cultivation and collection of drugs from natuaral origin

1. Cultivation and collection
of drugs of natural origin

2. Cultivation of Crude Drugs:
Cultivation of medicinal plants requires intensive care and management. The conditions and duration of cultivation
required vary depending on the quality of medicinal plant materials required.
Methods of Propagation:
I. Vegetative propagation (Asexual propagation):
Vegetative propagation can be defined as regeneration or formation of a new individual from any vegetative part of
the plant body. The method of vegetative propagation involves separation of a part of plant body, which develops into a
new plant.
VEGETATIVE PLANT PART EXAMPLE :
1. Bulb Allium 2. Tuber Potato 3. Offset Valerian 4. Rhizome Ginger and Haldi 5. Vegetative propagation by root
Asparagus 6. Corms Colchicum 7. Runner Peppermint.
Other Vegetative propagation methods:
1. Cutting: These are the parts of the plant (stem, root or leaf) which, if grown under suitable’ conditions, develop
new plants. Stem cutting are generally used to obtained new plants. Examples: Sugarcane and rose, etc.
2. Layering: Roots are induced on the stem while it is still attached to the parent plant. This part of stem is later
detached from the parent plant and grown into a new plant. Examples: Jasmine plant

3. 3. Grafting: New variety is produced by joining parts of two different plants. The rooted shoot of one plant, called
stock, is joined with a piece of shoot of another plant known as scion. Examples: Rose, citrus and rubber, etc.
Advantages of Asexual Propagation:
•As resultant species formed through asexual process are genetically identical, useful traits can be preserved among
them.
•Asexual propagation allows propagation of crops that do not possess seeds or those which are not possible to grow
from seeds. For e.g. Jasmine, sugarcane, potato, banana, rose etc.Plants grown through vegetative propagation bear
fruits early.
•In this type, only a single parent is required and thus it eliminates the need for propagation mechanisms such as
pollination, cross pollination etc.
•The process is faster than sexual propagation. This helps in rapid generation of crops which in turn balances the loss.
Disadvantages of Asexual Propagation
•Diversity is lost in asexual propagation which is the main reason behind occurrence of diseases in future plant
species.
•As many crops are produced with this process, it leads to overcrowding & lack of nutrients.
•New varieties of crops cannot be developed in this type of propagation.
•Asexual propagation is an expensive process that requires special skills for successful cultivation of crops.
•Crops produced through this process have shorter life-span than those grown through sexual process.
•Species involved in this process are less likely to resist pests and diseases.

4. Importance of Asexual propagation:
1. It is a cheaper, easier and rapid method of multiplication. Many fruit trees usually require 4-5 years to bear the
fruits when developed from seeds. The plants developed by vegetative methods, take only a year to bear fruits.
2. Plants like roses and chrysanthemum, etc do not form viable seeds. Thus, vegetative propagation is the only method
of propagation is the only method of reproduction and continuation of species in such plants.
3. All the plants developed by these methods will be generally similar to the parent plant.
II. Sexual Propagation:
The process of sexual propagation:
(i) Pollination: This is the transfer of pollen grains from the anther to the stigma.
(ii) Fertilization: Fusion of male and female gametes takes place, resulting in the formation of zygote.
(iii)Seedling: Multiplication of plants by using seed is called as seed propagation

5. Dormancy: It is term used to describe a seed that will not germinate because of any condition associated either with the
seed itself or with existing environmental factors such as temperature and moisture.
Rest Period: Some seeds will not germinate immediately after harvest even if conditions are favorable. This failure to
germinate is due to physiological condition. This is said to be the seeds are in the rest period.
Seed viability and longevity: Viability means the presence of life in the seed. Longevity refers to the length of time that
seeds will retain their viability.Some seeds are short lived. (Citrus).
Pre – germination Seed Treatments to improve germination rate:
1. Chemical (Acid scarification): The purpose is to modify hard or important or impermeable seed covering
generally soaking seed in concentrated sulphuric acid is an effective method. The time of treatment may vary from
10 minutes to 6 hour according to species sometimes phyto hormones like gibberellins, ethylene, Cytokinins are also
used. Other chemicals like Potassium nitrate, Thiourea, Sodium hypochlorite also used.
2. Mechanical (Scarification): Seeds of a few species with impermeable seed coat. i.e. hard seed coat can be rendered
permeable to water and gases their germination is greatly improved by mechanical scarification in taking care that
seeds should be injured *not be injured heavily. This can be achieved by i) Placing the seeds between two sand
paper doses, one station and other revolving. ii) Passing seeds through machine that scratches the surface. iii) Filling
and notching to make the seed coat permeable to water.

