Enhanced production of asiaticoside from hairy root cultures of Centella asiatica (L.) Urban elicited by methyl jasmonate.
Transformed root ("hairy root") cultures have been shown to be a good model for the study of many secondary metabolites. However, economically important compounds such as asiaticoside and madecassoside are produced in insignificant amounts in the root of Centella asiatica (L.) Urban. To overcome this problem, C. asiatica was transformed using Agrobacterium rhizogenes strain R1000 that harbors pCAMBIA1302 encoding the hygromycin phosphotransferase (hpt) and green fluorescence protein (mgfp5) genes and the hairy culture was coupled with elicitation technique. Hairy roots were obtained at a frequency of up to 14.1% from a tissue junction between the leaf and petiole. Abundant hairy roots were observed when co-cultivation of the plant with A. rhizogenes was done for 7 days (36.1%). Transformation was confirmed by PCR and Southern blot analyses. Five weeks after inoculation, no asiaticoside was detected in the hairy root samples. However, when 0.1 mM methyl jasmonate (MJ) was applied as an elicitor to the culture medium for 3 weeks, a large quantity of asiaticoside was generated (7.12 mg/g, dry wt). In the case of gene expression, 12 h after MJ treatment the expression of the CabAS (C. asiatica putative beta-amyrin synthase) gene in the hairy roots is significantly different from that of the control and this level of transcripts was maintained for 14 days. Our results showed that production of C. asiatica hairy roots could be optimized and the resulting cultures could be elicited with MJ treatment for enhanced production of asiaticoside.
Kim
Ok-Tae
OT
Division of Ginseng and Medicinal Crop, National Institute of Crop Sciences, RDA, Suwon 441-857, South Korea.
Bang
Kyong-Hwan
KH
Shin
Yu-Su
YS
Lee
Min-Jeong
MJ
Jung
Su-Jin
SJ
Hyun
Dong-Yun
DY
Kim
Young-Chang
YC
Seong
Nak-Sul
NS
Cha
Seon-Woo
SW
Hwang
Baik
B
D000085:Acetates / Q000494:pharmacology*; D002478:Cells, Cultured; D028041:Centella / Q000187:drug effects / Q000378:metabolism*; D003517:Cyclopentanes / Q000494:pharmacology*; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D019751:Intramolecular Transferases / Q000378:metabolism; D054883:Oxylipins / Q000494:pharmacology*; D010937:Plant Growth Regulators / Q000494:pharmacology; D010940:Plant Proteins / Q000378:metabolism; D018517:Plant Roots / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D012231:Rhizobium / Q000235:genetics; D014315:Triterpenes / Q000378:metabolism*
D000085:Acetates; D003517:Cyclopentanes; D054883:Oxylipins; D010937:Plant Growth Regulators; D010940:Plant Proteins; D014315:Triterpenes; C072239:methyl jasmonate; D019751:Intramolecular Transferases; C039924:2,3-oxidosqualene-beta-amyrin-cyclase; C004446:asiaticoside