an age old treasure of plants for the new world’s medicine : biotechnological perspective

7/22/2019 AN AGE OLD TREASURE OF PLANTS FOR THE NEW WORLDS MEDICINE : BIOTECHNOLOGICAL PERSPECTIVE

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SHIVAJI UNIVERSITY KOLHAPURDEPARTMENT OF BOTANYPROF. DR.) G. B. DIXIT.

AN AGE OLD TREASURE OF PLANTS FOR

THE NEW WORLDS MEDICINE :

BIOTECHNOLOGICAL PERSPECTIVE


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edicated toLate Dr V N NAIK

A devoted Plant taxonomist of our country


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History of medicinal plants in

India

MEDICINAL

PLANTS

(4500 B.C.-1600 B.C.) - 67 medicinal plantshave been mentioned.

(2500 B.C. -1500 B.C.) - mention has

been made of about 290 plants used as

remedies for curing diseases.

Plants have been used for

medicinal purpose not only by

humans since prehistoric

timesbut are also used to

treat our livestock.

Rig-VedaAtharva-

Veda


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Ayurveda is UPVEDA ofAtharvaveda whose

authority was Lord

Dhanvantri, who

established the realfunction of earliest

medical science.

During this period Greek and Romans

Took substantial material from India to

enrich their medicinal knowledge

(Pal and Jain, 1998).

Higher plants have been the traditional source for

raw materials and medicinal formulations, ever since

the dawn of civilization.


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MEDICINEArt of preserving and restoring

health especially by other meansthan surgery, drugs, potions etc.

Sources of medicines

Plant Medicines

Plants

AnimalsMicrobe

sMinerals

For many centuries in all great literature, cultures

like Hindu, Buddhist, Confucian, Islamic, Roman,

Christian the practice of medicine was associated with

certain moral behaviours, qualities and values.


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The moral physician was competent, dedicated

caped confidences and cared for the sick poor

without charge. As healers gradually formed

themselves into practitioners question ofrelationships between practitioners and of

economic rivalry also

appeared as the agenda of medical ethic.

Physicians started earning more money and

thereby social prestige than the profession had

previously enjoyed. Old ideas about medical

morality were challenged by these conditions.

At mid twentieth century, the traditionalmedical morality encountered unprecedented

problems. At the same time science had brought

much more effective treatments than had everexisted.


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In general twentieth century had many

more inventions which has changed the

face of this world.To name few, Mendelian

genetics, Hybrid breeding,

mutation breeding, molecularbiology, r-DNA technology etc.

along with tremendous

development in the field ofenergy, electronics and

instrumentation.

The big challenge was of ever increasinghuman population and various ailments/

diseases like cancer, aids and other diseases

of viral origins, epidemic in nature and to date


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Thus the increasing demand and decreasing plant

resources indicate the need of multifaceted

program

develop which includes conventional and non-

conventional ways to conserve the knowledge and

In this context, zoopharmacognosy

can also play an important role (e.g.

Rubia chimpanzee interaction,

Rubiatriol, anticancer agent. Since

these primates have coexisted and

co evolved with the plants of the

tropical forests for millions of years).

,drugs occurs with increasing frequency. Crisis like

these have created a new demand for

Phytochemical screening programmes to detectantiviral plant products.


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Plant tissue culture was originally developed as aresearch tool to study the physiology and

biochemistry of plants, without the complication

of the whole-plant structure.It has made significant contributions in:

The production of plant material

Plant breeding

Gene banks

The production of chemical compounds

What is BiotechnoloBiotechnology defines any technological

application that uses a biological system, living

organisms or derivates thereof to make or modify

products or processes for specific use

(Conservation of Biodiversity).


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PLANTTISSUE

CULTURE

SEED CULTURE

ORGANCULTURE

PROTOPLASTCULTURE

EMBRYOCULTURE

CELL CULTURE

MERISTEMCULTURE

CALLUSCULTURE

BUD CULTURE


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Micropropagation

Somaticembryogen

esis

Agrobacteri

um

mediated

genetic

transformat

ion

Extraction

and

analysis ofactive

principles

from cell

cultures

ADVANCED TECHNIQUES USED IN

PLANT TISSUE CULTURE


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Micropropagation is the practice of

rapidly multiplying stock plant

material to produce a large

number of progeny plants, using

modern plant tissue culturemethods.Micropropagation is used to

multiply novel plants, such as

those that have been genetically

modified or bred through

The first commercial process to be

developed was micropropagation,which is now a worldwide industry

producing millions of plants per

annum.