6. 3. Seedling (Boiled Water Treatment): Pouring boiling water over seeds and getting it to cool gradually for about 12
hour to soften hard shelled seeds. E.g. Coffee.
4. Soaking in Water: The purpose of soaking seeds in water is to modify hard seed coats, to remove inhibitors to soften
seed and to reduce the time of germination. The time of soaking seeds in cold water depend upon the hardness of the
seed coat. E.g beans.
5. Moist Chilling: Seed of many woody trees or shrubs are exposed to low temperature to bring about prompt and
uniform germination. Germination rate measured by 1. Rolled towel test 2. Excised embryo test
Advantages of Sexual Propagation:
•Simplest, easiest and the most economical process
•Some plants, trees, vegetables or fruits species can propagate only through sexual propagation.
E.g. – marigold, papaya, tomato.
•This type of propagation leads to better crop species that are stronger, disease- resistant and have longer life-span.
•Viral transmission can be prevented in this type of propagation.
•It is the only propagation process in which resultant offspring have genetic variation and exhibit diversity of
characters from parent crops.
•This genetic variation is responsible for continuous evolution that keeps on producing better & better offspring.
•Easy storage and transportation of seeds.

7. Disadvantages of Sexual Propagation:
•Seeds take a long time to turn into mature plants i.e. time interval between sowing and flowering is longer.
•Seedlings propagated through sexual propagation are unlikely to have same genetic characteristics as that of parent
plants
•Some plant species do not produce viable seeds through sexual propagation and hence are unsuitable to propagate for
the same.
•Plants that do not have seeds can’t be propagated through this process.
•There are many factors that can affect the viability of seeds, including moisture, air, temperature, and light.
III. Micro propagation / Plant Tissue Culture:
This method consists of growing cell, tissue and organ in culture. Small pieces of plant organs or tissues are grown in a
container with suitable nutrient medium, under sterilized conditions. The tissue grows into a mass of undifferentiated
cells called callus which later differentiates into plantlets. These are then transferred into pots or nursery beds and
allowed to grow into full plants. Plant tissue culture is widely used to produce clones of a plant in a method known as
micropropagation to conserve rare or endangered plant species. Micro propagation is useful in raising disease free
plants, homozygous diploids, and those without viable seeds

8. COLLECTION OF DRUGS:
• Medicinal plant materials should be collected during the appropriate season or time period to ensure the best
possible quality of both source materials and finished products.
•It is well known that the quantitative concentration of biologically active constituents varies with the stage of plant
growth and development.
•The best time for collection (quality peak season or time of day) should be determined according to the quality and
quantity of biologically active constituents rather than the total vegetative yield of the targeted medicinal plant parts.
•In general, the collected raw medicinal plant materials should not come into direct contact with the soil. If
underground parts (such as the roots) are used, any adhering soil should be removed from the plants as soon as they
are collected.
•Collected material should be placed in clean baskets, mesh bags, other well aerated containers.
•After collection, the raw medicinal plant materials may be subjected to appropriate preliminary processing, including
elimination of undesirable materials and contaminants, washing (to remove excess soil), sorting and cutting.
•The collected medicinal plant materials should be protected from insects, rodents, birds and other pests, and from
livestock and domestic animals.
•If the collection site is located some distance from processing facilities, it may be necessary to air or sun-dry the raw
medicinal plant materials prior to transport.
•If more than one medicinal plant part is to be collected, the different plant species or plant materials should be
gathered separately and transported in separate containers. Cross-contamination should be avoided at all times.
•Collecting implements, such as machetes, shears, saws and mechanical tools, should be kept clean and maintained in
proper condition.
•Those parts that come into direct contact with the collected medicinal plant materials should be free from excess oil
and other contamination.

9. HARVESTING:
•Medicinal plants should be harvested during the optimal season or time period to ensure the production of medicinal
plant materials and finished herbal products of the best possible quality.
•Care should be taken to ensure that no foreign matter, weeds or toxic plants are mixed with the harvested medicinal
plant materials.
•Medicinal plants should be harvested under the best possible conditions, avoiding dew, rain or exceptionally high
humidity.
•If harvesting occurs in wet conditions, the harvested material should be transported immediately to an indoor drying
facility so as to prevent any possible deleterious effects due to increased moisture levels, which promote microbial
fermentation.
•Cutting devices, harvesters, and other machines should be kept clean and adjusted to reduce damage and
contamination from soil and other materials.
•If the underground parts (such as the roots) are used, any adhering soil should be removed from the medicinal plant
materials as soon as they are harvested.
•The harvested raw medicinal plant materials should be transported promptly in clean, dry conditions they may be
placed in clean baskets, dry sacks, trailers, hoppers or other well-aerated containers and carried to a central point for
transport to the processing facility.
•All containers used at harvest should be kept clean and free from contamination by previously harvested medicinal
plants and other foreign matter.
•When containers are not in use, they should be kept in dry conditions, in an area that is protected from insects, rodents,
birds and other pests, and domestic animals.
•Decomposed medicinal plant materials should be identified and discarded during harvest, post-harvest inspections and
processing, in order to avoid microbial contamination and loss of product quality.

