Enzymes in Biotechnology

It is:
- Industrial and commercial applications of biology - Usually involves enzymes, GE and micro-organisms

Anthony Wu

Enzymes as industrial catalysts:

- Advantages: Highly specific Efficient in small amounts Work at normal temperature & pressure

- Microorganisms > animals/plants to produce enzymes Higher growth rates & more enzyme per body mass More economical use low cost substrates Easily genetically engineered Some can grow at extreme temp & pH fully functional at these conditions

Enzyme immobilization:
- Advantages: Reusable Product is enzyme free (not contaminated) Cost-effective

- Disadvantages: Not as efficient as isolated

- Procedure: Prepare sodium alginate (2g) and distilled water (50cm3) mixture Prepare calcium chloride (1.4g) and distilled water (100cm3) mixture Mix together enzyme and sodium alginate solution Add drop by drop the mixture to the calcium chloride solution

Immobilised whole-cell enzymes (bacteria culture):

- Advantages: Good when enzymes are expensive Good when enzymes are difficult/tedious to obtain Stable - Disadvantages: Much substrate used by bacteria for growth rather than conversion

Optimum conditions to produce product may not be optimum conditions for bacteria growth

Immobilised cell-free enzymes (secreted by microorganisms):

- Advantages: Easy to obtain in bulk No wasteful side reactions - Disadvantages: Some may be tedious to extract

Industrial uses of enzymes:

Dairy Brewing Food - Cheese manufacture to coagulate milk proteins - Reduce win/beer cloudiness - Breakdown of starch to glucose for fermentation by yeast - Produce fructose syrup from glucose - Fruit juice increase volume of extracted juice, remove cloudiness from pectin - Pre-digestion of baby food - Meat tenderisation breakdown tough collagen & tissue - Rennin - Protease - Amylase - Glucose isomerase - Cellulase, hemicellulose - Pectinase - Trypsin - Bromelain - Papain - Ficin Textile Forestry & paper - Remove starch from fibres - Remove lignin from pulverised wood - Partial breakdown of starch smooth paper - Amylase - Ligninase - Amylase - Trypsin

Medicine - Remove blood clots in wounds

Properties of enzymes:
Work on specific substrates Remain unchanged at end of reaction reusable Biological catalysts Work best at optimum pH & temp Gets denatured at high temperatures

Biotechnology
Fuelling the future:
Biofuels

Anthony Wu

Feeding the world:

Natural pesticide More productivity o Prevent diseases o Drought resistant GM food

Revolutionising industry:
Cleaner, greener, more efficient Using enzymes o Plastic o Fermentation o Pasteurisation o Meat tenderisation, cleaning

Gathering information:
Genome sequencing o Human Genome Project (HGP) DNA computing

Making copies:
Biological materials Stem cells tissue Clone organisms (Dolly the sheep)

Environment care:
Biofuels renewable Bioindicators Clean oils pills

Diagnosing & treating illnesses:

Prosthetics Genetic screening o Customise treatment Vaccines from microorganisms

Chemicals of Life
Biological Molecules:
- Large complex molecules produced by living organisms

Anthony Wu

Carbohydrates:
- Made of elements: C, H, O - General formula: CX(H2O)X - 1g yields 16 kJ of energy - 3 types:

Monosaccharides (C6H12O6) o Glucose o Fructose o Galactose Disaccharides o Sucrose (cane sugar) o Lactose (milk sugar) o Maltose (malt sugar) Polysaccharides o Functions: Energy storage Ideally: Compact & inert Mobilised quickly when food is unavailable Starch & glycogen: Large & insoluble in water Foldable into compact shapes Easily converted to sugars by hydrolysis Structural support Cellulose plants Chitin fungi o Formed though enzyme-mediated dehydration synthesis o Also called glycans, differing in: Nature of recurring monosaccharides Length of their chains Degree of branching
- Dehydration synthesis/condensation reaction: Formation of complex molecule from the bonding of 2 simpler molecules with the removal of a molecule of water

E.g.: Glucose + Fructose Sucrose + Water Glycosidic bond between monomers

- Hydrolysis: Opposite of dehydration synthesis: o Sucrose + Water Glucose + Fructose

