Mass production and application of activation tagged hairy root lines for functional genomic of secondary metabolism in ginseng
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Activation tagging that uses T-DNA vectors containingmultimerized transcriptional enhancers from the cauliflowermosaic virus (CaMV) 35S gene is a powerful tool todetermine gene function in plants. This approach has beensuccessfully applied in screening various types of mutationsand cloning the corresponding genes. We generated an activationtagged hairy root pool of ginseng (Panax ginseng C.A. Meyer) in an attempt to isolate genes involved in the biosyntheticpathway of ginsenoside (triterpene saponin), whichis known as the major active ingredient of the root. Quantitativeand qualitative variation of ginsenoside in activationtagged hairy root lines were profiled using LC/MS. Metabolicprofiling data enabled selection of a specific hairy root linewhich accumulated ginsenoside at a higher level than otherlines. The relative expression level of several genes of triterpenebiosynthetic pathway in the selected hairy root linewas determined by real time RT-PCR. Overall results suggestthat the activation tagged ginseng hairy root system describedin this study would be useful in isolating genes involved in acomplex metabolic pathway from genetically intractableplant species by metabolic profiling.
Abstract
Activation tagging that uses T-DNA vectors containingmultimerized transcriptional enhancers from the cauliflowermosaic virus (CaMV) 35S gene is a powerful tool todetermine gene function in plants. This approach has beensuccessfully applied in screening various types of mutationsand cloning the corresponding genes. We generated an activationtagged hairy root pool of ginseng (Panax ginseng C.A. Meyer) in an attempt to isolate genes involved in the biosyntheticpathway of ginsenoside (triterpene saponin), whichis known as the major active ingredient of the root. Quantitativeand qualitative variation of ginsenoside in activationtagged hairy root lines were profiled using LC/MS. Metabolicprofiling data enabled selection of a specific hairy root linewhich accumulated ginsenoside at a higher level than otherlines. The relative expression level of several genes of triterpenebiosynthetic pathway in the selected hairy root linewas determined by real time RT-PCR. Overall results suggestthat the activation tagged ginseng hairy root system describedin this study would be useful in isolating genes involved in acomplex metabolic pathway from genetically intractableplant species by metabolic profiling.
참고문헌 (29)
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