10. The following general rules for collection of crude drugs:
• Roots and rhizomes are collected at the end of the vegetation period, i.e. usually in the autumn. In most cases they
must be washed free of adhering soil and sand.
• Bark is collected in the spring. Leaves and herbs are collected at the flowering stage.
• Flowers are usually gathered when fully developed.
• Fruits and seeds are collected when fully ripe.
Methods of collection:
• Medicinal plants must be largely collected by hand.
• With cultivation on a large scale, it may be possible to use modern agricultural harvesters, but in many cases, e.g.
barks, manual collection is unavoidable. Thus, the cost of drug production is largely the cost of the labor involved.
As per WHO Guidelines:
1. Medicinal plants/herbal drugs should be harvested when they are at the best possible quality for the proposed use.
2. Damaged plants or parts plants need to be excluded.
3. Medicinal plants/herbal drugs should be harvested under the best possible conditions avoiding wet soil, dew, rain
or exceptionally high air humidity. If harvesting occurs in wet conditions possible adverse effects on the medicinal
plant/herbal drug due to increased moisture levels should be counteracted.
4. Cutting devices or harvesters must be adjusted such that contamination from soil particles is reduced to a
minimum.
5. The harvested medicinal plant/herbal drug should not come into direct contact with the soil. It must be promptly
collected and transported in dry, clean conditions.

11. 6. During harvesting, care should be taken to ensure that no toxic weeds mix with harvested medicinal plants/herbal
drugs.
7. All containers used during harvesting must be clean and free of contamination from previous harvests. When
containers are not in use, they must be kept in dry conditions free of pests and inaccessible to mice/rodents, livestock and
domestic animals.
8. Mechanical damage and compacting of the harvested medicinal plant/herbal drug that would result in undesirable
quality changes must be avoided.
9. Freshly harvested medicinal plants/herbal drugs must be delivered as quickly as possible to the processing facility in
order to prevent thermal degradation.
10. The harvested crop must be protected from pests, mice/rodents, livestock and domestic animals. Any pest control
measures taken should be documented.
DRYING:
When medicinal plant materials are prepared for use in dry form, the moisture content of the material should be kept as
low as possible in order to reduce damage from mould and other microbial infestation. Medicinal plants can be dried in a
number of ways
1. In the open air (shaded from direct sunlight);
2. Placed in thin layers on drying frames, wire-screened rooms or buildings.
3. By direct sunlight, if appropriate.
4. In drying ovens/rooms and solar dryers.
5. By indirect fire; baking; lyophilization; microwave; or infrared devices.
6. Vacuum drying
7. Spray dryer: Examples: Papaya latex and pectin’s, etc.

12. The most common method for preserving plant material is drying.
Enzymatic processes take place in aqueous solution. Rapid removal of the water from the cell will, therefore, largely
prevent degradation of the cell constituents.
Drying also decreases the risk of external attack, e.g. by moulds.
leaves may contain 60-90% water, roots and rhizomes 70-85%,and wood 40-50%. The lowest percentage, often no
more than 5-10%, is found in seeds.
To stop the enzyme processes, the water content must be brought to about 10 %.
The plant material is spread out on shallow trays, which are placed on mobile racks and passed into a tunnel where
they meet a stream of warm air.
The air temperature is kept at 20-40 °C for thin materials such as leaves, but is often raised to 60-70 °C for plant parts
that are harder to dry, e.g. roots and barks.
When the crude drug has been collected under primitive conditions, without access to a drier, it must be dried in the
open.
STORAGE OF CRUDE DRUGS:
1.Storage facilities for medicinal material should be well aerated, dry and protected from light, and, when necessary, be
supplied with air-conditioning and Humidity control equipment as well as facilities to protect against rodents, insects.
2. The floor should be tidy, without cracks and easy to clean. Medicinal material should be stored on shelves which keep
the material a sufficient distance from the walls; measures should be taken to prevent the occurrence of pest infestation.
3. Continuous in-process quality control measures should be implemented to eliminate substandard materials,
contaminants and foreign matter prior to and during the final stages of packaging.
4. Processed medicinal plant materials should be packaged in clean, dry boxes, sacks, bags or other containers in
accordance with standard operating procedures and national and/or regional regulations of the producer and the end-
user countries.