- Function: Source of energy Form supporting structures (cellulose, chitin) Formation of nucleic acids (DNA & RNA) Synthesize lubricants (mucus, etc.) Produce nectar in flowers to attract insects for pollination

- Benedicts test: For reducing sugars (All monosaccharides and disaccharides except sucrose) Add equal volume of benedicts solution and sample. Shake. Heat solution in boiling water bath Positive test: coloured precipitate observed: o Green (less) Yellow Orange Red Brick-red (more) Negative test: Solution remains blue

- Iodine test: For starch Add a few drops of iodine solution to unknown sample Positive test: Blue-black mixture observed Negative test: Mixture remains yellowish-brown

Lipids:
- Made of elements: C, H, O - No fixed molecular formula/ratio - Much more H than O - 1g yields 38 kJ of energy - Types: Animal & vegetable fats o Triglyceride: 3 fatty acids + 1 glycerol molecule Formation dehydration synthesis

Breakdown Hydrolysis Forms ester bonds between glycerol & fatty acids

Phospholipids o Constituent of cell membranes Steroids (e.g.: cholesterol) Saturated fats: o No double bonds present in fatty acid chain o Found in animals (pork, beef etc.) Monounsaturated fats: o Presence of 1 double bond in fatty acid chain o Examples (Olive oil, peanut oil, avocado, salmon, mackerel, trout etc.) Polyunsaturated fats: o Presence of 2 or more double bonds in fatty acid chain o Found in many nuts (Sunflower oil, corn oil, soybean oil, almonds, cashews, walnuts, macadamia)

- Function: Source & store of energy Insulating material prevent heat loss Solvent for fat-soluble vitamins (Vitamin A, D, E, K) & hormones (sex hormones etc.) Constituent of cell membranes (phospholipids) Restrict water loss from skin surface Production of sex and growth hormones from cholesterol

- Overconsumption: Increase in blood levels of cholesterol Excess cholesterol deposits on inner walls of arteries atherosclerosis High blood pressure & blood clot formation blockage in coronary arteries heart attack

- Ethanol-emulsion test: For all lipids Add 2cm3 of ethanol to a drop of sample oil. Shake thoroughly Add 2cm3 of water to mixture and shake. Positive test: White milky emulsion formed & heat is evolved Negative test: No emulsion observed

Proteins:
- Made of elements: C, H, O, N, (sometimes S & P) - Basic unit = amino acid - Dehydration synthesis forms larger structures dipeptides, oligopeptides, polypeptides etc. - 1g yields 17 kJ of energy - Structure: Amino group + R group + Carboxyl group R groups: Group Non-polar Polar Acidic Basic - Sources: Animal o Meat, eggs, milk, seafood, liver Plant o Peas, beans, nuts Characteristics Hydrophobic Hydrophilic Negatively charged Positively charged Names Ala, Val, Leu, Ile, Pro, Phe, Trp, Met Gly, Ser, Thr, Cys, Tyr, Asn, Gln Asp, Glu Lys, Arg, His Example Leu: CH2CH(CH3)2 Thr: CH(OH)CH3 Asp: CH2COOH Lys: (CH2)4NH3+

Linkages peptide bonds

- Function: - Types: Globular proteins o Transport protein haemoglobin (transport of oxygen from lungs to body), membrane pumps (transport molecules across cell membranes) o Enzymes speed up rate of chemical reactions o Antibodies destroy invading bacteria (immunity) Structural proteins Synthesis of protoplasm (i.e.: nucleus, cytoplasm, cell membrane) repair and growth of body cells Synthesis of enzymes and some hormones (e.g.: insulin) Formation of antibodies to combat diseases Source of energy

o Collagen (component of bone, tendon, teeth, skin) o Keratin (hair & nails) - Biuret test: Add 1cm3 of sodium hydroxide solution to 2cm3 of protein solution. Shake thoroughly Add copper sulphate solution to mixture, drop by drop, shaking after each drop Positive test: Violet/purple colouration is observed Negative test: solution remains blue

Summary of molecules:
Class Carbohydrate Lipids Proteins Nuclei Acids Monomer Monosaccharides (e.g.: Glucose etc.) Fatty acids; Glycerol Amino acids Nucleotides Polymer Polysaccharide (e.g.: starch etc.) NA Protein (e.g.: Haemoglobin etc.) DNA, RNA