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The process of initiation and development of

embryos or embryo-like structures fromsomatic cells


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AGROBACTERIUM MEDIATED

GENETIC TRANSFORMATION

A. rhizogenes causes root hairdisease (hairy root) Other bacterial

groups also contain species capable

of interkingdom genetic exchange

(Gelvin 2005).

Agrobacterium tumefaciens - or Agrobacterium

rhizogenes-mediated transformation is to date themost commonly used method for obtaining

transgenic plants.The tumorigenic host plant species for rangeA.

tumefaciens include Large number of dicots and

some monocots and Gymnosperms.Agrobacteria are naturally occurring, ubiquitous soil

borne pathogens.A. tumefaciens causes crown gall disease (tumors).


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EXTRACTION AND ANALYSIS

OF ACTIVE PRINCIPLESRenewed interest in

the therapeutic potentialof medicinal plants

means that researchers

are concerned not only

with validatingethnopharmacological usage ofplant, but also with

identifying, isolating and

characterizing the active

components.The reason that certain plantsare effective against particular

diseases is because they contain

specific chemical compounds

(the active principles)


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BIOTECHNOLOGICAL PRODUCTION

OF SECONDARY METABOLITES

Cells transformed with some of the bacterias DNA,

causes the cells to be more sensitive to the hormones

they produce. The cells form into roots. These roots

grow very fast and produce the secondary

The secondary metabolites are known to play a

major role in the adaptation of plants to theirenvironment and also represent an important source

of pharmaceuticals.

Medicinal plants are the most

exclusive sourceof life-saving drugs for majority of

the world's population.

Scientists have developed a form of root culture

usingAgrobacterium rhizogenes, the cause of hairy

root disease.


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Identification of plants at the species level is

traditionally achieved by careful examination of thespecimens macroscopic and microscopic morphology.Plant breeding, pharmacology, pharmacognosy and

other quality control disciplines

have also contributed lot in the field.

Physiological plant anatomy

(Haberlandt,1884-1945), in-vitro

culture of plant cells (White,

1939,Gauthret, 1939),

Phytochemistry (Mothes,1900-1983),

the design of bioreactors (Tulecke

Unequivocal identification and authentication of the

plants used for production is an elementary and

critical step at the beginning of an extensive quality

assurance process.

PLANT

IDENTIFICATIO

N


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Medicinal Plants: DNA-Based

Identification

Identification of plants at the specieslevel is utmost essential while making the

medicines from plants.Identifying Species (DNA-Barcoding)

Any type of organism can be identified by examinationof DNA sequences which is unique to that species.This genomic

fingerprinting can

differentiate between

individuals, species andpopulations and has

proven useful for the

characterization of sample

homogeneity and detection

DNA FINGERPRINTING

M l l bi l i l h d d f h


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Name Acronym

Polymerase chain reaction PCR

Allele-specific diagnostic PCR

Amplification refractory mutation system ARMS

Amplified fragmented length polymorphism AFLP

Arbitrarily primed PCR AP-PCR

Direct amplification of length polymorphism DALP

Multiplex PCR

PCR-selective restriction PCR-SR

Randomly amplified polymorphic DNA RAPD

Sequence characterized amplified region SCAR

Restriction length polymorphism RFLP

DNA microarray

DNA sequencing

Inter simple sequence repeat-anchored PCR ISSR-PCR

Multiplex amplification refractory mutation system MARMS

Simple sequence repeat polymorphism SSR

Molecular biological methods used for the

authentication of medicinal plants


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Nuclear and chloroplast genes used for

authentication of medicinal plantsGene Genome

18S rRNA Nuclear

Internal transcribed spacers (ITS) of 18S,

5.8S and 26S rRNA

Nuclear

Intergenic spacer of the 5S rRNA (5S gene

spacer)

Nuclear

26S rRNA NuclearatpA, atpB, atpF, atpH Chloroplast

chlB Chloroplast

matK Chloroplast

psbA, psbK, psbI Chloroplast

rbcL Chloroplast

rp14, rpl16 Chloroplast

rpoB, rpoC1 Chloroplast

rps16 Chloroplast

trnC, trnD, trnF, trnK, trnL Chloroplast

urren eve opmen n ssue cu ure ec no ogy


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urren eve opmen n ssue cu ure ec no ogyindicate that transcription factors are efficient new

molecular tools, for plant metabolic engineering to

increase the production of valuable compounds. Such

as taxol, morphine, codeine, L Dopa, Berberin,

Diosgenin, Vinblastin and Vincristine etc.Taxol : (plaxitaxol), a complex diterpene