13. 5. Materials used for packaging should be non-polluting, clean, dry and in undamaged condition and should conform to
the quality requirements for the medicinal plant materials concerned. Fragile medicinal plant materials should be
packaged in rigid containers.
6. Dried medicinal plants/herbal drugs, including essential oils, should be stored in a dry, well-aerated building, in which
daily temperature fluctuations are limited and good aeration is ensured.
7. Fresh medicinal plant materials should be stored at appropriate low temperatures, ideally at 2-8°C; frozen products
should be stored at less than -20°C.
8. Small quantity of crude drugs could be readily stored in air-tight, moisture proof and light proof container such as tin,
cans, covered metal tins or amber glass containers. 9. Wooden boxes and paper bags should not be used for storage of
crude drugs.
PRESERVATION OF PLANT MATERIAL :
The plant material must first be preserved so that the active compounds will remain unchanged during transport and
storage. The cells of living plants contain not only low molecular-weight compounds and enzymes, but they also have
many kinds of barriers that keep these constituents apart. When the plant dies, the barriers are quickly broken down
and the enzymes then get the opportunity to promote various chemical changes in the other cell constituents, e.g. by
oxidation or hydrolysis.

14. FACTORS INFLUCING THE CULTIVATION OF
MEDICINAL PLANTS

15. The following factors are influencing of cultivation:
1. Light:
• Light is the only external source of energy for the continuation of life of the plant. It influences photosynthesis,
opening and closing of stomata, plant movements, seed germination, flowering and vegetative growth like tuber
formation.
• Dry sunny weather increases the proportion of glycosides in digitalis and of alkaloids in belladonna.
2. Temperature:
• Temperature is the major factor influencing the cultivation of the medicinal plant.
• The sudden decrease in temperature caused the formation of the ice crystals in intercellular spaces of the plant.
• As a result, water comes out of the cells and ultimately plants die due to drought and desiccation.
• The ice crystals also mechanical injury to the cells temperature stimulates the growth of seedlings.
• Water absorption decreases at low temperatures. The rate of photosynthesis is affected by change in temperature.
• The rate of respiration increases with increase in temperature. Examples; Cinchona- 58-73°F; Tea- 75- 90°F and
coffee- 55-70°F.
3. Atmosphere humidity:
• It is present in the form of water vapours. This is called atmospheric humidity.
• Clouds and fog are the visible forms of humidity.
• The major sources of water vapours in the atmosphere are evaporation of water from earth surface and
transpiration from plants the major effect of humidity on plant life and climate.
• Evaporation of water, its condensation and precipitation depends upon relative humidity and humidity affects
structure, form and transpiration in plants.

16. 4. Altitude:
• The altitude is the most important factor influencing of cultivation of medicinal plants.
• The increase the altitude, the temperature and atmospheric pressure decreases while the wind velocity, relative
humidity and light intensity increases.
• Thus, as the climatic conditions change with height, they also produce change in the vegetation pattern. The bitter
constituents of Gentiana lutea increase with altitude, whereas the alkaloids of Aconitum nacelles and lobelia inflate and
oil content of thyme and peppermint decrease.
• Pyrethrum gives the best yield and Pyrethrum at high altitude. Examples: Tea- 9500-1500 meters; cinnamon- 300-
1000 meters and saffron- up to 1250 meters.
5. Rainfall:
• The rainfalls are most important factor influencing of cultivation of medicinal plants.
• The main source of water for the soil is rain water. Rainfall and snowfall have a large effect the climate condition.
• The water from rainfall flows into the rivers and lakes percolates into the soil to form ground water and remaining is
evaporated.
• The minerals in the soil get dissolved in water and are then absorbed by plants.
• Water influences morphological and physiology of plant.
• Examples: continuous rain can lead to a loss of water- soluble substance from leaves and root by leaching; this is
known to apply to some plants producing glycoside and alkaloids.

17. 6. Soil:
• Soil is defined as surface layer of the earth, formed by weathering of rocks.
• The soil is formed as a result of combined action of climate factors like plants and microorganisms.
• The soil should contain appropriate amounts of nutrients, organic matter and other elements to ensure optimal
medicinal plant growth and quality.
• Optimal soil conditions, including soil type, drainage, moisture retention, fertility and pH, will be dictated by the selected
medicinal plant species and/or target medicinal plant part.
The soil made of five components:
(i) Mineral matter. (ii) Soil air. (iii) Soil water. (iv) Organic matter or humus. (v) Soil organisms
Plants depend on soil for nutrients, water supply and anchorage.
Soil influences seed germination, capacity of plant to remain erect, form, vigour and woodiness of the stem, depth of root
system, number of flowers on a plant, drought, frost, etc.
Fertilizer:
• 1. Biological origin fertilizer. • 2. Synthetic fertilizers • 3. Chemical fertilizer
1. Biological origin fertilizer:
• Soil is generally poor in organic matter and nitrogen.
• The substances of biological origin used as fertilizer are thus selected if these could provide the elements required.

