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Simple Summary: The tea green leafhopper, Empoasca flavescens Fabrecius, is the most notorious pest in Chinese tea plantations, predominantly controlled through pesticide applications for decades. Recognizing the pressing need for novel, effective, and eco-friendly strategies to manage leafhoppers, this study aims to contribute to the reduction in pesticide usage in the tea industry. Salivary protein research is pivotal for gaining insights into effectors or elicitors, facilitating the exploration of novel regulatory targets and pathways in tea green leafhoppers. In this study, saliva from adult tea green leafhoppers was collected using a self-made collection device equipped with two layers of Parafilm stretched over a sucrose diet. Following freeze drying and filter-aided sample preparation (FASP), liquid chromatography–tandem mass spectrometry (LC-MS/MS) was employed to analyze the saliva proteins. Bioinformatics methodologies were then leveraged to predict the secretory proteins. The results suggest that the interactions between leafhoppers and tea plants, particularly in terms of feeding and defense responses, may be mediated by elicitors or effectors, such as mucin-like protein (EfMucin), vitellogenin (EfVg), odorant-binding protein (EfOBP), and others. The conclusive outcomes of this study provide new insights into the coevolutionary dynamics between tea plants and leafhoppers, offering novel pathways for the development of advanced leafhopper control technologies. Saliva plays a crucial role in shaping the compatibility of piercing–sucking insects with their host plants. Understanding the complex composition of leafhopper saliva is important for developing effective and eco-friendly control strategies for the tea green leafhopper, Empoasca flavescens Fabrecius, a major piercing–sucking pest in Chinese tea plantations. This study explored the saliva proteins of tea green leafhopper adults using a custom collection device, consisting of two layers of Parafilm stretched over a sucrose diet. A total of 152 proteins were identified using liquid chromatography–tandem mass spectrometry (LC-MS/MS) following the filter-aided sample preparation (FASP). These proteins were categorized into six groups based on their functions, including enzymes, transport proteins, regulatory proteins, cell structure proteins, other proteins, and unknown proteins. Bioinformatics analyses predicted 16 secreted proteins, which were successfully cloned and transcriptionally analyzed across various tissues and developmental stages. Genes encoding putative salivary secretory proteins, including Efmucin1, EfOBP1, EfOBP2, EfOBP3, Efmucin2, low-density lipoprotein receptor-related protein (EfLRP), EFVg1, and EFVg2, exhibited high expressions in salivary gland (SG) tissues and feeding-associated expressions at different developmental stages. These findings shed light on the potential elicitors or effectors mediating the leafhopper feeding and defense responses in tea plants, providing insights into the coevolution of tea plants and leafhoppers. The study's conclusions open avenues for the development of innovative leafhopper control technologies that reduce the reliance on pesticides in the tea industry. [ABSTRACT FROM AUTHOR]

BACKGROUND The tea green leafhopper, Empoasca flavescens is the most important pest in Chinese tea plantations. For decades its control has been executed almost exclusively through pesticide applications. A semiochemical‐based 'push–pull' strategy was tested on the leafhopper in the study. RESULTS: The odors released from Tagetes erecta and Flemingia macrophylla significantly repelled and attracted leafhoppers, respectively. These volatile compounds (46 from T. erecta and 53 F. macrophylla) were identified and quantified via gas chromatography–mass spectometry (GC–MS) analysis. Y‐tube olfactometer assays indicated that thymol anisole, thymol and camphor had significant repellent effects on the leafhoppers, resulting in a ternary repellent blend at a 4:3:13 ratio. Cis‐3‐hexen‐1‐ol, cis‐3‐hexenyl acetate, nonanal and α‐farnesene were significantly attractive to the leafhoppers, making an attractant blend at a 17:4:1:1 ratio. In the field, the push–pull strategy with the repellent dispensers placed within the tea bushes and the attractant‐baited sticky traps hung 15 cm above the tea plants showed a significant control efficacy, reaching 69% and 55% at two and 14 days post‐treatment, respectively, similar to those in the insecticide control plots. Additionally, the leafhopper density in the push–pull intercropping plot was 63.2 leafhoppers/100 tea shoots/visit, much lower than those in the pull intercropping plot and nonintercropping plot. CONCLUSION: Application of the push–pull strategy using both synthetic repellent and attractant, or intercropping T. erecta and F. macrophylla with tea plants, can effectively reduce the leafhopper population. This approach might have great potential as an environmentally safe control strategy against the leafhopper. © 2022 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]