| Simulium maculatum is one of the most important medical insects in the world.It not only sucks human and animal blood,but also spreads virus and pathogenic bacteria,which seriously affects the agricultural and animal husbandry production and people's life in Altay,Xinjiang.At present,biological pesticides and physical control are used to control black flies at home and abroad,but the effect is limited.As people pay more and more attention to environmental protection,insect resistance and insecticide pollution,it is urgent to develop a green and efficient prevention and control method.In this paper,Simulium maculatum were selected as the research object.The functional green plant source compounds obtained by behavioral screening and the functional compounds specifically combined with odorant binding protein(OBP)of Simulium maculatum were linked.Through field test,effective attractants and repellents were found,which provided theoretical guidance for green control of Simulium maculatum and developed the control agents in trap products.In addition,based on the macrogenomic data of Simulium maculatum,the information of microbial species composition was screened to lay a foundation for its microbial control.The main results are as follows:(1)72 species of volatiles from green plants were screened from the habitats of Simulium maculatum.Through the Y-tube double-selection experiment,the functional green compounds were screened out.Among them,there were 7 kinds of compounds which had significant attraction to the Simulium maculatum,They are: Dibutyl phthalate,(-)Linalool,Frankincense Essential Oil,NENU-2,Grapefruit Essential Oil,Sweet Fennel Essential Oil and Tea Tree Essential Oil.There are 10 compounds which have significant avoidance effect on the Simulium maculatum.They are:Orthoxylene,Methyl salicylate,Dodecane,Ethyl linoleate,Methyl jasmonate,NENU-1,Thyme Essential Oil,Lavender Essential Oil,Citrus Essential Oil,Eucalyptus Rosemary Essential Oil.(2)A total of 17 OBP sequences,including one GOBP sequence,were screened from the antennae transcriptome data.The in vitro pure protein SmacOBP1 was obtained by E.coli prokaryotic expression system and purification system.The results of fluorescent competition binding experiments showed that the binding affinity of SmacOBP1 is more specific,only for NENU-2,the binding affinity was high,which provided the target compound for field behavioral test.(3)Field behavioral tests found that NENU-3 has the most obvious trapping effect,followed by NENU-2,two kinds of essential oils can be used as the trapping core of a new type of trap,and samples of the Simulium maculatum trap were developed.At the same time,it is found that the avoidance rate of NENU-1 is 73.8%,which can be developed as a avoidant.(4)A total of 367 Familys,694 Genus and 1320 Species were screened from metagenomic data,of which Debaryomyces prosopidis had the highest proportion,Bacillus cereus,Escherichia coli and Clostridium botulinum accounts for a relatively high proportion.Metagenomic data provides data for the study on the species and richness of the intestinal microflora,and studies and analyzes the functions of some reported microorganisms,which lays a foundation for the research of microbial control in Simulium maculatum.In summary,through the combination of behavioral experiments and fluorescence competition experiments,this article screened out the attractant NENU-3and NENU-2 with obvious trapping effect,and the repellent NENU-1 with good avoidance effect,and successfully developed the sample of the Simulium maculatum trap,whicn opened up a new way for the green chemical control of the Simulium maculatum,and it has important guiding significance in the prevention and control of the olfactory gene of the Simulium maculatum.On the other hand,the screened microbial species data provides a theoretical research foundation for the biological control of Simulium maculatum. |
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