Rice cultivar Oryza sativa L. japonica. Nipponbare (NIP) was used in this study. The cultivar Nipponbare is highly susceptible to SRBSDV and RSV (Xu et al., 2014 (link); Yang et al., 2018b (link)). NIP was used to produce transgenic rice. SRBSDV-infected plants were kindly provided by Professor Guohui Zhou and Tong Zhang (South China Agricultural University Guangzhou, China). Rice seedlings were grown in a greenhouse at 26 to 28°C with a 14-h light/10-h dark cycle under artificial light. Rice plants infected with SRBSDV and RSV were cultivated in an experimental field in Changsha and Nanjing, respectively, under natural long-day conditions. Viruliferous or virus-free planthoppers were reared on healthy rice seedlings (Wuyujing No. 3) in glass beakers at 25°C.
SRBSDV was transmitted by the white backed planthopper (Sogatella furcifera) at approximately the 1.5-leaf-stage of rice seedling. To obtain viruliferous insects, nymphs were reared on virus-infected rice plants for 2 days, and viruliferous or virus-free nymphs were transferred to each experimental rice plant to feed for 3 days, after which the planthoppers were removed. The proportions of healthy plants were calculated 30 days after inoculation. The percentage of about 30 plants infected by virus (viral incidence) of each of triplication was determined following specific quantitative RT−PCR of samples of each plant using virus-specific primers (Table S1
RSV was transmitted experimentally to rice plants by the small brown planthopper (Laodelphax striatellus) at approximately the 1.5-leaf-stage of rice seedling. To obtain viruliferous insects, nymphs were reared on virus-infected rice plants for 2 days, and viruliferous or virus-free nymphs were transferred to each experimental rice plant to feed for 3 days, after which the planthoppers were removed. The proportions of healthy plants were calculated 30 days after inoculation. The percentage of about 30 plants infected by virus (viral incidence) of each of triplication was determined following specific quantitative RT−PCR of samples of each plant using virus-specific primers (Table S1
SRBSDV was transmitted by the white backed planthopper (Sogatella furcifera) at approximately the 1.5-leaf-stage of rice seedling. To obtain viruliferous insects, nymphs were reared on virus-infected rice plants for 2 days, and viruliferous or virus-free nymphs were transferred to each experimental rice plant to feed for 3 days, after which the planthoppers were removed. The proportions of healthy plants were calculated 30 days after inoculation. The percentage of about 30 plants infected by virus (viral incidence) of each of triplication was determined following specific quantitative RT−PCR of samples of each plant using virus-specific primers (
RSV was transmitted experimentally to rice plants by the small brown planthopper (Laodelphax striatellus) at approximately the 1.5-leaf-stage of rice seedling. To obtain viruliferous insects, nymphs were reared on virus-infected rice plants for 2 days, and viruliferous or virus-free nymphs were transferred to each experimental rice plant to feed for 3 days, after which the planthoppers were removed. The proportions of healthy plants were calculated 30 days after inoculation. The percentage of about 30 plants infected by virus (viral incidence) of each of triplication was determined following specific quantitative RT−PCR of samples of each plant using virus-specific primers (
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