Scientia Agricultura Sinica ›› 2026, Vol. 59 ›› Issue (21): 4831-4840.doi: 10.3864/j.issn.0578-1752.2026.21.009

• PLANT PROTECTION • Previous Articles     Next Articles

Odorant-Binding Protein OBP4 Mediates Repellent Behavior of Bemisia tabaci to Host Plant Volatiles

JI Yuan(), SUN YuXiang, GONG Cheng, LIU HangWei(), LUO Chen   

  1. College of Plant Protection, Yangzhou University, Yangzhou 225000, Jiangsu
  • Received:2026-07-28 Accepted:2026-08-28 Online:2026-11-01 Published:2026-11-06
  • Contact: LIU HangWei

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Abstract:

【Background】The whitefly, Bemisia tabaci, is a highly polyphagous agricultural pest of global significance, exhibiting a distinct preference for specific host plants. Odorant-binding proteins (OBPs), which specifically bind to environmental odor molecules, play a pivotal role in insect behaviors such as host location, oviposition site selection, and predator avoidance. 【Objective】The purpose of this study is to deeply elucidate the molecular mechanism of OBP in the chemosensory and behavioral regulation of B. tabaci, reveal the physiological basis of its host adaptability, and to provide a theoretical basis for the development of new green prevention and control technologies based on olfactory interference. 【Method】Quantitative real-time PCR (qRT-PCR) was used to detect the expression profile of BtabOBP4 in different developmental stages (egg, 1st-4th instar nymphs, adult) and different adult tissues (head, thorax, abdomen) of B. tabaci. The full-length BtabOBP4 was cloned, and the recombinant BtabOBP4 protein was induced for heterologous expression and purified. Fluorescence competitive binding assays were conducted to screen the binding capacity of BtabOBP4 to plant volatiles and secondary metabolites. Y-tube olfactometer was used to determine the behavioral tropism of B. tabaci to candidate ligands. Feeding-mediated RNA interference (RNAi) technology was employed to silence the BtabOBP4 to verify its regulatory effect on chemosensory behavior in B. tabaci. 【Result】BtabOBP4 showed the highest expression level at the 3rd instar nymph stage of B. tabaci, and exhibited sexual dimorphism in the adult stage (male was approximately 4 times that of female), and was specifically enriched in the head. BtabOBP4 could specifically bind to myrcene, β-ionone, and α-pinene (the Ki values were 25.68, 43.95, and 33.65 μmol·L-1, respectively). Notably, 100 mg·L-1 myrcene, 100 mg·L-1 β-ionone, and 1 000 mg·L-1 α-pinene repelled B. tabaci by 33%, 43%, and 44%, respectively. After 24 h of dsRNA feeding, the silencing efficiency of BtabOBP4 reached 55% in females and 51% in males. The repellent responses of B. tabaci to myrcene and α-pinene were completely abolished, while the repellency toward β-ionone was significantly attenuated. 【Conclusion】BtabOBP4 is highly expressed in the 3rd instar nymphs and male adults of B. tabaci, with specific enrichment in the head tissue, suggesting its pivotal role in chemical communication. The BtabOBP4 protein exhibits specific binding affinity to myrcene, β-ionone, and α-pinene. Notably, silencing of BtabOBP4 via RNA interference significantly abolished or attenuated the repellent behavior of B. tabaci toward these compounds. These findings indicate that BtabOBP4 functions as a key olfactory receptor mediating host plant recognition and selection in B. tabaci, regulating repellent responses by perceiving specific plant volatiles.

Key words: Bemisia tabaci, odorant-binding protein (OBP), plant volatile, behavioral response, RNA interference (RNAi)

Table 1

Primer sequences"

引物Primer 引物序列Primer sequence (5′-3′) 引物用途Usage of primers
BtabOBP4-ORF-F gctgatatcggatccgaattcATGGCGCAAGACGCGAAG ORF全长扩增
ORF full-length amplification
BtabOBP4-ORF-R ctcgagtgcggccgcaagcttTCAAGCCTTGATGTAAAAGTTGTGC
BtabOBP4-qF TTGGTACATCCGAGCTTCCG 荧光定量PCR
qRT-PCR
BtabOBP4-qR CATCAGCCTCTTAGCCCCG
EF1α-qF TAGCCTTGTGCCAATTTCCG
EF1α-qR CCTTCAGCATTACCGTCC
BtOBP4-dsF TAATACGACTCACTATAGGGGACGCGAAGCAAAAAGTAGC RNAi
BtOBP4-dsR TAATACGACTCACTATAGGGGGGCCAAGGAACACTTCATA
dsEGFP-F TAATACGACTCACTATAGGGAGACAGTGCTTCAGCCGCTAC
dsEGFP-R TAATACGACTCACTATAGGGAGAGTTCACCTTGATGCCGTTC

Table 2

The compounds used in ligand binding assays"

化合物名称Compound name CAS号CAS number 化合物名称Compound name CAS号CAS number
植物挥发物Plant volatile α-石竹烯α-Caryophyllene 6753-98-6
1-辛醇1-Octanol 111-87-5 β-石竹烯β-Caryophyllene 87-44-5
2-庚酮2-Heptanone 110-43-0 香芹酚Carvacrol 499-75-2
α-蒎烯α-Pinene 80-56-8 2-甲基萘酚2-Methylnaphthol 135-19-3
α-萜品醇α-Terpineol 98-55-5 植物次生代谢物Plant secondary metabolite
β-紫罗兰酮β-Ionone 14901-07-6 儿茶素Catechin 154-23-4
莰烯Camphene 79-92-5 棉酚Gossypol 303-45-7
肉桂醇Cinnamyl alcohol 104-54-1 槲皮素Quercetin 117-39-5
丁酸乙酯Ethyl butyrate 105-54-4 奎宁Quinine 130-95-0
吲哚Indole 120-72-9 芦丁水合物Rutin hydrate 207671-50-9
芳樟醇Linalool 78-70-6 D-(-)-水杨苷D-(-)-Salicin 138-52-3
月桂烯Myrcene 123-35-3 单宁酸Tannic acid 1401-55-4
乙酸辛酯Octyl acetate 112-14-1 可可碱Theobromine 83-67-0
α-水芹烯α-Phellandrene 99-83-2 茶碱Theophylline 58-55-9
2-乙烯基-1-醇2-Ethen-1-ol 557-75-5 2-丁基-1-辛醇2-Butyl-1-octanol 3913-2-8
2-甲基萘2-Methylnaphthalene 91-57-6 2-乙基-1-癸醇2-Ethyl-1-decanol 25512-77-4

Fig. 1

The expression level of BtabOBP4 in different developmental stages and different tissues of adults B. tabaci Data are presented as mean±SD. Different lowercase letters on the column indicate significant differences among groups (P<0.05) by one-way ANOVA followed by Tukey’s multiple comparison test"

Fig. 2

Expression and purification of recombinant BtabOBP4 protein"

Fig. 3

Binding analysis of BtabOBP4 protein with 1-NPN"

Fig. 4

Competitive binding of candidate ligands with 1-NPN to recombinant BtabOBP4 protein"

Fig. 5

Behavioral responses of B. tabaci to myrcene, β- ionone, and α-pinene"

Fig. 6

In vivo functional validation of BtabOBP4"

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