core practicals 1-13

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what is plasmolysis?

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1

what is plasmolysis?

when the protoplasm of a plant cell begins to shrink away from the cell wall because the water potential and the osmotic potential are equal

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2

outline the procedure to determine the water potential of a plant tissue

  1. transfer a small set volume of each mineral salt solution into a watch glass and leave for 20 mins

  2. remove each section using forceps. mount in a drop of the corresponding solution on a microscope slide and cover with a coverslip

  3. observe 25 cells and record how many are plasmolysed

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3

when can a solution be determined to be isotonic?

when the line of best fit crosses the x axis

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4

what does isotonic mean?

when the water potential of the mineral salt solution is the same as the water potential of the plant tissue, so there is no net movement of water

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5

what is incipient plasmolysis?

where the water potential of the cytoplasm is the same as the solution - the point at which plasmolysis just begins to occur - when 50% of the cells in a tissue are plasmolysed

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6

why are the plant cells left in the mineral salt solution for 20 minutes before observation?

to allow time for osmosis to reach equilibrium

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7

why is aseptic technique important?

creates reliable and repeatable data avoids contamination of the sample from outside substances

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8

what is streak plating?

bacteria are spread out on a nutrient agar plate so that distinct individual colonies can be seen colonies grown on clean agar plates to produce non-contaminated samples of one species of bacteria

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9

how can colonies be identified as a particular species of bacteria?

size, colour, and texture of the colony

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10

disinfectant risk assessment

hazard - disinfectant risk - flammable safety precaution - keep away from naked flame in emergency - put out fire, seek assistance

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11

what parts of aseptic technique are used in cp13?

flame sterilise the inoculating loop before and after use flame sterilise the neck of the bottle the mixed culture is in after opening and before closing open the petri dish lid as little as possible

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12

why should the petri dish lid not be taped the whole way around?

so the conditions are not anoxic and the bacteria on the agar can actually grow

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13

how can the colonies produced in cp13 be identified?

white colonies are likely to be salmonella yellow colonies are likely to be staphylococcus

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14

outline the procedure to isolating a particular species of bacteria

  1. dip a sterile inoculating loop into the mixed culture

  2. make streaks on the agar using the inoculating loop

  3. incubate the agar plate for 24 hours

  4. take a sample of the different colonies and streak them on different agar plates and incubate again for 24 hours

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15

what is aseptic technique?

procedure used to avoid contamination of the sample from outside substances, such as microorganisms

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16

how is aseptic technique used in cp12?

wipe down surfaces with antibacterial before & after use bunsen burner to create sterile zone around workspace flame sterilise wire loop and necks of any bottles keep vessels containing bacteria open for minimum amount of time close all windows and doors

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17

what is chromatography used for?

separate and identify small quantities of different compounds

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18

what are the factors affecting the rate of migration?

solubility mass affinity to the paper

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19

outline the procedure for cp11

  1. draw pencil line 1cm above bottom of paper

  2. cut section of leaf and grind in pestle and mortar with 20 drops of propanone to release pigments

  3. use capillary tube to extract pigment and blot on paper

  4. leave paper in solvent for 10 minutes

  5. take paper out of solvent and draw a pencil line where pigments moved to

  6. calculate Rf value for each spot of pigment

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20

chromatography solvent risk assessment

hazard - chromatography solvent risk - flammable, irritant, harmful to inhale safety precaution - avoid contact with solvent, wear eye protection, keep away from flame in well-ventilated room in emergency - wash skin/eyes using cold water, put out flame, seek medical attention

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21

acetone risk assessment

hazard - acetone risk - flammable, irritant, inhalation can cause dizziness and drowsiness safety precaution - avoid contact, wear eye protection, keep away from naked flame in well ventilated room in emergency - wash skin/eyes with water, seek medical assistance

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22

how can pigments be identified?

comparing the Rf values calculated to known Rf values using same chromatography paper and solvent

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23

why does affinity affect the rate of mobility?

pigments have different affinities to chromatography paper, those with lower affinities will travel further up the paper

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24

why does solubility affect the rate of mobility?

pigments that are more soluble travel further up the paper and end up closer to the top at the solvent front

