The efficiency of RNA interference in Bursaphelenchus xylophilus.
RNA interference (RNAi) was performed on several essential genes in the pinewood nematode Bursaphelenchus xylophilus, which causes pine wilt disease. Double-stranded RNA (dsRNA) was delivered to larvae or adult worms by soaking, electroporation, or microinjection. Soaking and electroporation of L2-L3 stage worms in solutions containing dsRNA for essential genes induced over 25% lethality after 5 days, and gene-specific phenotypes were observed. This lethality agreed with significant reductions of the targeted transcripts, as assayed by reverse-transcription coupled with real time PCR. Microinjection was the most efficient route as measured by the hatching rate of F1 embryos, which was reduced by 46%. When adult worms were soaked in dsRNA, lethality was induced in the F1 larvae, revealing the persistence of knockdown phenotypes. The penetrance of the RNAi phenotypes for essential genes was relatively low but consistent, indicating that RNAi should be useful for studying the in vivo functions of B. xylophilus gene products.
Park
Jung-Eun
JE
Department of Biochemistry, College of Science, Yonsei University, Seoul 120-749, Korea.
Lee
Kyong Yun
KY
Lee
Se-Jin
SJ
Oh
Wan-Suk
WS
Jeong
Pan-Young
PY
Woo
Taeha
T
Kim
Chang-Bae
CB
Paik
Young-Ki
YK
Koo
Hyeon-Sook
HS
D000818:Animals; D018274:Electroporation; D017238:Genes, Helminth / Q000235:genetics*; D007814:Larva / Q000235:genetics / Q000378:metabolism; D008845:Microinjections; D010641:Phenotype; D028223:Pinus / Q000469:parasitology; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics*; D018510:RNA, Helminth / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017187:Tylenchida / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D014934:Wood / Q000469:parasitology
D012330:RNA, Double-Stranded; D018510:RNA, Helminth; D012333:RNA, Messenger