Identification of regulatory protein genes involved in alkaloid biosynthesis using a transient RNAi system.
RNAi (RNA interference, RNA silencing) is a powerful tool in functional genomics. We report here the use of transient RNAi to isolate regulatory factor genes involved in isoquinoline alkaloid biosynthesis in Coptis japonica protoplasts. Double-stranded (ds) RNAs prepared against candidate regulatory factors, which were predicted from an EST library, were introduced into C. japonica protoplasts by polyethylene glycol (PEG)-mediated transformation, and their effects were monitored by real-time reverse transcription (RT)-polymerase chain reaction (PCR). The potential of this transient RNAi system to characterize the functions of regulatory factor genes in alkaloid research is discussed.
Yamada
Yasuyuki
Y
Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
Kato
Nobuhiko
N
Kokabu
Yasuhisa
Y
Luo
Qingyun
Q
Dubouzet
Joseph Gogo
JG
Sato
Fumihiko
F
D000470:Alkaloids / Q000096:biosynthesis*; D039323:Coptis / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D055785:Gene Knockdown Techniques / Q000379:methods*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D011523:Protoplasts / Q000378:metabolism; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D000470:Alkaloids; D010940:Plant Proteins; D012330:RNA, Double-Stranded; D012333:RNA, Messenger; D014157:Transcription Factors