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25

Rf value formula

distance moved by pigment / distance moved by solvent

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26

outline the procedure for cp10

  1. place a piece of pondweed in a beaker of water

  2. cover one side of the beaker with aluminium foil to block out light

  3. cover the other side of the beaker with a coloured light filter

  4. add half a spatula of sodium hydrogencarbonate to the water to provide carbon dioxide

  5. leave for 5 minutes

  6. place the bench lamp a set distance from the beaker e.g. 30cm

  7. measure the movement of the air bubble up the capillary tube

  8. replace the coloured light filter with another and repeat the experiment

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27

sodium hydrogencarbonate risk assessment

hazard - sodium hydrogencarbonate risk - contact with eyes, inhalation safety precaution - wear goggles, avoid contact with skin in emergency - seek medical attention, flood cuts/eyes with cold water

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28

why does a green filter decrease the volume of gas the most?

chloroplasts don't absorb much green light - they reflect it, giving them their green appearance

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29

when is the greatest volume of gas produced?

when there is no filter, because all wavelengths of light can be absorbed

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30

what does the volume of gas indicate?

assumed to be oxygen - volume of gas produced is directly proportional to the rate of photosynthesis

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31

cp10 control variables

light intensity amount of sodium hydrogencarbonate time allowed for gas collection temperature

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32

what is a respirometer?

a piece of equipment used to measure the rate of respiration

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33

how does a respirometer work?

adding a drop of coloured liquid to the length of tubing. as the organism respires and the volume of oxygen in the tube decreases, the pressure also decreases and the liquid moves down the tube towards the respirometer

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34

outline the procedure for cp9

  1. assemble the respirometer

  2. add 5g of one organism to the boiling tube and replace the bung

  3. place a drop of coloured liquid in the open end of the respirometer and use a syringe to draw the liquid as far back as possible from the respirometer and record the starting position

  4. close the tap and start the stop clock

  5. open the tap after 5 minutes and record the position of the tube

  6. repeat the process with another organism or a control

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35

soda lime risk assessment

hazard - soda lime risk - corrosive safety precaution - wear eye protection, avoid contact with skin in emergency - wash off skin immediately, flood eyes/cuts with cold water

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36

what factors can affect the results of cp9?

change in temperature or atmospheric pressure handling the apparatus may add heat and change the readings

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37

what is used to absorb carbon dioxide in cp9?

soda lime

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38

how do you find the volume of oxygen consumed using a respirometer?

measure the distance moved by the coloured liquid, and use the formula volume = distance x pi x r2

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39

how is the rate of respiration measured using data from a respirometer?

volume of oxygen used / mass / time

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40

cp9 control variables

mass of organism time temperature mass of soda lime apparatus must be air-tight - replace air between each set up

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41

what are the ethical issues surrounding this practical?

the way the locusts are raised and killed is killing a living thing wrong? is it justified in the name of science?

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42

disinfectant risk assessment

hazard - disinfectant risk - flammable safety precaution - keep away from open flames in emergency - put out fire, seek assistance

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43

outline the procedure for dissection in cp6

  1. pin the locus to the dissection board and remove the exoskeleton

  2. identify the internal gas exchange system. flood the specimen with water so the tracheae show silvery-grey

  3. examine the tracheae under a microscope

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44

what is a potometer?

a device used to measure the uptake of water, and therefore the rate of transpiration, in plants

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45

what affects the rate of transpiration?

light intensity, humidity, wind speed, and temperature

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46

lamps risk assessment

hazard - lamp risk - temporary damage to eyes safety precaution - do not look directly at lamp in emergency - wait for afterimage to disappear, seek appropriate assistance if needed

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47

bags risk assessment

hazard - bag risk -tripping safety precaution - keep under desks and away from workspace in emergency - seek appropriate medical assistance, clean spillages

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48

why does light intensity affect rate of transpiration?

affects the rate of photosynthesis, which affects the amount of stomata open

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49

why does temperature affect the rate of transpiration?

increases the rate of diffusion and evaporation from the stomata

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50

why does humidity affect the rate of transpiration?

affects the rate of diffusion and evaporation by decreasing concentration gradient between plant and the atmosphere

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51

why does wind speed affect the rate of reaction?

increases concentration gradient by mechanically removing water from the outside the stomata

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52

betalain

red, water-soluble pigment in beetroots found in vacuole of each cell

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53

how can the permeability of a membrane be changed?

