概要: Ethylene plays critical roles in plant development and biotic stress response. However, the mechanism of ethylene in host antiviral response remains unclear. Rice dwarf virus (RDV) triggers the ethylene production resulting in host susceptibility by enhancing the activity of S-adenosyl-L-methionine synthase (SAMS) in ethylene synthesis pathway. Our molecular and genetic data demonstrated that Pns11 specifically interacted with OsSAMS1 to enhance its enzymatic activity, resulting in increased production of ethylene in both Pns11 overexpressing and RDV-infected rice. Treatment of rice plants with ethylene precursor increased their susceptibility to RDV infection. More importantly, knockout of OsSAMS1 (CRISPR/Cas9) or blocking ethylene signaling in an ethylene-insensitive mutant (osein2) decreased susceptibility to RDV infection, and overexpression of OsSAMS1 or Pns11 increased susceptibility. In this study, we have analyzed the relationship between ethylene pathway response and RDV infection or OsSAMS1 and S11 transgene by the gene expression analysis through the RNA-seq.
项目整体设计: Comparison between Mock and RDV infected rice, 3 control, 3 RDV-infected, Comparison between WT and OsSAMS1 and S11 transgenic rice , 2 control, 3 OsSAMS1 OX, 3 Ossams1 KO, 3 S11 OX, independently grown and harvested, 1 sample derived from 3 plants grown under same conditions.
Rcie seedlings were grown in a greenhouse at 28-30℃ with natural light conditions.
处理方案:
Mock: 14 days old seedlings were inoculated with 3 virus free insect per plant. RDV: 14 days old seedlings were inoculated with 3 viruliferous insect per plant. WT and OsSAMS1 and S11 transgenic rice: grown in a greenhouse at 28-30℃ with natural light conditions without any inoculation.
提取方案:
Total RNA were extracted from 42 days old seedlings using Qiagen RNeasy plant mini kit, one sample pooled from 3 individual palnts.
建库方案:
RNA libraries were prepared for sequencing using standard dUTP strand-specific RNA-seq library protocols