Water:
- 70-80% of cells made of water - Universal solvent most common medium of chemical reactions (e.g.: hydrolytic reactions of digestion) - Transporting agent for digested food substances, hormones & excretory products from one part of the body to another - Essential component of lubricant (e.g.: in joints, blood & digestive juices) - Raw material for photosynthesis - Temperature-regulation excess body heat removed through evaporation

Photosynthesis & Leaf Structure

Structure of typical leaf:
- 3 Parts: Leaf blade (Lamina) Leaf stalk (Petiole) Leaf veins - 2 types: Simple leaf Compound leaf

Anthony Wu

- Characteristics: Lamina: o Large flat surface Increase surface area & maximise exposure to sunlight o Thin Allow carbon dioxide to reach inner cells rapidly Enables sunlight to reach all mesophyll cells Petiole: o Hold leaf away from stem Absorb maximum sunlight o Continues into leaf blade o Sessile leaves no petiole Veins: o o o o Carry water and mineral salts to cells in lamina Carry manufactured food from leaf blade to others parts of the plant Dicotyledonous plants network veins, lateral roots Monocotyledonous Parallel veins, fibrous roots

- Internal structure:

Epidermis: o Thin, waxy cuticle layer reduces water loss o Single layer of closely packed cells o Keeps leafs shape o Reduces evaporation from the leaf, preventing bacteria and fungi from getting in o Focuses light on mesophyll layers Mesophyll: o Below epidermis o Site of photosynthesis o Split into 2 regions: Palisade tissue Long, contain many chloroplasts Chloroplasts absorbs suns energy to make carbohydrates from carbon dioxide and water Spongy tissue Irregularly shaped Loosely arranged to created air spaces among them allow rapid diffusion of carbon dioxide Also contains chloroplasts

o Chloroplasts: Oval shaped containing chlorophyll Grana: Stacks of membranes or thylakoids Light dependent reactions sited here Stroma: Enzymes for light independent stage here Stomata: o Found in epidermis (more on lower in most dicots) o Consists of a pair of guard cells surrounding a stomatal pore o Guard cells bean-shaped, contain chloroplasts o Regulation: During sunlight hours, potassium ions enter guard cells decrease water potential Water enters guard cells via osmosis Guard cells swell and become turgid (one side has thicker cellulose cell wall) pore opens

At night, potassium ions leave guard cells water potential

increases Water leaves guard cells Guard cells become flaccid pore closes o Regulation controls water loss o Entry of carbon dioxide into leaf: CO2 used up during photosynthesis concentration decreases Diffusion gradient exists CO2 diffuses through stomata into system of air space in leaf CO2 dissolved in thin film of water on mesophyll layer cells diffuse into solution of cells o Water from xylem leaf Water and dissolved mineral salts diffuse from cell to cell of mesophyll cells after exiting veins

Photosynthesis:
- Definition: Light energy absorbed by chlorophyll and transformed into chemical energy used in the synthesis of carbohydrates from water and carbon dioxide. Oxygen Is liberated in the process exact phrasing

- Importance: Converts light energy from sun to chemical energy stored in carbohydrates Carbohydrates, proteins, fats form other organic compounds fills food requirements Energy in coal, petroleum & natural gas comes from sun via photosynthesis Purifies air by removing carbon dioxide and releasing oxygen

- Heterotrophs: Unable to make own food feed on other organisms Completely dependant on photoautotrophs for food & oxygen

- Conditions: Chlorophyll Light Carbon dioxide Water Temperature (enzymes light independent)

- Light dependent stage: Energy from sun absorbed by chlorophyll and converted to chemical energy Light energy splits water molecules to oxygen & hydrogen 12H2O (photolysis) 6O2 + 24H - Light independent stage: Hydrogen used to reduce carbon dioxide to carbohydrates (glucose) Light dependent stage provide energy 6CO2 (energy) C6H12O6 + H2O - Equations: Carbon dioxide + Water (chlorophyll, light) Glucose + Oxygen 6CO2 + 12H2O (chlorophyll, light) C6H12O6 + 6H2O + 6O2

