Abstract
Rice stripe virus-infected females of the small brown planthopper (SBPH, Laodelphax striatellus) usually lay fewer eggs with a longer hatch period, low hatchability, malformation and retarded or defective development compared with healthy females. To explore the molecular mechanism of those phenomena, we analyzed the differential proteomics profiling of the ova between viruliferous and healthy female insects using an isobaric tag for relative and absolute quantitation (iTRAQ) approach. We obtained 147 differentially accumulated proteins: 98 (66.7%) proteins increased, but 49 (33.3%) decreased in the ova of the viruliferous females. RT-qPCR was used to verify the 12 differential expressed proteins from iTRAQ, finding that trends in the transcriptional change for the 12 genes were consistent with those at the proteomic level. Differentially expressed proteins that were associated with meiosis (serine/threonine-protein phosphatase 2B and cyclin B3) and mitosis (cyclin B3 and dynein heavy chain) in viruliferous ova may contribute to low hatchability and defective or retarded development. Alterations in the abundance of proteins involved in the respiratory chain and nutrition metabolism may affect embryonic development. Our study begins to explain macroscopical developmental phenomena and explore the mechanisms by which Rice stripe virus impacts the development of SBPH.
Highlights
The fecundity of the green rice leafhopper (Nephotettix cincticeps) is significantly lower after Rice dwarf virus infection[11]
When several embryonic developmental genes of SBPH were subjected to RT-qPCR to analyze viral influence on eggs at the transcriptional level, the expression of Ls-Dorsal, Ls-Chorion peroxidase (CPO) and other 11 embryonic developmental genes differed significantly in viruliferous eggs compared with noninfected eggs
All 16 nymphs were viruliferous, indicating that the viruliferous rate (VR) of ova laid by viruliferous females was 100%, which guaranteed the availability of the viruliferous sample for subsequent experiment and analysis (Fig. 1)
Summary
The fecundity of the green rice leafhopper (Nephotettix cincticeps) is significantly lower after Rice dwarf virus infection[11]. In an RT-qPCR analysis, CYP307A1, involved in the ecdysteroid pathway, and JHAMT, involved in the juvenile hormone pathway, were found to be upregulated and downregulated, respectively, in RSV-infected 5th instars of SBPH and thought to contribute to the expedited development of the nymphs[15] Those studies have helped us to uncover the physiological and morphological influences of viruses on the vector insects, few studies have focused on proteomic changes in the host resulting from virus infection. Documenting disorders in protein accumulation in viruliferous insects can be a powerful tool for revealing the mechanisms underlying developmental changes caused by the virus Many techniques such as 2D gel based technology and isobaric tag for relative and absolute quantitation ( iTRAQ) can be used to identify variations in proteins under different conditions[16]. Study of the ova rather than the zygote can reveal the influence of RSV on SBPH from the beginning of embryonic development and exclude the interference of sperm, which cannot be infected with RSV
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