CULTIVATION, COLLECTION, PROCESSING AND STORAGE OF CRUDE DRUGAS.pptx

Pharmacognosy & Phytochemistry -I
CULTIVATION, COLLECTION, PROCESSING AND STORAGE OF CRUDE DRUGS

1. CULTIVATION, COLLECTION, PROCESSING AND STORAGE OF CRUDE DRUGS Prepared by: Ms. Ithape G.C. ( M.pharm )

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.

3. ADVANTAGES OF CULTIVATION It ensures quality and purity of medicinal plants.Collection of crude drugs from cultivated plants gives a better yield and therapeutic quality. Cultivation ensures regular supply of a crude drug. The cultivation of medicinal and aromatic plants also leads to industrialization to a greater extent. Cultivation permits application of modern technological aspects such as mutation, polyploidy and hybridization.

4. DISADVANTAGES OF CULTIVATION The high cost of cultivation drugs as compared to wild source• Losses due to ecological imbalance such as storms, earthquakes, floods, droughts etc Methods of Propagation Sexual Method (Seed Propagation) Asexual Method (Vegetative Propagation)

5. Asexual Method (Vegetative Propagation) Vegetative propagation can be defined as regeneration or formation of a new indivisual 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. They are two types of Vegetative propagation: A. Natural Methods of vegetative propagation B. Artificial Methods of vegetative propagation

6. Vegetative propagation Vegetative propagation can be defined asregeneration or formation of a new individual from any vegetative part of the plant body. A.Natural Methods of vegetative propagation The method of vegetative propagation involves separation of a part of plant body, which develops into a new plant Bulb Allium, Squill Corms Colchicum Tubers Potato, aconite Rhizomes Ginger and turmeric Suckers Pineapple, banana

7. B. Artificial Methods of vegetative propagation The method by which plantlets or seedlings are produced from vegetative part by using some technique or process is known as artificial method of vegetative propagation. Following methods are used: Cutting Layering Grafting Micro propagation

8. 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. 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. 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.

9. 4. Micro propagation: 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

10. Importance of asexual propagation 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. 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. All the plants developed by these methods will be generally similar to the parent plant. Micro propagation is useful in raising disease free plants.

11. 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.

12. 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

13. Sexual Method (Seed Propagation) Incase of sexual reproduction, the plants are raised from seeds and such plants Pollination: This is the transfer of pollen grains from the anther to the stigma. Fertilization: Fusion of male and female gametes takes place, resulting in the formation of zygote Seedling: Multiplication of plants by using seed is called as seed propagation. METHODS OF SOWING SEEDS Numerous methods of sowing seeds are in practice. BROADCASTING DIBBLING MISCELLANEOUS

14. BROADCASTING In this method the seeds are scattered freely in well prepared soil for cultivation. The seeds only need raking. If they are sown deep or covered by soil, necessary thinning of the seedlings is done by keeping a specific distance Examples: isapgol linseed, sesame DIBBLING When the seeds of average size and weight are available. They are sown by placing in holes. Number of seeds to be put in holes vary from 3 to 5 Depending upon the vitality

15. MISCELLANEOUS Many a times the seeds are sown in nursery beds. The seedling thus produced are transplanted to farms for further growth. Such as cinchona, cardamom, clove, etc.

16. Pre-germination Seed Treatments to improve germination rate 1. Chemical (Acid scarification) 2. Mechanical (Scarification) 3. Boiled Water Treatment 4. Soaking in Water 5. Moist Chilling

17. Chemical (Acid scarification) The purpose is to modify hard or impermeable seed covering by soaking seed in concentrated sulphuric acid The time of treatment may vary from 10 minutes to 6 hour according to speciesON 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 Placing the seeds between two sand paper Passing seeds through machine that scratches the surface. Filling and notching to make the seed coat permeable to water

18. 3. 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 growth 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

19. 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, tornato . 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

20. 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.

21. 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.

22. 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, shouldbe 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

23. 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

24. 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 duringharvest , post-harvest inspections and processing, in order to avoid microbial contamination and loss of product quality.