- Limiting factors: Any factor that directly affects a process if its quantity is changed Photosynthesis: o Concentration of CO2 (most important under normal environmental conditions) o Light intensity o Temperature Used in tissue respiration to provide energy for cellular activities Make cellulose cell walls Excess converted to sucrose, transported to storage organs as starch or other forms In large amounts, temporarily stored as starch in leaves In darkness, photosynthesis stops and starch is reconverted by enzymes to simple sugars Fats formed from glucose Reacts with nitrates & other mineral salts to form amino acids, which combine to form proteins

- Glucose:

Excess amino acids stored as proteins build new protoplasm in leaves or growing plant parts - Enzymes used: Starch Maltose diastase Maltose Glucose maltase Proteins Peptones pepsin Peptones Amino acids erepsin Fats Fatty acids and glycerol lipase

Respiration Cellular respiration

Definition:

Anthony Wu

- Oxidation of food substances releases energy - Occurs in living cells energy released in the form ATP (Adenosine Triphosphate) - 3 stages: Glycolysis o Occurs at cytosol of cell o Input: Glucose + 2ATP o Output: 4ATP + 2NADH + 2H2O + 2 pyruvate Krebs Cycle/tricarboxylic cycle/citric acid cycle o Occurs at mitochondrial matrix o Input: Pyruvate o Output: 2ATP + 8NADH + 2FADH2 + 6CO2 Oxidative phosphorylation o Occurs on inner membrane of mitochondria o Input: 10NADH + 2FADH2 o Output: 34ATP + H2O

Energy:
- Uses Synthesis: o Formation of new substances for growth, development & repair Transport: o Active transport o Movement of materials across cell membranes Movement: o Muscular contractions Electrochemical activity: o Generation of nerve impulses Heat production: o Maintain constant body temperature (warm-blooded animals)

Adenosine Triphosphate (ATP):

- Universal energy currency - Constantly recycled - Contains a lot of chemical energy - Takes part in many metabolic reactions

- Delivers energy in small amounts to drive individual reactions - Involved in exergonic and endergonic reactions - Formation: Respiration & photosynthesis - ATP-ADP cycle: ATP ADP & P + energy to do work o Active transport o Building molecules o Muscle contraction o Working of neurons

Aerobic respiration:
- Respiration in presence of oxygen - Releases lots of energy - Takes place in mitochondria of cells - Equation: Glucose + oxygen Carbon dioxide + water + lots of energy C6H12O6 + 6O2 6CO2 + 6H2O + lots of energy

Anaerobic respiration:
- Respiration in absence of oxygen - Releases a small amount of energy - Yeast cells anaerobic respiration: Ethanol & carbon dioxide produced fermentation o Used in wine making, bread making Glucose Carbon dioxide + ethanol + small amount of energy C6H12O6 2CO2 + 2C2H5OH + small amount of energy

- Anaerobic respiration less efficient than aerobic: Small amount of energy produced Lactic acid & ethanol produced o Harmful to organisms if accumulated o Contains much unused energy Lactic acid can be converted back to sugar to be used for respiration Yeast cannot metabolise ethanol

- Anaerobic respiration in muscles: Muscular activity lack of oxygen in muscles Respiration not complete Lactic acid (CH3CHOHCOOH) built up Only first stage of respiration occurs (i.e. Glycolysis) 2ATP produced Accumulation of lactic acid muscle fatigue After rest [Cori cycle]: o Lactic acid transported to liver o Some oxidised to produce energy convert remaining lactic acid to glucose o Glucose transported to muscles for use Oxygen debt occurs o Amount of oxygen required to oxidise the lactic acid produced in muscles during anaerobic respiration Glucose Lactic acid + small amount of energy C6H12O6 2CH3CHOHCOOH + 4H + small amount of energy

Respiration Energy liberated O2 used, CO2 & H2O released Catabolic process, breakdown of glucose Occurs all the time Results loss of dry mass

Photosynthesis Energy stored in carbohydrate molecules CO2 & H2O used, O2 given off Anabolic process ; glucose is formed Occurs in presence of sunlight and chlorophyll Results gain of dry mass

