Entomology, 2021, 114(4): 1509-1516. doi: 10.1093/jee/toab095
[24]	Yang Y T, Tan S Y, Wang Q L, Wang F, Zhang Y J. Key amino acids in odorant-binding protein OBP7 enable Bradysia odoriphaga to recognize host plant volatiles[J]. International Journal of Biological Macromolecules, 2025, 284(2): 138179. doi: 10.1016/j.ijbiomac.2024.138179
[25]	Yang Y T, Luo L, Tian L X, Zhao C W, Niu H L, Hu Y F, Shi C H, Xie W, Zhang Y J. Function and characterization analysis of BodoOBP8 from Bradysia odoriphaga (Diptera: Sciaridae) in the recognition of plant volatiles and sex pheromones[J]. Insects, 2021, 12(10): 879. doi: 10.3390/insects12100879
[26]	Zhu J Q, Wang F, Zhang Y J, Yang Y T, Hua D K. Odorant binding protein 10 from Bradysia odoriphaga (Diptera: Sciaridae) binds volatile host plant compounds[J]. Journal of Insect Science, 2023, 23(1): 7.
[27]	王鸿梅, 冯静. 韭菜挥发油中化学成分的研究[J]. 天津医科大学学报, 2002, 8(2): 191-192.
	Wang H M, Feng J. Identification on chemical components of volatile oil of chive by GC/MS[J]. Journal of Tianjin Medical University, 2002, 8(2): 191-192. (in Chinese)
[28]	Li H J, He X K, Zeng A J, Liu Y J, Jiang S R. Bradysia odoriphaga copulatory behavior and evidence of a female sex pheromone[J]. Journal of Agricultural and Urban Entomology, 2007, 24(1): 27-34. doi: 10.3954/1523-5475-24.1.27
[29]	Trifinopoulos J, Nguyen L T, von Haeseler A, Minh B Q. W-IQ-TREE: A fast online phylogenetic tool for maximum likelihood analysis[J]. Nucleic Acids Research, 2016, 44(1): 232-235. doi: 10.1093/nar/gkv997
[30]	Pfaffl M W. A new mathematical model for relative quantification in real-time RT-PCR[J]. Nucleic Acids Research, 2001, 29(9): e45. doi: 10.1093/nar/29.9.e45 pmid: 11328886
[31]	Silva D, Ceballos R, Arismendi N, Dalmon A, Vargas M. Variant A of the deformed wings virus alters the olfactory sensitivity and the expression of odorant binding proteins on antennas of Apis mellifera[J]. Insects, 2021, 12(10): 895. doi: 10.3390/insects12100895
[32]	Li X H, Hu F J, Li R F, Peng D H, Gao P, Rao F Q, Li Y H, Liu D G. The pleiotropic odorant binding protein CaspOBP12 involved in perception of Ceutorhynchus asper for plant volatiles and pesticides[J]. Pesticide Biochemistry and Physiology, 2025, 214: 106578. doi: 10.1016/j.pestbp.2025.106578
[33]	Chen Y W, Yang H H, Gu N, Li J Q, Zhu X Y, Zhang Y N. Identification of attractants for adult Spodoptera litura based on the interaction between odorant-binding protein 34 and host volatiles[J]. Pesticide Biochemistry and Physiology, 2024, 203: 106005. doi: 10.1016/j.pestbp.2024.106005
[34]	Wang Q K, Liu G T, Yan L P, Xu W T, Hilton D J, Liu X H, Pei W Y, Li X Y, Wu J B, Zhao H F, Zhang D, Elgar M A. Short-term particulate matter contamination severely compromises insect antennal olfactory perception[J]. Nature Communications, 2023, 14: 4112. doi: 10.1038/s41467-023-39469-3 pmid: 37433781
[35]	Liu H, Zhao X F, Fu L, Han Y Y, Chen J, Lu Y Y. BdorOBP2 plays an indispensable role in the perception of methyl eugenol by mature males of Bactrocera dorsalis (Hendel)[J]. Scientific Reports, 2017, 7: 15894. doi: 10.1038/s41598-017-15893-6
[36]	Wang H, Ma Y F, Wang M M, Chen G L, Dewer Y, He M, Zhang F, Yang Y F, Liu J F, He P. Expression, affinity, and functional characterization of the specific binding of two putative pheromone- binding proteins in the omnivorous German cockroach Blattella germanica[J]. Journal of Agricultural and Food Chemistry, 2020, 68(47): 13573-13583. doi: 10.1021/acs.jafc.0c02456
[37]	赵云贺. 韭菜迟眼蕈蚊化学感受基因的鉴定、克隆及功能研究[D]. 泰安: 山东农业大学, 2019.
	Zhao Y H. Identification, cloning and functional characterization of chemosensory genes in Bradysia odoriphaga (Diptera: Sciaridae)[D]. Taian: Shandong Agricultural University, 2019. (in Chinese)
[38]	Obata T, Koh H S, Kim M, Fukami H. Constituents of planthopper attractant in rice plant[J]. Applied Entomology and Zoology, 1983, 18(2): 161-169. doi: 10.1303/aez.18.161
[39]	Wen M, Li E T, Li J Q, Chen Q, Zhou H F, Zhang S, Li K B, Ren B Z, Wang Y L, Yin J. Molecular characterization and key binding sites of sex pheromone-binding proteins from the meadow moth, Loxostege sticticalis[J]. Journal of Agricultural and Food Chemistry, 2019, 67(46): 12685-12695. doi: 10.1021/acs.jafc.9b03235
[40]	Hao E H, Li Y N, Guo B, Yang X, Lu P F, Qiao H L. Key residues affecting binding affinity of Sirex noctilio Fabricius odorant-binding protein (SnocOBP9) to aggregation pheromone[J]. International Journal of Molecular Sciences, 2022, 23(15): 8456. doi: 10.3390/ijms23158456
[41]	Li H L, Song X M, Wu F, Qiu Y L, Fu X B, Zhang L Y, Tan J. Chemical structure of semiochemicals and key binding sites together determine the olfactory functional modes of odorant-binding protein 2 in eastern honey bee, Apis cerana[J]. International Journal of Biological Macromolecules, 2020, 145: 876-884. doi: 10.1016/j.ijbiomac.2019.11.189
[42]	del Mármol J, Yedlin M A, Ruta V. The structural basis of odorant recognition in insect olfactory receptors[J]. Nature, 2021, 597: 126-131. doi: 10.1038/s41586-021-03794-8
[43]	Xiang D K, Abdelnabby H, Wang M Q. Predicted structure of odorant-binding protein 12 from Monochamus alternatus (Hope) suggests a mechanism of flexible odorant-binding[J]. International Journal of Biological Macromolecules, 2023, 243: 125152. doi: 10.1016/j.ijbiomac.2023.125152
[44]	付淑慧, 张鹤, 赵婉清, 梁志刚. 小菜蛾气味结合蛋白PxylOBP33与其相关信息化学物质的分子对接[J]. 环境昆虫学报, 2023, 45(5): 1291-1305.
	Fu S H, Zhang H, Zhao W Q, Liang Z G. Molecular docking of Plutella xylostella odorant binding protein PxylOBP33 with its related semiochemicals[J]. Environmental Entomology, 2023, 45(5): 1291-1305. (in Chinese)
[45]	Liu Z, Tang D, Lai Y, Gao S, Han H, Zhou Y. Prokaryotic expression and binding characteristics of odor-binding protein GqinOBP10 in Gynaephora qinghaiensis[J]. International Journal of Molecular Sciences, 2025, 26(21): 10502. doi: 10.3390/ijms262110502
[46]	Yang H H, Li S P, Yin M Z, Zhu X Y, Li J B, Zhang Y N, Li X M. Functional differentiation of two general odorant-binding proteins to sex pheromones in Spodoptera frugiperda[J]. Pesticide Biochemistry and Physiology, 2023, 191: 105348. doi: 10.1016/j.pestbp.2023.105348