18. These are two types:
(i) Green manures:
• Manure is material, which are mixed with soil.
• These supply almost all the nutrients required by the crop plants.
• This results in the increase in crop productivity.
(ii) Bio-fertilizer:
• It can be defined as biologically active products or bacteria, algae and fungi which useful in bringing about soil nutrient
enrichment.
• These mostly include nitrogen fixing microorganisms.
2. Synthetic fertilizer:
• Are “Man made” inorganic compounds - usually derived from by-products of the petroleum industry.
• Examples are Ammonium Nitrate, Ammonium Phosphate, Superphosphate, and Potassium Sulfate. Plants require 13
nutrients.
3. Chemical fertilizers:
(i) Macronutrients: • (a) Nitrogen • (b) Phosphorous • (c) Potassium • (d) Calcium • (e) Magnesium • (f) Sulpher.
(ii) (ii) Micronutrients: • (a) Iron • (b) Magnese • (c) Zinc • (d) Boron • (e) Copper • (f) Molybdenum Examples: • Urea,
Potash

19. PLANT HORMONES AND THEIR APPLICATIONS

20. Plant Growth Regulators (Hormones):
• The organic compounds, other than nutrients, which effect the morphological structure and physiological processes of
plants in low concentrations are known Plant Growth Regulators or Phyto hormones or plant hormones.
• Plant hormones control the complete plant lifecycle, including germination, rooting, growth, flowering, fruit ripening,
foliage and death.
• They induced native and synthetic action on plant growth.
Plant growth regulators are as: 1. Auxins 2. Gibberellins 3. Cytokinins 4. Abscisic acid 5. Ethylene.
1. Auxins:
• Auxins were the first plant hormones discovered.
• Auxin is a general term used to indicate substances that promote elongation of tissues. • Indole acetic acid (IAA) is an
auxin that occurs naturally in plants.
• Natural auxins – • Indole -3- acetonitrile (IAN) • Phenyl acetic acid • Synthetic auxins - • Indole -3- Butyric Acid(IBA), •
α-Naphthyl Acetic Acid (NAA), • 1-naphthyl acetamide (NAD)
• Functions of auxin:
• Stimulates internode elongation.
• Stimulates leaf growth.
• Stimulates initiation of vascular tissue, fruit growth.
• Inhibition of root growth.

21. • Differentiation of vascular tissue (xylem and phloem) is stimulated by IAA.
• Auxin stimulates root initiation on stem cuttings.
• Stimulates lateral root development in tissue culture (adventitious rooting).
• Auxin mediates the tropic response of bending to gravity and light.
2. Gibberellins:
• gibberellins occur in green plants, fungi and bacteria.
• According to a research carried out in Japan, USA and Britain has shown that Gibberellins A – isolated in 1938 – is
actually a mixture of at least 6 gibberellins named as – GA1, GA2, GA3, GA4, GA7, and GA9 • The gibberellins are named
GA. GAn in order of discovery.
• GA3 is termed as Gibberellic acid.
• There are currently 50 GAs identified from plants, fungi and bacteria.
• 40 of these occur in green plants.
Functions of Gibberellins:
• Stimulates stem elongation by stimulating cell division and elongation.
• GA controls internode elongation in the mature regions of plants.
• Dwarf plants do not make enough active forms of GA.
• Flowering in biennial plants is controlled by GA. Biennials grow one year as a rosette and after the winter, they bolt
(rapid expansion of internodes and formation of flowers)
• Breaks seed dormancy in some plants.

22. • Stimulates α-amylase and other hydrolytic enzymes during germination of monocot seeds.
• Stimulates germination of pollen and growth of pollen tubes.
• Can cause parthenocarpic (seedless) fruit development or increase the size of seedless fruit (grapes).
3. Cytokinins:
• Cytokinins are compounds with a structure resembling adenine.
• Cytokinin have been found in almost all higher plants as well as mosses, fungi, bacteria, and also in many prokaryotes
and eukaryotes.
• There are more than 200 natural and synthetic cytokinins identified.
• The first naturally occurring cytokinin was isolated from corn in 1961 by Miller and it was later called zeatin.
• The naturally occurring cytokinins are zeatin, N6 dimethyl amino purine, isopentenyl aminopurine.
• The synthetic cytokinins are kineatin, adenine, 6-benzyl adenine benzimidazole and N, N’-diphenyl urea.
Functions of Cytokinins:
• Stimulate cell division (cytokinesis).
• Stimulate morphogenesis (shoot initiation/bud formation) in tissue culture.
• Stimulate the growth of lateral (or adventitious) roots.
• Stimulate leaf expansion resulting from cell enlargement.
• May enhance stomatal opening in some species.
• Promotes the conversion of etioplasts into chloroplasts.
• Promotes some stages of root development.

