Establishment Of Callus And Cell Suspension Culture Of Artemisia Annua L. Of Vietnamese Origin
Loading...
Date
2011
Authors
Chin, Chee Keong
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Callus cultures from three clones of Artemisia annua L. were induced from the leaf explants of axenic plantlets. MS and LV basal media supplemented with 0.5 mg/L BA, 0.5 mg/L NAA and 0.5 g/L casein hydrolysate were used as induction medium. The three clones of A. annua, namely TC1, TC2 and Highland clones, showed difference in callus growth and appearance. MS basal medium was found to be the better callus induction medium as compared to LV basal medium. Generally callus growth was better in terms of biomass yield and friability when induced on MS basal medium than it was on LV medium. Callus induction capacity of the three clones using MS medium supplemented with picloram (0-2.0 mg/L) was different. A. annua Highland clone was selected for further study as the callus could be induced consistently well in MS medium supplemented with 0.5 mg/L BA, 0.5 mg/L NAA and 0.5 g/L casein hydrolysate, and MS supplemented with 0.5 mg/L picloram. However, when callus of A. annua Highland clone was subcultured, MS medium supplemented with 2.0 mg/L picloram was found to be suitable as proliferation medium. Subsequently cell suspension culture of A. annua Highland clone was established in liquid MS medium supplemented with 2.0 mg/L picloram. Growth pattern of A. annua indicated that maximum cell growth was achieved 18 days after culture. Cell growth could be enhanced in double strength MS medium either under illumination or darkness. Photoperiod of 16-hour light was able to stimulate higher cell growth as compared to that of other photoperiods. Elicitation with arginine (4, 6, 10 mg/L) also improved cell growth. While elicitation with casein hydrolysate (0-1.0
g/L) and yeast extract (0-2.0 g/L) did not show any distinct cell growth on different feeding days. Elicitation with 2.0 g/L yeast extract one day after cell inoculation was discontinued as cell death occurred. Effect of combined elicitation of casein hydrolysate (0-1.0 g/L) and yeast extract (0-1.0 g/L) was also studied under light illumination and darkness. Cells response to combined elicitation was distinct under light illumination and under darkness. This indicated that there was interaction between light illumination and combined elicitation effect. Artemisinin was not detected in the callus and cell cultures of A. annua but sesquiterpene and other unidentified compounds were probably present. This could possibly be intermediate compounds related to artemisinin biosynthesis.
Description
Keywords
Callus cultures from three clones of Artemisia annua L , from the leaf explants of axenic plantlets