Transcript Plants as source of biologically active substances


 Plants are producers of many BAS –
substances, capable to affect on
biological processes in an organism.
For ex., glycosides, saponins, sterols,
carothenoids, polyphenols, alkaloids,
vitamins, quinones, and also the
substances possessing specific aroma,
taste and coloring. Biologically active
agents are products of a secondary
methabolism which are called
biosynthesis by-products. Now it is
known more than 100 000 secondary
metabolites produced by plants.
Many of them are practically,
economically important products and
are used in the pharmacological,
cosmetic, food industry.
Industrial use of some plant products as
pharmaceutical remedies

Plant product
Plant species
Codein (alcaloid)
Papaver somniferum
Diosgenin (steroid)
Dioscorea deltoidea
Quinin (alcaloid)
Cinchona ledgeriana
Digoxin (cardiosteroid)
Digitalias lanata
Scopolamin (alcaloid)
Datura stramonium
Vincristin (alcaloid)
Catharanthus roseus
Ten most used medicinal substances,
received from plants
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API
Steroids from diosgenin
Action
Contraceptive
Source plant
Dioscorea deltoidea
Codein
Pain-killer
Papaver somniferum
Atropin
Anticholinergic
Atropa belladonna L.
Reserpin
Hipotensive
Rauwolfia serpentina L.
Hyosciamin
Anticholinergic
Hyoscyamus niger L.
Digoxin
Cardiac stimulant
Digitalis lanata L.
Scopolamin
Anticholinergic
Datura metel L.
Digitoxin
Cardio-vascular
Digitalis purpurea L.
Pilocarpin
Cholinergic
Pilocarpus jabonandi
Chinidin
Antimalarial
Cinchona ledgeriana
Examples of medicinal substances received on a
basis of callus cultures
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Steviozid is natural sweetener and
sugar substitute, is successfully used
instead of artificial sweetening
substances. Initial plant is Stevia
rebaudiana Bertoni.
Arglabin is antineoplastic
substance. Initial plant is Artemisia
glabella Kar. et Kir. Is a part of the
preparation of the same name.
Lappakonitin is diterpen alkaloid,
antiaritmic. Initial plant is Aconitum
septentrionale Koelle. Is a part of
Allapinin's preparation
Transgenic agricultures

