Expression of Rice Gall Dwarf Virus Outer Coat Protein Gene (S8) in Insect Cells

To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity,its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system.The S8 gene was subcloned into the pFastBacTM1 vector,to produce the recombinant baculovirus transfer vector pFB-S8.After transformation,pFB-S8 was introduced into the competent cells (E.coli DH10Bac) containing a shuttle vector,Bacmid,generating the recombinant bacmid rbpFB-S8.After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection,Sf9 cells were collected at different times and analyzed by SDS-PAGE,Western blotting and immunofluorescence microscopy.The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells.Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.

Positive correlation of methamidophos resistance between Lipaphis erysimi and Diaeretilla rapae and effects of methamidophos ingested by host insect on the parasitoid

Temporal and spatial correlated variations on the methamidophos resistance and biomolecular rate constant of acetylcholinesterase to insecticides were found between the turnip aphid, Lipaphis erysimi and its endoparasitoid, Diaeretilla rapae, collected from field colonies and an insecticide-free field insectarium in Fujian, China. Compared to the related susceptible insectarium population, L. erysimi and D. rapae displayed 7.4-29.2- and 2.6- 9.2-fold resistance ratios, respectively. In addition, two populations ofL. erysimi with different methamidophos resistance levels, that is, a field （with 5.8-fold resistance ratio） and an insectarium population, were used to study the effects of methamidophos ingested by the host insect on D. rapae development. The percentage of D. rapae cocoon formation decreased significantly when the parasitized L. erysimi were fed on cauliflower leaves treated with methamidophos at lethal concentration dosages 10 （LC50） or LC520. At LC50 dosages the percentage of D. rapae cocoon formation and adult emergence decreased significantly. When the parasitized L. erysimi were fed on methamidophos at LC50 dosage, no D. rapae cocoons were found. When the field or insectarium L. erysimi were treated with methamidophos at LC50, the susceptibility to methamidophos in the adult D. rapae emerged from the treated host insect was similar to the control. However, the susceptibility to methamidophos in the adult D. rapae became lower than the control when the host insects were treated at LC50 dosages. The data thus suggested that the methamidophos ingested by the host insect L. erysimi could be an important factor in the endoparasitoids＇ insecticide resistance development. The natural selectivity would favor the parasitoids that had developed an insensitivity to the insecticide（s）.