25. 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

26. As per WHO Guidelines Medicinal plants/herbal drugs should be harvested when they are at the best possible quality for the proposed use. Damaged plants or parts plants need to be excluded. 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. Cutting devices or harvesters must be adjusted such that contamination from soil particles is reduced to a minimum. 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.

27. 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.

28. 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 In the open air (shaded from direct sunlight); Placed in thin layers on drying frames, wire-screened rooms or buildings. By direct sunlight, if appropriate. In drying ovens/rooms and solar dryers. By indirect fire; baking; lyophilization ; microwave; or infrared devices. Vacuum drying Spray dryer: Examples: Papaya latex and pectin's, etc.

29. 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

30. 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.

31. 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.

32. 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.

33. FACTORS INFLUCING THE CULTIVATION OF MEDICINAL PLANT Light Temperature Atmosphere humidity Altitude Rainfall Soil Fertilizer Plant hormone Polyploidy Hybridization Green house effect

34. Students have to eloborate all those factor

35. 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.

36. 2. Temperature: Temperature is the major factor influencin 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

37. 3. Atmosphere humidity: It is present in the form of water vapours . Tuis 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.

38. 4. Altitude: The altitude is the most important factor influencii . 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

39. Plant Altitude ( metres ) Altitude ( Metre ) Tea 1,000-1,500 Cinchona 1,000-2,000 Camphor 1,500-2,000 Cinnamon 250-1,000 Coffee 1,000-2,000 Clove Up to 900 Saffron Up to 1,250 Cardamom 600-1,600

40. 5. Rainfall: The rainfalls are most important factor influencir 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.

41. Crops Water requirement to produce 1kg (in liters) Rice 2497 Potato 287 Tomato 214 Cucumber 235 Cabbage/Lettuce 237 Maize/Corn 1222 Wheat 1350

42. 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.

43. The soil made of five component Mineral matter. Soil air. Soil water. Organic matter or humus. 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

44. 7. Fertilizer: Biological origin fertilizer. Synthetic fertilizers Chemical fertilizer 1 . Biological origin fertilizer. Mannures : Manure is materials which are mixed with soil. Thses supply almost all the nutrients required by the crop plants. This results in the increases in crop productivity. Munnures are three types: 1.Farmyard manure-FYM (Cow dung manure, Poultry manure.) 2.Composited Manure-(Organic nitrogen supplements, Bone meal, Fish meal.) 3.Green manure- Neem seed cake, Vermi compost, Oil cake. Bio fertilizer : Can be defined as biologically active products or bacteria, algae and fungi, which useful in bringing about soil nutrient enrichment. e.g : Rhizobium,Azobactor Major Nutrients: Nitrogen (N), Phosphorus (P), and Potassium (K) Secondary Nutrients: Calcium (Ca), Magnesium (Mg), and Sulfur (S) Micronutrients or Trace Elements: Boron (B), Chlorine (CI), Copper (Cu), Iron (Fe),Manganese (Mn), Molybdenum (Mo), and Zinc (Zn).

45. 2. Chemical fertilizers: Macronutrients: (a) Nitrogen(b) Phosphorous(c) Potassium(d) Calcium(e) Magnesium(f) Sulpher . (ii) Micronutrients: (a) Iron(b) Magnese (c) Zinc(d) Boron(e) Copper(f) Molybdenum Examples:Urea , Potash

46. 3. 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. 16 Elements Necessary for Plants growth and developments . Carbon (C) 2. Hydrogen (H) 3. Oxygen (0) 4. Nitrogen (N) 5.Phosphorus (P) 6. Potassium (K) 7. Calcium (Ca) 8. Magnesium (Mg) 9. Sulfur (S) 10. Boron (B) 11. Chlorine (CI) 12. Copper (Cu) 13. Iron (Fe) 14. Manganese ( Mn ) 15. Molybdenum (Mo) 16. Zinc (Zn)

