| Rice stripe disease, caused by Rice stripe virus(RSV), is a destructive disease in japonica ricegrown regions of China. RSV is transmitted by small brown planthopper(Laodelphax striatellus, SBPH) in a circulative, persistent and propagative manner, and also maternally from the ovary into eggs, but not in sperm. The transmission of RSV by SBPH needs to break through the obstacle of gut, salivary glands and ovary, and duplication in vector insect body. The complicated process of transmission and replication may have impact on the developmental process of SBPH. Previous researches show that the viruliferous females of SBPH usually lay fewer eggs with a longer hatch period, low hatchability, malformation and retarded or defective development compared with healthy females. Moreover, RSV reduces survival rate of 1st and 2nd instar nymphs, and also shortens the 5th instar stage and the total nymphal stage.To explore the molecular mechanism of those phenomena and understand the interaction between RSV and SBPH better, we analyzed the differential proteomics profiling of the ova between viruliferous and healthy female SBPH using an isobaric tag for relative and absolute quantitation(iTRAQ) approach, and then verified the result of iTRAQ by reverse transcript quantitative PCR(RT-qPCR). The iTRAQ result shows that 147 proteins were differentially accumulated: 98(66.7%) proteins increased, but 49(33.3%) decreased in the ova of the viruliferous females. The biological process of the differentially expression proteins is mainly about translation, metabolism, electron transport, cell cycle, transport, transcriptional electron and others. RT-qPCR was used to verify 12 differential expressed proteins from iTRAQ, showing similar trends in the transcriptional change for the 12 genes with those at the proteomic level. Differentially expressed proteins that were associated with meiosis(serine/threonineprotein phosphatase 2B and cyclin B3) and mitosis(cyclin B3 and dynein heavy chain) in viruliferous ova may influence the completion of meiosis and cleavage, which 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 disturb the supply of ATP and nutrition and then affect embryonic development.To study whether RSV has impact on the development of nymph of SBPH, we used RT-qPCR to study the transcriptional level of some development genes in SBPH after acquiring the virus. Firstly, we got 10 development-related mRNA sequences of SBPH through blast those genes sequences of Drosophila melanogaster against the SBPH transcriptome. Secondly, using the RT-qPCR, we compared the relatively transcriptional level of the 10 genes between the viruliferous and healthy nymph of SBPH. The result showed that the transcription of 8 genes showed significant difference in the condition of RSV(2 upregulated and 6 downregualted). The differentially transcriptional of dynein heavy chain and sodium/potassium-transporting ATPase subunit alpha may lead to developmental defect of nervous system and abnormal behavior. The decreased accumulation of paxillin may influence wing development. Also, five cytoskeleton proteins were downregulated, which may lead to softer exoskeleton and affect the eclosion process. In conclusion, our study of differentially translation and transcription between the viruliferous and healthy SBPH helps us to understand the interaction of RSV and SBPH better. |
PDF Full Text Request