A novel, in vivo, indoor method to preserve rice black-streaked dwarf virus in small brown planthopper using wheat seedling as a bridge host

Abstract

Rice black-streaked dwarf virus (RBSDV) naturally infects Gramineae plants through small brown planthopper (SBPH) as a vector. However, RBSDV cannot be transmitted to the SBPH offspring through transovarian transmission. Wheat plant, an important intermediate host in winter, is essential for the completion of the annual cycle of RBSDV in farm ecosystem. We developed a novel, in vivo, indoor method to preserve RBSDV in SBPH using wheat seedling as a bridge host. The temperature range of 23–27°C was initially selected to rear the insects and plants. Before initiating the scheme cycle, viruliferous SBPH was obtained by feeding the virus-free 1st to 2nd instar nymphs with RBSDV-infected rice plants. Four to six RBSDV-infected SBPH were placed per plant to inoculate wheat seedlings at two-to-four leaf stages. After 48h of inoculation, the viruliferous SBPH were removed. Five mated, newly emerged virus-free SBPH females were then transferred onto each inoculated plant and allowed to lay eggs for 48h. The newly hatched SBPH were raised on wheat seedlings until the 2nd instar nymph stage, and then transferred onto healthy rice seedlings for further development until 5th instar nymphs or adults. These newly obtained viruliferous SBPH can be used for inoculating new wheat seedlings in the succeeding maintenance cycles, or for further experiments. We discovered that the incubation period of RBSDV in wheat seedlings synchronized with the gestation period of SBPH eggs at four to six inoculated viruliferous SBPH per plant and lasted for approximately seven days. In addition, this period was optimal for enhancing the SBPH infection ratio because SBPH nymphs can only acquire the virus after they hatch. The RBSDV infection ratio of the SBPHs acquired through this method consistently exceeded 50%.