External respiration
Inspiration:
- Process in which air is taken into the body - Principle: Thoracic cavity expands Lungs expand to fill up enlarged space Expansion = lungs air pressure < atmospheric pressure Air rushes into lungs

- Breathing movements: External intercostal muscles contract internal intercostal muscles relax Ribs swing upwards & outwards Sternum moves up and away from vertebral column

^ Increases dorso-ventral diameter & breadth of thorax Diaphragm contracts & flattens ^ Enlarges thoracic cavity Volume of thorax increases air pressure decreases air enters lungs

Expiration:
- Process in which air is given out from the body - Principle: Thoracic cavity contracts Lungs contract Contraction = lungs air pressure > atmospheric pressure Air rushes out of lungs

- Breathing movements: External intercostal muscles relax internal intercostal muscles contract Ribs lowered Sternum moves back near vertebral column Diaphragm relaxes & arches upwards ^ Decreases thoracic cavity Volume of thorax decreases air pressure increases air forced out of lungs

Lungs:
- Organ dedicated to the exchange of gases between man and the environment - 2 lungs present - Located in the chest cavity - Path of air to lungs: Nasal passages o Air enters through nose/mouth o Breathing through nose Dust & foreign particles trapped by hair in nostril & mucus on mucous membrane Air warmed and moistened Harmful chemicals detected by sensory cells in mucous membrane

Pharynx o Air passes into pharynx from nasal passage Larynx [voice box] o Extends to chest cavity o Supported by C-shaped rings of cartilage

Trachea o Air passes to trachea through an opening, the glottis o Divides into left and right bronchus o Supported by C-shaped rings of cartilage o Epithelium lining bears gland cells Secretes mucus trap dust particles & bacteria Bear cilia sweep dust particles into pharynx Bronchi o Left bronchus divides into 2 bronchial tubes o Right bronchus divides into 3 bronchial tubes o Supported by C-shaped rings of cartilage o Epithelium lining bears gland cells Secretes mucus trap dust particles & bacteria Bear cilia sweep dust particles into pharynx

Bronchioles o Ends in cluster of alveoli Alveoli o Thin, moist and well vascularised walls o Site of gaseous exchange o Indicates large surface area for gaseous exchange

- Other structures: Ribs o o o o Supports chest wall Front attached to sternum (chest bone) Back attached to vertebral column (backbone) 12 pairs of ribs, first 10 attached to sternum

Intercostal muscles o External and internal muscles between ribs o One set contracts, other set relaxes o Moving up and down change volume of thoracic cavity

- Gaseous exchange: Air entering lungs more oxygen, less carbon dioxide Blood entering lungs less oxygen, more carbon dioxide ^ Diffusion gradient present, maintained by o Continuous flow of blood through blood capillaries o Continuous flow of air through alveoli

Membrane separating blood capillaries from alveolar air permeable to both gases Oxygen: o Dissolves in moisture lining alveolar walls Why? To prevent formation of air bubble could cause stroke o Dissolved oxygen diffuses into blood o Combines with haemoglobin to form oxyhaemoglobin Haemoglobin contains porphyrin ring structure (heme group) with ferrous iron (Fe2+) covalently binded, which reversibly binds O2 Hb + 4O2 HbO8 Carbon dioxide: o H2O + CO2 H2CO3 H+ + HCO3o When CO2 concentration is low, carbonic anhydrase catalyse reaction in which hydrogen carbonates converted to carbon dioxide and water o % transported: Dissolved in plasma (7%) Bound to haemoglobin (23%) Dissolved as bicarbonate in plasma (70%) o Diffuses out of blood into alveolar cavities o Water evaporates from walls of alveoli Expired air 16% oxygen 4% carbon dioxide 78% nitrogen Water vapour saturated Body temperature (~37o) Dust particles absent

Inspired air 21% oxygen 0.03% carbon dioxide 78% nitrogen Water vapour variable Temperature variable Dust particles may be present

Gills:

- Countercurrent mechanism: Water flows in opposite direction to blood Concentration gradient of oxygen is maintained Diffusion is more efficient and fast

Plants Transport in plants

Xylem:
- Vascular tissue that transports water & mineral salts - From roots to leaves of the plants - Provides mechanical support to the plants - Xylem cells dead at functional maturity

