| Tick is a kind of parasitic arthropod that sucks blood exclusively.It mainly parasitizes on the body surface of mammals,birds,reptiles and amphibians.It is considered to be one of the first terrestrial arachnids to develop the ability of sucking blood.Haemaphysalis longicornis belongs to Ixodidae and Haemaphysalis.This species is one of the main tick species in China.It is the carrier of a variety of pathogens and can spread a variety of diseases.It is very harmful to human,livestock and wild animals.Salivary gland,as one of the most important organs of ticks,plays an important role in promoting blood sucking,regulating the development of ticks,resisting insecticides and improving drug resistance.Therefore,it is of great significance to study the salivary glands of ticks in order to explore the life history strategies of ticks and to find effective anti tick drugs.At present,the prevention and control of ticks mainly relies on chemical insecticides.Although the small-scale use of insecticides will not cause harm to human and animals in the short term,with the abuse of a large number of insecticides and the continuous deposition in the biological chain,it will eventually affect the health of people at the top of the food chain.Therefore,a variety of factors should be taken into account when formulating a reasonable pesticide use strategy,such as the type,concentration and use time of pesticides.Chlorpyrifos is a neutral organic non-volatile poison,which belongs to thiophosphate insecticide.It can destroy the normal nerve impulse conduction by inhibiting the activity of ache or Che in the nerves of arthropods,and cause the poisoning symptoms of arthropods.In addition,chlorpyrifos can also inhibit some other proteins in the body,such as glutathione S-transferase,phosphatase and heat shock protein.Chlorpyrifos is also a common drug to control ticks,but with the deposition of Chlorpyrifos in the food chain,it will also affect the normal development of a variety of organisms,including human beings.With the gradual delisting of chlorpyrifos,it is urgent to find other low toxicity and effective drugs to control chlorpyrifos.In this study,we analyzed the protein changes in salivary glands of H.longicornis after chlorpyrifos treatment,in order to find more molecular targets of chlorpyrifos inhibition in the body of H.longicornis.In this study,the female ticks of H.longicornis were treated with different concentrations of chlorpyrifos for different time by filter paper membrane method,and the specimens of two dimensions of different treatment concentrations and different treatment time were obtained.Furthermore,we carried out quantitative proteomics study on the salivary glands of female ticks under different treatment conditions,in order to find the target proteins affected by chlorpyrifos and the proteins involved in the detoxification process of ticks,and lay a foundation for further understanding the molecular mechanism of some regulatory processes of salivary glands of ticks under the action of chlorpyrifos.In this study,the dynamic changes of all proteins in salivary gland of H.longicornis treated with chlorpyrifos were studied by using dia quantitative proteomics.In this study,a total of 3 repeated experiments were carried out,and 13 groups of data were obtained: control0 h,control 1h,control 5 h,control 10 h,control 20 h,5M 1h,5M 5h,5M 10 h,5M 20 h,10M1h,10 M 5h,After 10 M 10h and 10 M 20h,four groups of experimental data under different extraction conditions were obtained: the effect of different concentrations of chlorpyrifos for 1h,5 h,10 h and 20 h on the protein in salivary gland of H.longicornis.There were 593 cross proteins identified in 1 h treatment group,among which 109 were differentially expressed;there were 369 cross proteins identified in 5 h treatment group,among which 55 were differentially expressed;there were 630 cross proteins identified in 10 h treatment group,among which 129 were differentially expressed;there were no significant differences in 20 h treatment group 881 proteins were identified,among which 287 proteins were differentially expressed.Bioinformatics analysis method was used to analyze these proteins,and a variety of proteins that may participate in the detoxification process were found,such as glutathione S-transferase,glutamine synthetase and a variety of heat shock proteins.The reasons for the changes of these proteins were comprehensively analyzed,which were affected by chlorpyrifos and then changed,or the necessary process to start the detoxification mechanism of the body.In addition,we also found some proteins involved in homeostasis and self-healing,such as metalloproteinases and phospholipases.RNA interference experiment was carried out on heat shock protein 70 which changed significantly under the influence of chlorpyrifos.The results showed that heat shock protein 70 was interfered,which seriously affected the blood sucking ability of H.longicornis.It indicated that heat shock protein 70 was an important protein that could help ticks resist external chlorpyrifos stimulation and enhance their own function,which was of great significance to the survival of ticks.In a word,the research results of this paper will provide new ideas for the development of tick vaccine and related drugs,and then help to formulate effective pest control strategies,which has important theoretical and practical significance.In a word,the dynamic changes of protein in salivary glands of semi-saturated female ticks under the action of chlorpyrifos were studied systematically by using DIA quantitative proteomics technology.Through the analysis of the results of quantitative proteomics,the key detoxification enzymes were finally found.The study of their functional properties and regulatory mechanism is helpful to explore the expression and regulation of many kinds of detoxification genes and provide new target genes for pest control.It can also provide new ideas for the development of tick vaccines and related drugs,and then contribute to the formulation of effective pest control strategies,which has important theoretical and practical significance. |
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