Studies On Rice Grassy Stunt Virus Effects On Wing Dimorphism Of The Brown Planthopper Nilaparvata Lugens

Grassy stunt disease of rice caused by Rice grassy stunt virus (RGSV) is one of the severe diseases of rice in tropical and subtropical rice-growing areas. RGSV is a member of the genus Tenuivirus, and is transmitted by brown planthopper (BPH, Nilaparvata lugens) in persistent and proliferous manner. The brown planthopper is a long-distance migration insect with wing dimorphism. Previous studies have shown that virus can affect wing dimorphism of BPH. In this study, we focused on how RGSV influence wing dimorphism of BPH by different experiments, including the biological statistical experiment, real-time PCR, yeast two hybrid experiment, subcellular co-localization and prokaryotic expression experiment. The results are summarized as below:1. In the biological statistical experiment, the wing dimorphism of RGSV-infected BPH had significantly different compared with healthy BPH. The ratio of brachypterous and macropterous BPH adults was 23.17±0.56 and 60.43±0.22 percent in healthy BPH adults, whereas the ratio was 36.26±0.42 and 52.32±1.16 percent in RGSV-infected BPH adults.2. Expression analysis of the relevant genes involving in wing dimorphism of BPH adults by using real-time PCR. The results showed that all tested genes, including flightin, Tnc, Titin and LN-a2, was down-regualted.3. The construction of yeast expression vector, pGADT7-flightin, and the performance of yeast two hybrid experiment. The results indicated that flightin of BPH can only interact with P1, P3 and P5 proteins of RGSV.4. The construction of transient expression vector, pEarly gate101-flightin and pEarly gate102-flightin, and the performance of subcellular co-localization experiment. The results showed that flightin of BPH can interact with P1, P3 and P5 proteins of RGSV, were consistent with the yeast two hybrid results.5. The construction of prokaryotic expression vector, pDEST17-flightin, and the preparation of polyclonal antibody of flightin. Western blotting analysis indicated that the flightin antibodies can specifically react with flightin protein in BPH. The ELISA assay showed that the potency of antiserum was 1:6400.In conclusion, these results illustrate the relationship between RGSV and wing dimorphism of BPH. And some key genes were identified through some related molecular biology experiments, which lay a good foundation for further research.