Anthony Wu

No protoplasm, hollow provide unimpeded flow of water No complete end walls water can flow easily from one cell to the next - Lignin deposited on cell wall Hard & rigid to provide support & prevent collapse

Phloem:
- Vascular tissue that transports sugars and other food substances - From photosynthesizing/storage regions to other plant parts - Photosynthesizing & storage regions high amount of sugars Photosynthesizing parts glucose Transport sugar sucrose Storage sugar starch - Made of sieve tubes cells stacked upon one another Degenerate protoplasm (lost nucleus & most organelle & almost all central vacuole) provide unimpeded flow of sugars

Sieve plates with sieve pores between cells to allow flow of substances - Companion cell associated with each sieve tube cell Dense cytoplasm (lots of mitochondria) - Provide energy for sieve tube to survive & carry out active transport

Monocotyledons vs. Dicotyledons:

- Based on no. of cotyledons found in seeds Seed leaves Monocots (e.g.: maize, orchids, rice, garlic, onion) Dicots (e.g.: rose, hibiscus, magnolia, apple, avocado) One cotyledon Two cotyledons Veins in leaves Parallel Netlike Vascular bundles Scattered Arranged in ring Flower parts Multiples of threes Multiples of fours and fives

Stems:
- Epidermis - Cortex - Pith Stores food & provides plant support Made of parenchyma cells Outer layer (waterproof) Stores food & provides plant support Made of parenchyma cells

- Phloem + Xylem = Vascular bundle Monocots Arrangement of vascular bundle Cambium (produces xylem & phloem cells) Pith Scattered about in cortex Absent Absent Dicots Arranged in a ring Present Present

Roots:
- Casparian strip Controls entry of substances into xylem Waterproof - Pericycle Gives rise to branch roots - Endodermis Delineates a boundary for the stele - Casparian strip + Pericycle + Endodermis = Stele Monocots Arrangement of tissue Xylem & Phloem arranged alternate with each other in a ring Central Pith Present Dicots Xylem found in the middle in a 4-5 pronged star shape with phloem arranged alternatingly Absent

Leaves:
- refer to leaves notes Monocots Mesophyll layer Stomata No differences Both sides Dicots 2 distinct regions palisade & spongy Only underside

Path of water & minerals:

- Water around soil particles root hairs xylem of roots xylem of leaves - Proof xylem transports water: If leaf shoot kept in dyed water is cut, only xylem is stained Plant only wilts when xylem is removed, not phloem Mineral ions found in xylem sap

- Water from soil Root hairs: Water enters plant through epidermal extensions (root hairs) at piliferous layer Layer of dilute salt solution around root hair cells Water enters root hair cell by osmosis due to presence of potential gradient Ions enter root hair cell via diffusion & active transport lower water potential Root hair cells increase surface area to volume ratio for efficient absorption Root hair cell contains a lot of mitochondria provide ATP for active transport

- Root hairs Xylem: Water must move through cells to reach xylem Moves cell to cell through cortex, endodermis & Pericycle by osmosis to reach xylem Mineral ions move via diffusion 3 paths: o Apoplastic (continuum of cell walls) o Symplastic (continuum of cytosol through plasmodesmata) o Transmembrane (from vacuole to vacuole through cell wall) Casparian strip (waterproof) o Water & dissolved ions cannot pass through apoplastically, only symplastically or transmembrane o Acts as selective sentry, controlling transport of certain materials

- Xylem Leaves (3 ways): Root pressure: o Root cells pump ions into xylem actively decreases xylem water potential o Water enters by osmosis, pushes xylem sap upwards o Only useful for small plants, ineffective in large trees Capillary action: o In narrow vessels, water moves up by capillary action because of Adhesive forces between water molecules & walls of vessel

Cohesive forces between water molecules

o Why? Water has high surface tension strong hydrogen bonds o Only up to 3m, ineffective in large trees Transpiration pull: o Loss of water vapour from aerial parts of plant (esp. leaves) o Main factor that transports water to leaves o Steps: Water leaves mesophyll cells to form a layer of moisture around the cells Moisture evaporates moves into intercellular air spaces Water vapour escapes by diffusion via stomata Evaporation causes mesophyll to become try Mesophyll cells absorb water from neighbouring cells absorb from xylem Results in suction force pulling column of water up xylem vessels (cohesive & adhesive forces) Effective for over 100m