引物名称Primer name	引物序列Primer sequence
用于克隆 For cloning	F: CGCGGATCCGGTGTAACGATGGAGCAAAT
用于克隆 For cloning	R: GGGAAGCTTTTAAGCAAACGTGAACTTTTTAT
用于实时荧光定量分析 For qPCR analysis	F: AACTATGAGGCATCTCTGAA
用于实时荧光定量分析 For qPCR analysis	R: TTACTCCATTAGCAGCATCT
内参基因 Reference gene	RPS15-F: ATCGTGGCGTCGATTTGGAT
	RPS15-R: CTCATTTGGTGGGGCTTCCT
	RPL18-F: TTCCCTTATCCATCAAACATT
	RPL18-R: CTTAGAATAACTTCAGGTAATGC
用于原核表达 For heterologous expression	F: CGCGGATCCGGTGTAACGATGGAGCAAAT
用于原核表达 For heterologous expression	R: GGGAAGCTTTTAAGCAAACGTGAACTTTTTAT
基因定点突变 Site-directed mutant	T74A-F: GCCGGAGTCATGAAGAAAAACAAACT
	T74A-R: CATTTCCAACACACATTGCATGT
	V76A-F: GCCATGAAGAAAAACAAACT
	V76A-R: TCCTGTCATTTCCAACACACAT
	I107A-F: GCCACGCAGTGCAAAGATGC
	I107A-R: ACCATTTTTATTTTCTTCCACAATGTCGT
	L127A-F: GCCCTTCCTAATAAGAT
	L127A-R: TATGTATGCTGCTTCGCAGGCAT
	M131A-F: GCCTACAAAAACAATAAAAAGTTCACGT
	M131A-R: GCATTGAACTAATATGTATGCTGCT
	P138A-F: CGCGGATCCGGTGTAACGATGGAGCAAAT
	P138A-R: GGGAAGCTTTTAAGCAAACGTGGCCTTTT
	P140A-F: CGCGGATCCGGTGTAACGATGGAGCAAAT
	P140A-R: GGGAAGCTTTTAAGCGGCCGTGAACT