23. 4. Ethylene:
• It is a simple organic molecule present in the form of volatile gas - in ripening fruits, flowers, stem, roots, tubers, seeds.
• It is present in very small quantity, but its quantity increases during the time of growth and development.
• Ethylene is responsible for • fruit ripening, • leaf abscission, • stem swelling, • leaf bending, • flower petal discoloration
and • inhibition of stem and root growth,
• It is commercially used for promotion of flowering and fruit ripening, and stimulation of latex flow in rubber trees.
5. Abscisic acid (ABA):
• The physiological activities in plants like retaining or shedding of different organs such as leaves, flowers and fruits
requires a natural growth inhibitor.
• Other synthetic ABA are Maleic hydrazide, Daminozide, Glyphosine, Chlorophonium chloride.
• It inhibits the gibberellins induced synthesis of amylase and other hydrolytic enzymes.
• ABA accumulates in many seeds and helps in seed dormancy.
• ABA serves as potential anti-transpirent by closing the stomata, when applied to leaves.

24. POLYPLOIDY, MUTATION AND HYBRIDIZATION
WITH REFERENCE TO MEDICINAL PLANTS

25. WHAT IS POLYPLOIDS ?
• Polyploids are organisms with multiple sets of chromosomes in excess of the diploid number .
• Polyploidy is common in nature and provides a major mechanism for adaptation and speciation.
• Approximately 50-70% of angiosperms, which include many crop plants, have undergone polyploidy during their
evolutionary process.
CLASSIFICATION OF POLYPLOIDS:
Based on their chromosomal composition: euploids aneuploids.
• Euploids constitute the majority of polyploids
EUPLOI DY:
• are polyploids with multiples of the complete set of chromosomes specific to a species. • Depending on the composition
of the genome, euploids can be further classified into autopolyploi ds allopolyploid.
AUTOPOLYPLOIDY:
• Containing ofmultiple copies of the basic set (x) of chromosomes of the same genome.
• occurs in nature through union of unreduced gametes.
• Natural autoploids include tetraploid crops such as alfafa, peanut, potato and coffee and triploid bananas.

26. ALLOPOLYPLOIDY:
• A combination of genomes from different species .
• They result from hybridization of two or more genomes followed by chromosome doubling or by the fusion of
unreduced gametes between species .
• This mechanism is called non-disjunction . These meiotic aberrances result in plants with reduced vigor.
• Economically important natural alloploid crops include strawberry, wheat, oat, upland cotton, oilseed rape, blueberry
and mustard.
ANEUPLOIDY:
• are polyploids that contain either an addition or subtraction of one or more specific chromosome(s) to the total
number of chromosomes that usually make up the ploidy of a species.
• Aneuploids result from the formation of univalents and multivalents during meiosis of euploids.
• With no mechanism of dividing univalents equally among daughter cells during anaphase I, some cells inherit more
genetic material than others.
• Similarly, multivalents such as homologous chromosomes may fail to separate during meiosis leading to unequal
migration of chromosomes to opposite poles.

27. INDUCING POLYPLOIDS:
• They occur spontaneously through the process of chromosome doubling.
• Spontaneous chromosome doubling in ornamentals and forage grasses has led to increased vigour.
• Examples tulip forage grasses ryegrasses have yielded superior varieties following spontaneous chromosome
doubling.
Breeders have harnessed the process of chromosome doubling in vitro through induced polyploidy to produce superior
crops.
• For example, induced autotetraploids in the watermelon crop are used for the production of seedless triploid hybrids
fruits.
• Such polyploids are induced through the treatment of diploids with mitotic inhibitors such as dinitroaniles and
colchicine.
• It is necessary to eliminate duplicated genes in a newly formed polyploid to avoid gene silencing as well as to stabilize
fertility
• The increase in nuclear ploidy affects the structural and anatomical characteristics of the plant.
• Polyploidy results in increased leaf and flower size , stomatal density, cell size and chloroplast count.
• Hybrid vigor resulting from interspecific crosses in allopolyploids is one of the most exploited advantages of polyploid
in plant breeding.
A comparison between the leaf and flower of a (A) diploid and (B) induced tetraploid watermelon A B