alkaloid found in the bark of

Taxus brevifolia, is one themost promising anticancer

agents known to its unique

mode of action on the micro-

tubular cell system (Jordanand Wilson, 1995

In 1989, Christen et al., reported for

the first time the production of taxol byTaxus cell cultures various factors

influencing stability and recovery of

paclitaxol from suspension cultures by

Nguyea et al., 2001, Wu et al., 2001

Taxol

RESEARCH ACTIVITIES OF


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RESEARCH ACTIVITIES OF

OUR LABORATORYProtocols formicropropagation of

different valued plants

have been established.

Biochemical assay ofsome medicinally

important plants are

also being carried out to

assure the quality of theregenerants.

Mi ti f H id i di d R bi dif li h b


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Micropropagation of Hemidesmus indicus and Rubia cordifolia has been

established by using different phytohormones.Hemidesmus indicus Rubia cordifolia


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Progressive Stages of tissue culture in

Hem idesmus ind icus

Hardened

Rooting

Callus

Multiple shoot induction

in Hemidesmus indicus.


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Somatic

embryogenesis

in Hemidesmus

ind icus


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Rub ia cord i fo l ia

Progressive stages of tissue culture in Rubia co rdi fo l ia

Ghatge S. R., Kudale S and Dixit G. B. An Improved Plant Regeneration System

for High Frequency Multiplication of Rubia cordifolia L.: A Rare Medicinal Plant.Asian Journal of Biotechnolo 3 4 397-405 2011.


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Tissue culture in

Drimia spp.

HPLC profiles of standard Pro. A

Proscillaridin A


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PlantletRegeneration in

Chlorophy tum sp.

MS + 2,4-D

(2mg/l) + BAP

(1mg/l)

MS + 2,4-D

(2mg/l) + BAP

(1mg/l)

MS + BAP (1mg/l)

MS + 2,4-D(2mg/l) + BAP

(1mg/l)

MS +

IBA (1.5mg/l)

Saw dust +

MS basal (liqui

Soil

Protocol for in vitro multiplication of Chlorophytum sp., a valued

medicinal plant has been established.


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Tissue culture in

Grape

Fruits of Grape cv

Kedage V. V., Jai C. Tilak, G. B. Dixit, TPA Devasagayam and MinaI Mhatre Antioxidant Properties

of Different Varieties of Grapes (Vitis Vinifera L.) in India. Critical Reviews in Food Science and

-

S it bl t l f i it ti f N th d t


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Camptothecin

Suitable protocol for in vitro regeneration of Nothapodytes

nimmoniana, which is a valued medicinal plant, has been

established.

Nothapodytes nimmoniana

(Grah.) Mabb.Pai S. R., Nimbalkar M. S., Pawar N. V., Patil R. P., and Dixit G. B.Seasonal

discrepancy in phenolic content and antioxidant properties from bark of Nothapodytes

Protocol for in vitro regeneration of Ancistrocladus heynenanus


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120 DAYS MS + BAP 3.0 120 DAYS WPM + BAP

Ancistrocladus heynenanus

Wall .ex Grah.

Protocol for in vitro regeneration ofAncistrocladus heynenanus

has been established

Zingiber officinale Rosc


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Germplasm

collection

Tissue cultural stu

Zingiber officinale Rosc.

CH3O

OH

O O

H[6]Gingerol

An effective protocol for Ginger in vitro

regeneration was established and

Hardening of the tissue culture raisedplantlets was done successfully.Pawar N. V., Pai S. R., Nimbalkar M. S and Dixit G. B.RP-HPLC analysis of Phenolic Antioxidant compound 6-

gingerol from different ginger cultivars. Food Chemistry.

126: 1330-1336, 2011.

C ti f C i i th h


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Flower

Multiple

shoots

In vitrotuberformation

Conservation of Ceropegiaspecies through

Biotechnological tools

Micropropagation ofCeropegia Micropropagation ofCeropegia

panchganiensis Blatt & McCann

Agrobacterium mediated genetic transformation induced


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Agrobacterium mediated genetic transformation induced

hairy roots in Rubiacordi fo l ia and quantification of Alizarin

ALIZARIN


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DNA barcoding for identification of the

enigmatic plant Ramkand

Agave sisalana

Nimbalkar et al. (2011) DNA barcoding for identification of the enigmatic plant


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an age old treasure of plants for the new world’s medicine : biotechnological perspective

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