47. 8.HORMONES It is a hormone like synthetic organic compound. In small amounts, it mouns ute growth and development either by promoting or inhibiting the growth. General plant hormones:The phyto -hormones are broadly grouped under five major classes namely Auxins (cell elongation) Gibberellins (cell elongation + cell division - translated into growth) Cytokinins (cell division + inhibits senescence) Abscisic acid (abscission of leaves and fruits) Ethylene (promotes senescence, epinasty , and fruit ripening)

48. 9.POLYPLOIDY The term euploidy is a type of ploidy in which genome contain whole set of chrome euploidy includes monoploidy , diploidy and polyploidy. When some plants contain more than two genomes it is called as polyploidy. Polyploidy is caused by artificially induced methods/physical agents Colchicine (b) Veratrine (C) Sulphanilamide (d) Mercuric chloride. Significant Effects of polyploidy : Greater significance to medicinal plants. It may cause formation of new species. Mutation: Sudden change in genotype causing qualitative or quantitative alteration of genetic material called mutation. Mutation can artificially produced by certain agents called mutagenes Types of mutagenes : (A) Physical Mutagenes : Ionizing radiations: x-rays, gamma radiation and cosmic rays. Non Ionizing radiations: UV radiation ( B) Chemical Mutagenes : Alkylating agents: Nitrogen and sulphur mustard. Acridines : Acridines and proflavins . Nitrous acid.

49. 10.HYBRIDIZATION Hybridization is the process of crossing two genetically different individuals to result in a third individual with a different, often preferred, set of traits. Plants of the same species cross easily and produce fertile progeny. Wide crosses are difficult to make and generally produce sterile progeny because of chromosome-pairing difficulties during meiosis. Hybridization of plants occurs in nature through various mechanisms. Some plants (such as the oil palm) are insect-pollinated, and others (such as maize, or corn) are wind-pollinated. Such plants are referred to as cross-pollinated plants. Natural hybridization has played a significant role in producing new genetic combinations and is the norm in cross-pollinated plants. It is a common way of generating genetic variability.

50. 11.GREEN HOUSE EFFECT There is a "natural" greenhouse effect that keeps the Earth's climate habitable. Normally a sun ray reaches the earth and heat is radiated back in to space. However, when Co, concentration increases in the atmosphere, it forms a thick cover and prevents the heat from being re-radiated. Consequently, the atmosphere gets heated and the temperature increases.This is called GREEN HOUSE EFFECT. There is also the "man-made" green house effect, which is the enhancement of Earth's natural greenhouse effect by the addition of greenhouse gases from the burning of fossil fuels (mainly petroleum, coal, and natural gas). Greenhouse gases include water vapour , carbon dioxide, methane, nitrous oxide, ozone and some artificial chemicals such as chlorofluorocarbons (CFCs). This mainly effects plant growth, development and yield.

51. 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: Auxins Gibberellins Cytokinins Abscisic acid Ethylene.

52. 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), a-Naphthyl Acetic Acid (NAA), 1-naphthyl acetamide (NAD)

53. Functions of auxin Stimulates internode elongation. Stimulates leaf growth. Stimulates initiation of vascular tissue, fruit growth. Inhibition of root growth. 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.

54. 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,GA5,, and GA6 GA, is termed as Gibberellic acid. There are currently 50 GAs identified from plants, fungi and bacteria. 40 of these occur in green plants.

55. 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. Stimulates a-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).

56. 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, Nº dimethyl amino purine, isopentenyl aminopurine. The synthetic cytokinins are kineatin , adenine, 6-benzyl adenine benzimidazole and N, N'-diphenyl urea.

57. 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.

58. 4. Ethylene : It is a simple organic molecule present in the form of volatile gas in ripening fruits, flowers, stem, roots, tubers, seeds. 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.

59. 5. Abscisic acid (ABA): The physiological activities in plants like retaining or shedding 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.