28. MUTATION WITH REFERENCE TO MEDICINAL PLANTS:
INTRODUCTION:
• Sudden heritable change in genetic material or character of an organismis known as mutation • Individuals showing
these changes are known as mutants.
• An individual showing an altered phenotype due to mutation are knownas variant • Factor or agents causing mutation
are known as mutagens • Mutation which causes changes in base sequence of a gene are knownas gene mutation or point
mutation.
HISTORY:
• English farmer Seth Wright recorded case of mutation first time in 1791 in male lamb with unusual short legs.
• The term mutation is coined by Hugo de Vries in 1900 by his observation in Oenothera.
• Systematic study of mutation was started in 1910 when Morgan genetically analyzed white eye mutant of Drosophila.
• H. J. Muller induced mutation in Drosophila by using X- rays in 1927 ; he was awarded with Nobel prize in 1946
CHARACTERISTICS OF MUTATION:
• Generally mutant alleles are recessive to their wild type or normal alleles.
• Most mutations have harmful effect, but some mutations are beneficial.
• Spontaneous mutations occurs at very low rate.
• Some genes shows high rate of mutation such genes are called as mutable gene.
• Highly mutable sites within a gene are known as hotspots.
• Mutation can occur in any tissue/cell (somatic or germinal) of an organism.

29. CLASSIFICATION OF MUTATION:
• Based on the survival of an individual:
1. Lethal mutation – when mutation causes death of all individuals undergoing mutation are known as lethal.
2. Sub lethal mutation - causes death of 90% individuals.
3. Sub vital mutation– such mutation kills less than 90% individuals.
4. Vital mutation -when mutation don’t affect the survival of an individual are known as vital.
5. Supervital mutation – This kind of mutation enhances the survival of individual.
• Based on causes of mutation:
1. Spontaneous mutation- Spontaneous mutation occurs naturally without any cause. The rate of spontaneous
mutation is very slow eg- Methylation followed by deamination of cytosine. Rate of spontaneous mutation is higher in
eukaryotes than prokaryotes. Eg. UV light of sunlight causing mutation in bacteria
2. Induced Mutation- Mutations produced due to treatment with either a chemical or physical agent are called induced
mutation . The agents capable of inducing such mutations are known as mutagen. use of induced mutation for crop
improvement program is known as mutation breeding. Eg. X- rays causing mutation in cereals
• Based on tissue of origin:
1. Somatic mutation- A mutation occurring in somatic cell is called somatic mutation. In asexually reproducing species
somatic mutations transmits progeny to the next progeny.
2. Germinal Mutation- from one When mutation occur in gametic cells or reproductive cells are known as germinal
mutation. In sexually reproductive species only germinal mutation are transmitted to the next generation.

30. • Based on direction of mutation:
1.Forward mutation- When mutation occurs from the normal/wild type allele to mutant allele are known as forward
mutation.
2.Reverse mutation- When mutation occurs in reverse direction that is from mutant allele to the normal/wild type
allele are known as reverse mutation.
• Type of trait affected:
1. Visible mutation- affects on phenotypic character Those mutation and can be which detected by normal
observation are known as visible mutation.
2. Biochemical mutation- mutation which affect the production of biochemicals and which does not not show any
phenotypic character are known as biochemical mutation.
CHROMOSOME MUTATIONS:
• May Involve: – Changing the structure loss or gain
CHROMOSOME MUTATIONS :
• Five types exist: –Deletion –Inversion –Translocation – Nondisjunction –Duplication
1. DELETI ON
• Due to breakage • A piece of a chromosome is lost.
2. INVERSI ON
• Chromosome segment breaks off • Segment flips around backwards • Segment reattaches

31. 3. DUPLICAT ION
• Occurs when a gene sequence is repeated.
4. TRANSLOCA TION
• Involves two chromosomes that aren’t homologous • Part of one chromosome is transferred to another
chromosomes.
5. NONDISJUNC TION
• Failure of chromosomesto separate during meiosis
• Causes gamete to have too many or too few chromosomes
• Disorders: – Down Syndrome – – Turner Syndrome – – Klinefelter’s Syndrome –
TYPES OF GENE MUTATIONS:
• –Point Mutations • –Substitutions • –Insertions • –Deletions • –Frameshift
• POINT MUTATION • Change of a single nucleotide • Includes the deletion, insertion, or substitution of ONE nucleotide
in a gene
POINT MUTATION • Sickle Cell disease is the result of one nucleotide substitution • Occurs in the hemoglobin gene
• FRAMESHIFT MUTATION • Inserting or deleting one or more nucleotides • Changes the “reading frame” like
changing a sentence • Proteins built incorrectly

32. HYBRIDIZATION WITH REFERENCE TO MEDICINAL PLANTS:
MEANING OF HYBRIDIZATION:
• Individual produced as a result of cross between two genetically different parents is known as hybrid. The natural or
artificial process that results in the formation of hybrid is known as hybridization.
• The production of a hybrid by crossing two individuals of unlike genetical constitution is known as hybridization.
Hybridization is an important method of combining characters of different plants. Hybridization does not change
genetic contents of organisms but it produces new combination of genes.
OBJECTIVES OF HYBRIDIZATION:
1. To artificially create a variable population for the selection of types with desired combination of characters.
2. To combine the desired characters into a single individual.
3. To exploit and utilize the hybrid varieties.
TYPES OF HYBRIDIZATION:
(i) Intra-varietal hybridization: The crosses are made between the plants of the same variety.
(ii) Inter-varietal or Intraspecific hybridization: The crosses are made between the plants belonging to two different
varieties.
(iii) Interspecific hybridization or intragenric hybridization: The crosses are made between two different species of the
same genus.