配体名称 Ligand name	结合能 Binding energy (kJ·mol^-1)	氢键 Conventional hydrogen bond	疏水相互作用 Hydrophobic interaction
二丙基二硫Propyl disulfide	-15.23	—	Lys81, Leu82, Ala114, Leu127
二丙基三硫醚Dipropyl trisulfide	-16.61	—	Val76, Leu82, Ile107, Phe138, Phe140
2-十三烷酮2-Tridecanone	-21.67	Thr74	Leu71, Val76, Leu82, Gln90, Ile107, Leu127, Met131, Phe138, Phe140
2-十一烷酮2-Undecanone	-20.71	Thr74	Leu71, Val76, Gln90, Ile107, Leu127, Val128, Met131, Phe138, Phe140
α-紫罗酮α-Ionone	-26.02	Thr74	Val76, Gln90, Leu94, Ile107, Met131, Phe138, Phe140
异硫氰酸苯乙酯Phenylethyl isothiocyanate	-21.46	—	Leu71, Ile107, Leu127, Val128, Met131, Phe138, Phe140

韭菜迟眼蕈蚊气味结合蛋白OBP11的配体结合能力及关键氨基酸分析

Ligand Binding Property and Key Amino Acid Analysis of Odorant- Binding Protein OBP11 in Bradysia odoriphaga

RichHTML

PDF

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 9

参考文献 46

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	姚宏, 施寿荣, 赵茹茜. 基于网络药理学和分子对接探究绿原酸缓解鸡肠道炎症的作用机制[J]. 中国农业科学, 2025, 58(3): 600-616.
[2]	王梦云, 邓力元, 余洋, 杨宇衡, 方安菲, 田斌年, 王静, 毕朝位. 稻瘟病菌琥珀酸脱氢酶不同位点氨基酸替换对苯并烯氟菌唑敏感性的影响[J]. 中国农业科学, 2025, 58(24): 5175-5189.
[3]	刘文龙, 常译方, 李文宇, 孙丽娟, 郑长英. 蛹期干旱胁迫对韭菜迟眼蕈蚊交配及虱螨脲敏感性的影响[J]. 中国农业科学, 2025, 58(14): 2793-2804.
[4]	刘卓琳, 刘红云. 基于网络药理学和分子对接探究芹菜素缓解奶牛热应激及低氧应激的潜力与机制[J]. 中国农业科学, 2024, 57(5): 1010-1022.
[5]	汪志雄, 许冬, 田晓丽, 万鹏, 夏根, 宋旭荣, 王福莲, 桂连友, 张国辉. 柑橘大实蝇BminMinusOBP1和BminPlusOBP1的克隆、原核表达及其配体结合特性[J]. 中国农业科学, 2024, 57(24): 4894-4906.
[6]	徐重新, 金嘉凤, 孙晓明, 沈成, 张霄, 陈澄宇, 刘贤金, 刘媛. 基于Bt毒素的杀虫蛋白理性设计与创新应用策略[J]. 中国农业科学, 2024, 57(1): 96-125.
[7]	杨岭, 田晓丽, 桂连友, 王福莲, 张国辉. 柑橘大实蝇气味结合蛋白BminOBP6与其气味配体的互作机制[J]. 中国农业科学, 2023, 56(7): 1311-1321.
[8]	年和粉, 张钰析, 李伯辽, 陈秀琳, 罗坤, 李广伟. 李小食心虫GfunOBP2的原核表达及气味配体结合特性[J]. 中国农业科学, 2023, 56(12): 2302-2316.
[9]	李桂香,李秀环,郝新昌,李智文,刘峰,刘西莉. 山东省多主棒孢对三种常用杀菌剂的敏感性监测及对氟吡菌酰胺的抗性[J]. 中国农业科学, 2022, 55(7): 1359-1370.
[10]	赵慧婷,彭竹,姜玉锁,赵淑果,黄丽,杜亚丽,郭丽娜. 中华蜜蜂气味结合蛋白AcerOBP7的表达及结合特性[J]. 中国农业科学, 2022, 55(3): 613-624.
[11]	刘孝贺,仇贵生,佟兆国,张怀江,闫文涛,岳强,孙丽娜. 桃小食心虫化学感受蛋白CSP16配体结合特性[J]. 中国农业科学, 2021, 54(5): 945-958.
[12]	秦健辉,李金桥,赵旭,李克斌,曹雅忠,尹姣. 铜绿丽金龟气味结合蛋白AcorOBP11的表达纯化及功能分析[J]. 中国农业科学, 2021, 54(14): 3017-3028.
[13]	李祖任,罗丁峰,柏浩东,徐晶晶,韩进财,徐强,王若仲,柏连阳. 小飞蓬捕光叶绿素结合蛋白基因CcLhca-J9克隆及表达分析[J]. 中国农业科学, 2021, 54(1): 86-94.
[14]	李玲,谭瑶,周晓榕,庞保平. 沙葱萤叶甲气味结合蛋白GdauOBP20的基因克隆、原核表达及其结合特性[J]. 中国农业科学, 2019, 52(20): 3705-3712.
[15]	李都, 牛长缨, 李峰奇, 罗晨. 斑翅果蝇气味结合蛋白OBP56h与小分子化合物的结合特征[J]. 中国农业科学, 2019, 52(15): 2616-2623.