presentation

1. Invitro production of Plumbagin from
Plumbago rose L. and exploration of
overproduction strategies
By ,
Sreedevi A

2. Introduction
• Plumbago rosea L. (Synonym: Plumbago indica
L.)Family: Plumbaginaceae
Common name: Laurel
• Malayalam: Chuvanna Kotuveli
• Distribution

3. • Shrubby perennial, frequently cultivated in
gardens in India
• Stems errect, trailing or climbing, simple or
branched from base, sometimes rooting at
nodes

4. Uses
• Plant yield therapeutically important
phytometabolite, plumbagin
• Roots of this plant are main source of an
alkaloids, Plumbagin
• Roots of P.rosea had been used to treat cancer
traditionally in India and Africa

5. • Is a well known microfilaricidal molecule
• Root used as abortifacient and antifertility
medicine
• Juice of leaves and roots, used for
rheumatism and paralysis, glandular
swellings, and leprosy
• Roots also used for dyspepsia, piles, diarrhea,
improve the appetite.

7. Systematic position
(According to B&H Classification
• Division - Angiospermae
• Class - Dicotyledonae
• Subclass - Gamopetalae
• Series - Inferae
• Order - Rubiales
• Family - Rubiaceae
• Genus - Odenlandia
• Species - corymbosa

8. Habitat figure

9. Importance of work
• Principle source of Plumbagin still remained in
roots collected from natural population
• Slow growth rate,absence of seeds , lack of
fruiting stage in traditional agricultural
methods necessitate search for an alternative
and effective source to meet with enhanced
commercial demand
• Has incresed market demand in both domestic
and international level

10. Objectives
• To establish suitable culture system for
production of plumbagin from P.rosea
• To screen promising germplasm for plumbagin
content
• To explore organ culture systems in
production of Plumbagin
• To optimize media factors for metabolite
production

11. • To initiate screening of cell lines and
productivity enhancement
• To explore overproduction of plumbagin
through hairy root culture, two phase culture
systems and cell immobilization
• To study effect of plumbagin precursor
supplementation in in vitro production
systems

12. • To expore role of elicitors in production of
plumbagin from various types of cultures
• To compare level of key genes associated with
plumbagin sysnthesis in control and elicitor
treated cells

13. Methodology
• Plants collected from diverse habitats
compared for plumbagin content
• Promising accessions will be maintened to
initiate in vitro cultures
• In vitro culture of P. rosea through direct and
indirect morphogenesis
• Development of callus, suspension, growth
characters of callus and suspension,
estimation of targeted metabolite production

14. • Optimization of media factors, role of elicitors,
effect of precursor(s) addition, permealizers, cell
immobilization and its effect on metabolite
production
• Cell line selection by Bergaman’s pour plate
method or FACS
• Different clones will be raised for comparing
plumbagin productivity
• Efficient clones will be screned for in vitro
production and elevated as seed culture

15. • Agrobacterium rhiogenes mediated cell
transformation and metabolite production
• Conformation of stable transformation,
evolving hairy root lines of improved
plumbagin producing lines.
• rol specific PCR amplification of transformed
lines

16. • RT-PCR to compare level of gene encoding
enzymes associated with plumbagin synthesis
• Experiment includes isolation of high quality
RNA, conversion to single stranded DNA by
reverse transcription, amplification of
plumbagin synthesizing specific gene using
specific primers

17. Possible outcome
• Method for the in vitro production of metabolite
will be evolved
• Promising germplasm will be screened for
plumbagin content
• Various factors associated with yield of
plumbagin can be identified
• Exploration of over production of plumbagin
through hairy root culture, two phase culture
systems and cell immobilization
• Comparison of key genes associated with
plumbagin synthesis in control and ellicitor
treated cells