33. Procedure of Hybridization:
It involves the following steps:
(i) Selection of parents.
(ii) Selfing of parents or artificial self-pollinat
(iii) Emasculation.
(iv) Bagging
(v) Tagging
(vi) Crossing
(vii) Harvesting and storing the F, seeds
(viii) Raising the F1 generation.

34. CONSERVATION OF MEDICINAL PLANTS

35. Medicinal plants and traditional medicine play an important role in the health care system of most developing countries.
The traditional health care practice is mainly dependent on medicinal plants collected from the wild. In spite of this, the
medicinal plant biodiversity is being depleted due to man-made and natural calamities. Moreover, the indigenous
knowledge associated with the conservation and use of medicinal plants is also disappearing at an alarming rate. The
fact that medicinal plants could be used as sources of revenue for farmers, the Institute of Biodiversity Conservation
(IBC) has initiated the development of a project on Conservation and Sustainable Use of Medicinal Plants (CSMPP).
NEED FOR CONSERVATION:
• The goal of conservation is to support sustainable development by protecting and using biological resources in ways
that do not diminish the world’s variety of genes and species or destroy important habitats and ecosystems.
• In general, it involves activities such as collection, propagation, characterization, evaluation, disease indexing and
elimination, storage and distribution.
• The conservation of plant genetic resources has long been realised as an integral part of biodiversity conservation.
• There are two methods for the conservation of plant genetic resources, namely In- Situ & Ex-Situ conservation.
• On the other hand, ex situ conservation involves conservation outside the native habitat and is generally used to
safeguard populations in danger of destruction, replacement or deterioration

36. GERMPLASM TECHNIQUE FOR CONSERVATION:
• Germplasm conservation of vegetatively propagated crops, forest species especially those with recalcitrant seeds in
live gene banks in fields poses tremendous problems in terms of required land space and labour input during annual
or perennial replanting, testing and documentation.
• The advantage of in vitro or reduced growth storage include little space necessary in growth rooms for maintaining
thousands of genotypes and the absence of diseases and pest attack in culture vessels.
• Furthermore, in vitro storage eliminates the need for long and frustrating quarantine procedures during movement
and exchange of germplasm
Disadvantages of Germplasm:
• Some crops do not produce viable seeds.
• Some seeds remain viable for a limited duration only and are recalcitrant to storage Seeds of certain species
deteriorate rapidly due to seed borne pathogen. Some seeds are very heterozygous not suitable for maintaining true
to type genotypes.

37. CRYOPRESERVATION TECHNIQUE FOR CONSERVATION OF PLANTS:
“Cryopreservation” is defined as the viable freezing of biological material and their subsequent storage at ultra low
temperatures (- 196C)”using liquid nitrogen.
The use of liquid nitrogen, either by itself or as a source of nitrogen gas, is based on the following unique combination of
features:
• Chemically inert • Relatively low cost • Non-toxic • Non-flammable • Readily available Factors determining the
Conservation Protocol • Applicability. • Range, time, reproductive biology, storage suitability, infrastructure. • Need
Driven and Technology driven • Security, Efficiency, Accessibility and Sustainability. • Resource requirement. • Risks in
conservation • Regeneration capability and time. • Cost and returns.
New cryopreservation techniques:
Encapsulation and dehydration, Vitrification, Encapsulation and vitrification, Desiccation, Pregrowth, Pregrowth and
desiccation, Droplet freezing.
Cryopreservation procedures:
Three different procedures have been used for cryopreservation of plant cells: two-step freezing, vitrification and
encapsulation dehydration.

38. Two-step freezing:
This procedure includes an incubation of cells in a mixture of cryoprotectants (total concentration of 1–2M), which
causes moderate dehydration of the cells, followed by a slow freezing step (for example, 1°C/min down to app –35°C).
Vitrification:
This procedure is based on severe dehydration at non-freezing temperatures by direct exposure to concentrated
cryoprotectants (total concentration ranging from 5–8M), followed by rapid freezing.
Encapsulation-dehydration:
Cells are encapsulated in alginate beads, cultured on medium with increased sucrose concentration, air-dried using
silica gel or the airflow of a flow cabinet and directly transferred to liquid nitrogen.