Abstract
Polyphagous Apolygus lucorum has become the dominant insect in Bacillus thuringiensis (Bt) cotton fields. Hormone 20-hydroxyecdysone (20E) regulates multiple insect development and physiology events. 20E responses are controlled by pathways triggered by phospholipase C (PLC)-associated proteins. However, 20E-modulated genes and related proteins that can be affected by PLC still remain unknown. Here, isobaric tag for relative and absolute quantitation (iTRAQ) and immunoblotting techniques were used to compare differentially expressed proteins (DEPs) in A. lucorum in response to the treatment of 20E and the PLC inhibitor U73122 as well as their combination. A total of 1,624 non-redundant proteins and 97, 248, 266 DEPs were identified in the 20E/control, U73122/control, and 20E + U73122/control groups, respectively. Only 8 DEPs, including pathogenesis-related protein 5-like, cuticle protein 19.8, trans-sialidase, larval cuticle protein A2B-like, cathepsin L1, hemolymph juvenile hormone-binding protein, ATP-dependent RNA helicase p62-like, and myosin-9 isoform X1, were detected in all three groups. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the DEPs were involved in diverse signaling pathways. The results were validated by immunoblotting, which highlighted the reliability of proteomics analysis. These findings provided novel insights into the function of PLC in 20E signaling pathway in A. lucorum.
Highlights
Apolygus lucorum (Meyer-Dür) is an agricultural pest found in northern China, damaging more than 200 plant species (Lu and Wu, 2011; Pan et al, 2013)
The isobaric tag for relative and absolute quantitation (iTRAQ) approach was employed to perform comparative analysis among the three groups to obtain a global view of proteome alteration in A. lucorum nymphs in response to 20E, U73122, and 20E + U73122
The combination of two-dimensional polyacrylamide gel electrophoresis (2-DE), MS, peptide sequencing, and database search has been a great tool for determining differentially expressed proteins (DEPs), but is relatively complex and unsuitable for quantitating proteins in minute quantities. iTRAQ, a novel stable isotope technique for protein measurement by employing mass spectrometry (Zieske, 2006; Pierce et al, 2008), could comparatively assess eight distinct specimens concurrently and examine proteome profile changes caused by various treatments
Summary
Apolygus lucorum (Meyer-Dür) is an agricultural pest found in northern China, damaging more than 200 plant species (Lu and Wu, 2011; Pan et al, 2013). Meyer-Dür adults have high dispersal ability and severely threaten the cotton, and many fruit and vegetable industries. Proteomic Profiling in Apolygus lucorum crops (Lu et al, 2010). Since 2010, the cotton-planted areas have decreased sharply, with the expansion of maize and some other crops, such as vegetables, soybeans, and peanuts. A significant increase in the availability of host plants might have a pronounced effect on A. lucorum densities in the local region (Lu and Wu, 2008). Understanding the regulatory mechanisms underlying insect growth and development is crucial for scientific prevention, control, and integrated management of pests. There is an urgent need to develop new pest management strategies
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