corn
soya
 with not set effects of manifestation of a
gene (possibility of change of a chemical
composition and decrease in a nutrition
value of a product).
 change of the technological parameters
worsening consumer properties of
products.
 as a result of gene modification any
components causing allergic reactions
can be synthesized, or to appear the
dangerous connections possessing
mutagen, cancerogenic or toxic effect.
cotton
rape
potato
tomato
Biotransformation of cardenolids
 Plants of a foxglove (Digitalis lanata) in a large number
synthesize digitoxin instead of necessary digoxin. For the
corresponding biotransformation with success use
undifferentiated suspension culture of a foxglove. The
immobilized cells of this culture are capable to transform
long time with a constant speed β-methyligitoxin to βmethyldigoxin.
 "+" uses of suspension cultures for synthesis of secondary
metabolites commercially
• economic benefit in receiving cheaper production in the
planned quantities.
• exception of destruction of rare wild-growing plants.
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Perspective directions:
selection of specialized
populations of cells
optimization of conditions of
cultivation.
improvement of methods of
biotransformation of
metabolites and an
immobilization of cultivated
cells
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Panax
Berberis vulgaris L
Rauwolfia serpentina Benth
source of hypotensive and
antiarhythmic
indol alkaloids. Methods of the
cellular
selections with use of the
chemical
mutagens the highly
productive strain accumulating
antiarhythmic
alkaloid aimalin was received.
Cell strain of Panax ginseng Thalictrum minus L. 50% from the sum of alkaloids,
synthesized by culture.
Directions of creation of new technologies on
the basis of cultivated fabrics and cells of plants
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 Receiving biologically active agents of a phytogenesis: traditional products of a secondary metabolism (toxins,
herbicides, regulators of growth, alkaloids, steroids,
therpenoids, having medical application).
 Cells cultivated in suspension can will be applied as the
multifermental systems capable to a wide range of
biotransformations of chemicals (reaction of oxidation,
restoration, a hydroxylation, methylation, demethylation,
glikolisation, isomerization).
 The accelerated clonal microreproduction of plants
allowing from one explant to receive from 10000 to
1000000 plants a year, and all of them will be genetically
identical.
Types of cell and tissue cultures
Callus (superficial cultivation):
 Dense fabrics with zones of reduction of a cambium and vessels
 Fabrics of average density with well expressed meristematic centers
(organogenesis)
 The friable fabrics which have been strongly flooded, easily breaking up to
separate cells (receiving suspension)
Suspension (deep cultivation)
 suspensions from callus tissues of friable type (or dense having excluded
from a nutrient medium of salt of Sa ² +), it is possible to cultivate tissue
on the medium containing auxin, 2,4-D or enzymes – pectinase (0,2 mg/l)
and cellulose (0,01 mg/l).
 suspensions from an explant on a method F. Stewart - an explant place on
liquid Wednesday at continuous automatic hashing, dedifferentiation cells
come off an explant, forming suspension in a nutrient medium.
Continuous stirring – a necessary condition of cultivation of cellular
suspensions. suspension cultures are presented by different units of callus
cells.
Applied aspects of suspension cultures:
 receiving the isolated protoplasts for cellular selection and cellular
engineering (introduction of alien DNA)
 for receiving secondary metabolites, identification of new substances, for
cultivation of a cellular biomass.
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Asynchrony reasons:
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 1 . Features of a look, grade, genotype of an individual plant, and
feature of an explant.
 2 . Cultivation stresses, for example non-optimal for this type of
cells Wednesday.
 3 . Change of balance of endogenous hormones and concentration
in the environment of exogenous hormones during cultivation.
 4 . Genetic heterogeneity of cells and clones.
 5 . Anomaly of a mitotic cycle of cells of in vitro.
 6 . Physical factors (temperature, light, aeration).
 Asynchrony – steady property of population of callus cells.
 If by means of specific influences to synchronize proliferation of
cells of population, already through 3 - 4 divisions it again to
become asynchronous.
Characteristic of callus cells
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 The callus cell has the cycle of development similar to a cycle of
other cells.
 features physiological plants peculiar to cells - frost resistance,
resistance to abiotic factors (temperature, salinization,
photoperiodic reaction), ability to synthesis of secondary
metabolites.
 features characteristic for callus - it is long cultivated in vitro of a
cell of the highest plants (callus and suspension), form specific
population of the somatic cells, being characterized physiological
asynchrony and genetic heterogeneity.
Physiological asynchrony (the most important property of non-sexual
population) - in each this timepoint of a cell are in different stages of
growth: one share, others grow, and the third already grow old.
General physiological condition of population estimate on a condition
of the majority of cells.
Hormone-resistance
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 autonomy after the relation to auxins and citoquinins, arising at long
cultivation practically at all fabrics.
 Fabrics can grow on the Wednesday without hormones, as tumoral cells
unlike normal callus fabrics. Externally gormonnezavisimy fabrics differ
nothing from the callus.
 Though hormones also cause mutations, callus fabrics from the majority of
plants are formed only at presence at a nutrient medium and auxins, and
citoquinins.
 cells which in the course of cultivation gained property of a hormoneresistance, are called "got used".
 The fabrics formed by "got used" cells, call "chemical tumors" (unlike
vegetable or genetic tumors).
 Genetic tumors arise on interspecific hybrids of plants.
 Vegetable tumors have a bacterial or virus origin. Most often vegetable
tumors arise at hit in plants of agrobacteria.
Microclonal propagation
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 Similarly, the type of vegetative propagation of plants, but takes
place in vitro, with cells isolated from tissues can get a lot of new
plants. Prerequisite - the identity of the original plant material
obtained the mother plant. More recently, it is seen as a possible way
to accelerate cloning vegetatively propagated plants, as well as an
auxiliary method of virus free plants. However, some studies have
shown that the value of this method considerably increases for clonal
plant breeding (experimental mutagenesis and dechimaeration),
cryopreservation of valuable source material, as well as several
others. The ability to produce large amounts (a few million or more)
of somatic embryos in vitro conditions used to develop the
technology of mass and continuous production of "artificial seeds".
Furthermore, the method of clonal micropropagation can be
successfully used to create synthetic varieties. To date, the number of
species that can be cloned in vitro is about 1000.
Avantages
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 Considerably higher rates of reproduction (take 100 000-1 000
000 meriklones a year, whereas in the normal reproduction - 5100 plants during the same period);
 Miniaturization process, leading to economies of areas
occupied by uterine and propagated plants;
 Improvement of plants from fungal and bacterial pathogens,
viruses, mycoplasma, viroid and nematode infections;
 Possibility of multiplication and rooting of plant breeders is
difficult by conventional methods.
Although this method of micropropagation of plants is quite time
consuming and expensive, in some cases based on it has become
possible to create economically viable biotechnology.
Hairy roots
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Native roots of
Scutellaria baicalensis
Georgi.
Hairy roots of
Scutellaria baicalensis
Georgi.
Morphology of hairy roots
Rubia tinctorum grown in
Murasige-Scug and B-5
Rauwolfia in flask
Scutellaria baicalensis
hairy root
Yong hairy roots of Rauwolfia
freeze-dried hairy
roots of Rubia
tinctorum
Salvia officinalis in flask
Hairy roots of Salvia officinalis L.: branching
Anatomy of hairy roots
Powder of hairy roots of Rubia tinctorium
in floroglucin 1,5*20*2
Hairy roots of Salvia officinalis L
1,5*20*2
Sucking fuzzes
root cap
axon
suberic
Lignificated
tissuies
Subericпробки
cells
Клетки
tracheids
Sucking fuzzes
Suberic cells
Hairy root of Sophora sp. in floroglucin 1,5*20*2
transverse section of hairy
roots of Hedysarum theinum in
floroglucin 1,5*40*2