Detection Of Rice Stripe Virus In Laodelphax Striatellus Fallén And Study On The Protein Related With Transmission

Rice stripe virus (RSV), a typical member of Tenuivirus, is one of important rice viruses in Easten Asian. It had inflicated damage over a huge hectarage from 2000 in Jiangsu province and its neighor areas. Polyclonal antibody SV21 and monoclonal antibody 3B9 were used in the paper. RSV in a single small brown plant hopper (Laodelphax striatellus Fallén, SBPH) could be detected with the ELISA, DIBA and Western blotting methods successfully. Results showed that the monoclonal antibody was more sensitive than polyclonal antibody and polywell-plate indirect-ELISA with polyclonal antibody or monoclonal antibody as testing antibody had a higher specificity and sensitivity than DIBA and Western blotting. Among the three molecular detection methods, RSV could be detected with 1:400 dilution of a single SBPH using RT-PCR, however, RSV could not be detected using IC-RT-PCR if the dilution of polyclonal antibody SV21 was less than 1:500 or the dilution of monoclonal antibody was less than 1:800. The results of DB-RT-PCR also showed that RSV could not be detected with less than 1:400 dilution of a single male or female SBPH.Detecting the five proteins encoded by RSV (CP, SP, NS2, NS3 and NSvc4) in SBPH, investigating the interactions between virion (CP) and the other four proteins of RSV in vitro, and providing useful data for further studying relationship between RSV and its insect vector. The total protein of virus infected SBPH was extracted, and hybrided with specific antibodies of the five RSV proteins. The gel overlay assay was performed. All the five proteins encoded by RSV presented in the SBPH. Further more, CP has a higher density among the five proteins. Besides, results indicated that there are interactions with virion (CP) and SP or virion (CP) and NSvc4 in vitro. Our study confirmed that RSV replicates in SBPH, and different concentration of the five proteins presented in SBPH body indicated the different roles in its replication. The interaction in vitro between CP and NSvc4 (the potential movement protein) indicates that the cell-to-cell movement of RSV may depend on this relationship between CP and NSvc4. Our results also showed the interaction between CP and SP, which has been reported previously on chloroplast of rice and influence to the severity of symptoms.In order to parse the case of receptor protein of RSV in SBPH . In this dissertation, through optimizing the protein of SBPH extraction methods and electrophoresis conditions, a large-scale, high- resolution and reproducible two-dimensional electrophoresis(2-DE) analysis system was established based on the technical platform of Bio-Rad large-scale 2- DE system. At least 54 protein pots were detected on the gels of SBPH.The protein of Mr= 60 KD, pI=6.7 was detected by virus overlay assay (VOA) from SBPH of non-infected and high-affinity. There is interaction with the protein and RSV in vitro, specially.