title: Growth and asiaticoside production in multiple shoot cultures of a medicinal herb, Centella asiatica (L.) Urban, under the influence of nutrient manipulations.
abstract: Growth and in vitro asiaticoside accumulation in multiple shoot cultures of Centella asiatica (L.) Urban was studied as a function of nutrient manipulations in the culture media. Shoot cultures raised in liquid Murashige and Skoog medium supplemented with 2.5 mg/l kinetin attained a growth index (GI) of 6.06 along with the highest asiaticoside content of 3.8 mg/g dry weight on the 35th day of the culture cycle. The shoot growth and asiaticoside accumulation were found to be influenced by the relative proportions of NH(4)(+)-N:NO(3)(-)-N or Cu(2+) concentration in the medium. Asiaticoside content in shoots increased from 5.3 to 8.9 and 8.7 mg/g dry weight when total nitrogen concentration of 60 mM in the control medium was reduced to 50 and 40 mM with a corresponding change in NH(4) (+):NO(3)(-) ratio from 20:40 to 20:30 or 20:20, respectively. Total nitrogen level higher than 60 mM drastically reduced the asiaticoside concentration in these in vitro shoot cultures. Medium devoid of Cu(2+) significantly favored higher asiaticoside accumulation in the cultured tissue (7.05 mg/g dry weight) along with an improved biomass production (GI = 7.7) when compared with shoots reared on the control medium with 0.10 μM Cu(2+) (GI = 5.8; asiaticoside content = 4.4 mg/g dry weight). Carbohydrate enrichment of the medium by increasing the sucrose concentration from 3.0 to 5.0 or 7.0% was also beneficial for biomass and asiaticoside production with GI = 17.1 and 16.9 and asiaticoside content = 7.2 and 5.2 mg/g dry weight, respectively, in comparison to control cultures maintained on medium containing 3.0% sucrose. The procedure described here provides a viable production platform for generating clean and quality material from Centella with high bioactive content.
authors.affiliation: Division of Plant Biotechnology, Central Institute of Medicinal and Aromatic Plants, Council of Scientific and Industrial Research, PO CIMAP, Lucknow 226015, India.
authors.forename: Archana
authors.initials: A
authors.lastname: Prasad
authors.forename: Archana
authors.initials: A
authors.lastname: Mathur
authors.forename: Manju
authors.initials: M
authors.lastname: Singh
authors.forename: Madan M
authors.initials: MM
authors.lastname: Gupta
authors.forename: Girish C
authors.initials: GC
authors.lastname: Uniyal
authors.forename: Raj K
authors.initials: RK
authors.lastname: Lal
authors.forename: Ajay K
authors.initials: AK
authors.lastname: Mathur
mesh_terms: D028041:Centella / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism*; D003470:Culture Media / Q000494:pharmacology; D010937:Plant Growth Regulators / Q000494:pharmacology; D014315:Triterpenes / Q000737:chemistry / Q000378:metabolism*
chemical_list: D003470:Culture Media; D010937:Plant Growth Regulators; D014315:Triterpenes; C004446:asiaticoside