60. Polyploidy reference to medicinal plants •Polyploidy word is the combination of two words- Poly"+ „ Ploidy Poly means many or more and Ploidy means the number of pairs or sets of chromosomes available in cell of any living things. So polyploidy defined as the condition at which the number of sets or pairs of chromosomes will be more than two in cell of any living things. In which one set inherited from each parents or excess of the diploid number

61. Haploid (In= gametes or chromatids ) diploid (2n) are normal form of chromosomes triploid (3n) & tetraploid (4n) are the example of polyploidy

62. Types Of Polyploidy They are two types as follows:- Euploidy Autopolyploidy b) Allopolyploidy B . Aneuploidy

63. Euploidy : - Itis the types of polyploidy with multiple numbers of sets of chromosomes which combination of the genomes. It is further classified in to 2 types: Autopolyploidy :- Auto means self or same species. Polyploidy"means the multiplication is complete in numbers to a specific species depending upor the number of chromosomes. So the multiplication the number of chromosomes between in same species is called " autopolyploidy ".. Allopolyploidy :- Allo means other or different species. Polyploidy means the multiplication the number of chromosomes. So the multiplication the number of chromosomes between in different species is called " allopolyploidy "

64. B. Aneuploidy : Aneuploids are polyploids that contain either an addition or subtraction of or more specific chromosome(s) to the total number of chromosomes that usually make up the ploidy of a species. Term Chromosome Number Monosomy 2n-1 Nullisomy 2n-2 Trisomy 2n+1 Tetrasomy 2n+2 Pentusomy 2n+3

65. Causes Of Polyploidy There are four types of causes behind the polyploidy:- Non-disjunction in mitosis Non-reduction in meiosis Polyspermy Endo-replication or Endo-reduplication

66. Factors Promoting Polyploidy There are three factors which promotes the polyploidy which are as follows: Physical factor Chemical factor Biological factor

67. Physical factor:- Temperature:- heat temperature & cold temperature Centrifugation X-rays Gamma rays Cosmic rays Ionizing & non-ionizing radiations UV-radiation

68. Chemical factor:- Alkylating agents:- nitrogen & sulphur mustard Acridines Proflavins Nitrous acid Colchicines VI. Colchicines (Poisonous alkaloids):-

69. 3. Biological factor Mode of reproduction Mode of fertilization Breeding system present (Hybridization) Growth habit of the plant Size of chromosomes

70. Application Of Polyploidy Mutation breeding Seedless fruits production Bridge crossing Ornamental & forage breeding Disease resistance through aneuploidy Industrial application of polyploidy Enlargement & increased vigour which is strength of the plant Production of sterile triploids seedless fruits, generally by the help of vegetative propagation except seedling & fertilization. Restoring fertility in hybrids . Overcoming barriers in hybridization . Enhancing pest resistance, disease resistance &stress tolerance plant

71. Mutation reference to medicinal plants• Sudden heritable change in the structure of a gene on chromosome or change the chromosome number. Type of mutations : Spontaneous and induced mutations . Recessive and dominant mutations . Somatic and germinal mutations . Forward , back and suppressor mutation . Chromosomal , genomic and point mutations

72. Mutations can be artificially produced by certain agents called mutagens or mutagenic agent. They are two types: Physical mutagens: Ionizing radiations: X-rays, gamma radiation and cosmic rays. Non-ionizing radiation: U.V. radiation,

73. b. Chemical mutagens : Alkylating and hydroxylating agents: Nitrogen and sulpher mustard; methyl a ethylsulphonate , ethylethane sulphonates . Nitrous acid : Acridines : Acridines and proflavins .

74. Application Of Mutation: This method is rapid method and cheap for development of new variety. Induced mutagen are used for the induction of CMS ( Congential Myasthenic Syndrome)Example. Ethidium bromide is used for induction of CMS in barley. It is effective for improvement of oligogenic character. This method is quik , simple and best way when a new character is to be induced. It improves the disease resistance in crop plant. It is used to improve the specific characters of well adapted high yielding varities .

75. Hybridization reference to medicinal plants It is mating or crossing of two genetically dissimilar plants having desired genes or genotypes and bringing them together into one individual called hybrid. The process through which hybrids are produced is called hybridization . A hybrid is an organism which results from crossing of two species or varieties differing at least in one set of characters.

