Transient silencing mediated by in vitro synthesized double-stranded RNA indicates that PsCdc14 is required for sporangial development in a soybean root rot pathogen.
In many eukaryotic organisms, Cdc14 phosphatase regulates multiple biological events during anaphase and is essential for mitosis. It has been shown that Cdc14 is required for sporulation in the potato blight pathogen Phytophthora infestans; however, the role that the Cdc14 homolog (PsCdc14) plays in the soil-borne soybean root rot pathogen P. sojae remains ambiguous. PsCdc14 is highly expressed in sporulation, zoospore, and cyst life stages, but not in vegetative mycelia and infection stages, suggesting that it contributes to asexual reproduction and thus the spread of the disease. Double-stranded RNA (dsRNA) mediates gene silencing, a post-transcriptional and highly conserved process in eukaryotes, involving specific gene silencing through degradation of target mRNA. We combined in vitro dsRNA synthesis and a polyethylene glycol-mediated transformation system to construct a dsRNA-mediated transient gene silencing system; and then performed a functional analysis of PsCdc14 in P. sojae. PsCdc14 mRNA was dramatically reduced in transformants after protoplasts were exposed in in vitro synthesized PsCdc14 dsRNA, resulting in low sporangial production and abnormal development in P. sojae silencing lines. Furthermore, dsRNA-mediated transient gene silencing could enable elucidation of P. sojae rapid gene function, facilitating our understanding of the development and pathogenicity mechanisms of this oomycete fungus.
College of Plant Protection, Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education of China, Nanjing Agricultural University, Nanjing 210095, China.
Wei
W
Zhao
Xinyu
X
Yang
Suomeng
S
Dong
Yuting
Y
Sheng
Yuanchao
Y
Wang
Xiaobo
X
Zheng
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D020868:Gene Silencing; D006801:Humans; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry / Q000378:metabolism*; D010838:Phytophthora / Q000502:physiology*; D018517:Plant Roots / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D012330:RNA, Double-Stranded / Q000502:physiology*; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000382:microbiology*
D017931:DNA Primers; D012330:RNA, Double-Stranded; D010744:Phosphoric Monoester Hydrolases