Camptothecin (CPT) & 10-hydroxycamptothecin (HCPT)
Prospects of Campthoeca Organ culture
Objective of the study
Four seedlings were randomly chosen from each sources for further cultivation (Ch 1 = Bp31, Bp81, Bp101, Bp141), (Ch 2 = Mp27,Mp28,Mp35,Mp36), (Lou = Lp4, Lp13b, Lp18, Lp45).
Embryogenic calli and somatic embryos were cultured on half strength MS medium
supplemented with 2 mg/l BAP plus 0.1 mg /l IAA.
Shoot tips were excised and subcultured for a period of 8 weeks in TIS and on solid medium.
Explants were cultured in both the Dual-Vessel System (DVS) and RITA vessel as previously described containing a full strength MS medium fortified with 0.5 mg/l BAP plus 30 g/l sucrose
During the first 4 weeks 200 ml culture medium was used in DVS and thereafter 400 ml and for the RITA vessel a volume of 250 ml was used throughout this study
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Production of Secondary metabolites through Organ culture
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All cultures were of the same age and were maintained under a 16-h photoperiod at 25 ± 1C.
Collected into a 50 ml centrifuge glass tube to which 5 ml of 61% ethanol was added.
The extract was centrifuged at 15,000g for 10 min
and the supernatant was filtered through a filter into HPLC vials for CPT analysis.
Determination of CPT by HPLC
All analyses were performed on an isocratic reverse-phase high performance liquid chromatography system (RP-HPLC).
Objective of the study
To evaluate the effect of different additives on SLS induction and crocin production Sodium acetate (SA), serine and glycine were added
activated charcoal and PVP (anti browning agents) , All media contained 6% (w/v) sugar and were solidified with agar (0.6%, w/v)
Culture was maintained at room temperature in the dark, Effect of light on induction was also studied
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Production of Secondary metabolites
SLS formed on the explants were excised and dried at 40⁰C for 8 h
Crocin extraction : methanol extraction
Crocin in all the extraction samples were quantified by the reverse-phase HPLC
Shikonin Derivatives
Objective of the study
Transgenic root lines and culture conditions
Determination of growth and shikonin derivatives Accumulation
Treatment for enhancement of shikonin derivatives production
Conclusions
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