76. Based on the nature & relationship of plants to be crossed, hybridization can be Inter-varietal - Cross bet. plants of two diff. var. of samespecies - intra- specific hybridization. Eg . Hybrid Maize. Intra-varietal - Cross bet. two plants of diff. genotypes butsame variety. Inter-specific Cross bet. two species of genus - Eg . Wheat, Cotton, Tobacco. Inter-generic- Cross bet. two diff. genera. Eg . Sugarcane X Bamboo, Wheat X Rye, Radish X Cabbage.

77. The following steps are involved in hybridization of plant : Choice Of Parents Selfing Of Parents Emasculation Bagging Crossing Or Cross Pollination Labelling Collection Of Hybrid Seeds

78. 1. Choice of parents: First step in hybridization. Desirable male & female plants are selected. Parent plants should be healthy & vigorous. Parents to be grown in isolation &self pollinated to bring homozygosity in desirable traits. All important characters to be combined should be kept in mind.

79. 2. SELFING OF PARENTS ⚫2 nd step in hybridization .• Make the parents pure in its characters . Done by artificial self fertilization . Bisexual flowers covered using paper bags before anthesis to prevent natural cross pollination . Self pollination will occur inside the paper bags . In cross pollinated crops male & female flowers are bagged separately before anthesis .• When stigma becomes receptive, pollen grains are collected & brushed on stigma this is artificial pollination . Process continued till parent becomes homozygous or pure for its particular characteR

80. Conservation of medicinal plant

81. 3. EMASCULATION• 3 rd step in hybridization. Removal of stamens from female parent beforethey burst & shed pollen. Emasculation can be done by any of thefollowing methods FORECEPS AND SCISSORS METHOD Done in large flowers.Buds are opened & anthers removed using sterile forceps & needles or scissors No injury to be caused to other floral parts calyx, corolla & especially pistilb . HOT OR COLD WATER OR ALCOHOL TREATMENT For small flowers like that of rice, sorghum, etc. that are difficult to be emasculated by forn scissors, etc. Emasculation is done by dipping panicles in hot water for a definite period.Ideal temperature is between 450 C and 500C. Gynoecia can withstand a temperature at which anthers are killed.Also done by dipping inflorescence in cold water or alcohol for a definite period

82. MALE STERILITY In self pollinated crops, emasculation is eliminated by the use of male sterile plants in which anthers are sterile.Male sterility can be induced by spraying 2,4-D, NAA, maleic hydrazide , etc. on immature flower buds

83. 4. BAGGING After emasculation flower buds are kept enclosed in bags of ideal sizes, made of plastic, co or paper. Bags are tied by thread, wire, pins, etc.• This process is called bagging. Both male & female flowers bagged separately to prevent contamination (foreign pollen).Bagging is usually done before anthesis .

84. 5. CROSSING OR CROSS POLLIATNION⚫ 5 th step in hybridization. It is defined as artificial cross pollination between genetically unlike parents.• Viable pollen is collected from desired male plant & transferred on to the stigma of the desired emasculated female parent. Pollen grains are collected in petri dishes. • Bag is temporarily removed from the female parent & the collected pollen are dusted on stigma.

85. 6.LABELLING Emasculated & crossed flowers are properly bagged, tagged & labeled.Labeling is done either on bag itself or on separate labels. Labels are tagged to bags using thread. Label should bear following details – Reference number Date of emasculation Date of crossing Details of male and female parents

86. 7. COLLECTION OF HYBRID SEEDS After maturity of seeds, crossed heads of desirable characters are harv collected along with labels in separate envelopes. In the next season, seeds of each head are sown separately to raise FI generation.All plants of F1 are genetically similar & look exactly alike. They may exhibit hybrid vigor - increased growth, size, yield, function, etc. over the parents.

87. ➤ Significance of Hybridization The yield of crops can be increased by hybridization. By hybridizing different plants with better quality, a plant with superior quality can be produced Lacking disease resistance is the main problem in high-yield crops: this problem can be solved by producing disease-resistant plants by hybridization. By crosssing the plants with different tolerance, a variety with high tolerance to stres can be produced. Any desired characteristics like pest resistance, disease-resistant, and stress tolerance can be attained by hybridization. The senescence period can be extended and aging can be reduced in some high economic crops.