Path of food substances:

- Process is active requires ATP - Proof phloem transports food: Carbohydrates exudes from cut phloem Removal of phloem accumulation of sugars Radioactive tracing using 14CO2 Material in feeding aphids stylets are sugars

- Translocation: Transport of manufactured food substances Bidirectional Much controversy on mechanism for translocation

- Mass Flow Hypothesis Source = Photosynthetic organ capable of producing sugar Sink = Organ that stores/consumes sugar Loading sugars into source decreased water potential water enters by osmosis Created high pressure at sources compared to sink sap flow towards sink Sugars in sink increased water potential water leaves to xylem by osmosis

Summary: Sugars move from source to sink, water recycled from xylem Experimental evidence: o Aphids along phloem severed at stylet o Closer to source sugar exuded out faster Limitations: o Does not explain existence of sieve plate impedes flow of sugars

Transpiration:
- Importance: Draw water & minerals up from roots stem leaves o Water used in photosynthesis o Water keeps plant cells turgid keeps leaves spread out to trap sunlight

Evaporation cools plant loss of latent heat of vaporisation - Measuring: Use potometer o Measures distance air bubble moves over time - Excessive transpiration Transpiration rate > absorption rate mesophyll cells lose turgor wilting Wilting decreases exposed surface area to sun o Advantages: Flaccid guard cells Closed stomata reduce transpiration

rate o Disadvantages: Insufficient H2O & lack of CO2 reduced photosynthesis rate - Factors affecting transpiration: Cuticle: o Thick Waterproof cuticle makes it harder for water to pass through Transpiration rate decreases o Thin Thinner layer makes it easier for water to pass through Transpiration rate increases Stomata: o Closed Closed stomata prevents water vapour from escaping Transpiration rate decreases o Open Larger opening enable water to transpire easily Transpiration rate increases

Boundary layer of air: o Thick Slow diffusion of water vapour lower concentration gradient Transpiration rate decreases o Thin Water vapour diffuses faster higher concentration gradient Transpiration rate increases Humidity: o High o Low

High humidity decreased concentration gradient Transpiration rate decreases Low humidity increased concentration gradient Transpiration rate increases

Temperature: o Low Low temperatures air holds less water Reduces concentration gradient Transpiration rate decreases o High High temperatures air holds more water Increases concentration gradient Transpiration rate increases

Light: o Dark o Bright

Stomata close in dark lack of photosynthesis Transpiration rate decreases Stomata open in light photosynthesis Transpiration rate increases

Wind: o Weak Weak/no wind cannot move boundary layer created by water vapour Transpiration rate decreases o Strong Strong wind moves boundary layer created by water vapour Transpiration rate increases

Plant adaptations:
- Xerophytes: Adapted to survive in environments with little water Reduce rate of transpiration o Thick cuticle forms waxy barrier preventing water loss E.g.: Most cacti o Absence of true leaves limits water loss to stems only (fewer stomata) E.g.: Most cacti o Reduction of surface area to volume ratio of leaves limits transpiration area E.g.: Pine o Sunken stomata moist air trapped there lengthens diffusion pathway reduces evaporation & transpiration rate E.g.: Sorghum Storage of water o Succulent leaves & stems stores water E.g.: Bryophyllum, most cacti o Extensive root system allows for efficient absorption of water over wide area - Hydrophytes: Adapted to survive wholly/partially submerged in water Obtain oxygen & water o Location of stomata floating plants only have on upper surface of leaf, absent in submerged plants oxygen directly absorbed through diffusion E.g.: Water lilies o Cuticle absent in submerged plants, only at floating plants top surface E.g.: Hydrilla thin cuticle, Water lilies top surface only (thick) o Presence of xylem redundant, water can easily diffuse directly, flexible stems might snap in strong current E.g.: Absent in Hydrilla o Presence of aeration (aerenchyma) Large air spaces between leaf cells for buoyancy for plants to get more sun E.g.: All floating plants o Root system Reduced root system, used for anchorage only E.g.: Most hydrophytes E.g.: Most succulent plants