temperature concentration of solvents e.g. ethanol

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54

how can permeability of a beetroot cell membrane be measured?

amount of pigment leaked through the cell membrane into an aqueous solution using a colorimeter

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55

ethanol risk assessment

hazard: ethanol risk: irritant/flammable safety precaution: wear eye protection, keep away from naked flames in emergency: wash eyes and skin with cold water

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56

what is the effect on cell membrane permeability as the temperature increases and why?

permeability increases because proteins in the membrane denature and heat damages the bonds making their tertiary structure. this creates gaps in the membrane, making it easier for molecules to pass through it

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57

why do low temperatures cause a decrease in membrane permeability?

phospholipids have little energy and are packed closely together to make the membrane rigid - decreases permeability and restricts molecules from crossing the membrane

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58

outline the procedure to investigate the effect of temperature on permeability of cell membrane?

  1. cut beetroot into 6 identical cubes with a scalpel

  2. place each cube in a different test tube with equal volumes of distilled water

  3. place each test tube into water baths ranging from 30oC-80oC. leave for 20 minutes

  4. filter each solution out into a cuvette and measure the absorbance

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59

what is the effect of ethanol concentration on membrane permeability?

increasing ethanol concentration leads to increased membrane permeability, as ethanol causes gaps to form in the membrane

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60

biohazard risk assessment

hazard: biohazard risk: allergies, soil bacteria, contamination safety precaution: wash hands after practical in emergency: seek assistance

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61

what happens as sucrose concentration increases?

mean pollen tube growth also increases up to an optimum. after this point, as sucrose concentration increases, mean pollen tube growth decreases - osmotic effects of increasing concentrations of sucrose

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62

what are the control variables of this practical?

environment - humid chamber volume and content of nutrient salt solution volume of sucrose solution time allowed for growth

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63

outline the procedure to cp4

  1. dilute the stock sucrose solution to several set concentrations

  2. place a moist piece of filter paper into a petri dish to form a humid chamber

  3. put a few drops of sucrose solution and an equal volume of mineral salt medium onto a clean microscope slide

  4. use a mounted needle to rub the anther of the flowers so they shed some pollen onto the microscope slide

  5. place the slides into the petri dish until it is time to observe them

  6. start the stop clock. place the slides under the microscope and use a calibrated eyepiece graticule to measure pollen tube growth

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64

why should a cover slip not be used?

to allow oxygen to reach the pollen and prevent conditions from being anoxic

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65

what is the role of gibberellins during germination?

it is secreted by the embryo, then it diffuses to the aleurone layer of the endosperm to stimulate the production of amylase. amylase hydrolyses starch to maltose

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66

how is the effect of gibberellin concentration on amylase production investigated?

  1. dilute gibberellin to produce several concentrations

  2. cut seeds in half, use only the half with the endosperm

  3. dip in sodium hypochlorite solution and wash with water

  4. place seeds in each gibberellin solution and leave

  5. place seeds in a petri dish and starch agar, and leave for 12-48 hours

  6. pour potassium iodide onto the plates and measure the clear zone

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67

what does the zone of inhibition indicate?

starch has been hydrolysed by amylase

the larger the zone of inhibition, the higher the amylase concentration

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68

why are the seeds placed in sodium hypochlorite solution?

to sterilise the seeds

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69

what is the effect of gibberellin concentration on amylase production?

increasing gibberellin concentration increases the area of the clear zone, indicating an increased production of amylase and more starch hydrolysed

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70

what are some sources of error in cp14?

the existing gibberellin and amylase content of the seeds may be different

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71

control variables of cp14

  • time allowed to soak in gibberellin

  • time left on starch agar plate

  • source/age of seeds

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72

what is the independent variable of cp15?

sampling method - using a frame, point quadrat, quadrat size

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73

how is percentage cover calculated?

  • use a quadrat with squares

  • count how many squares the required species is present in

  • divide this by the total number of squares available

  • multiply by 100 to convert into a percentage

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74

outline the procedure to cp15

  1. choose an area to take samples from. use two tape measures to create a set of axes off which coordinates can be read

  2. use a random number generator to generate 10 sets of random coordinates

  3. place the quadrat at each of the coordinates, placing the bottom left corner on the coordinate every time. start with the smaller quadrat

  4. record the percentage cover for the chosen species

  5. record the abundance of each species within the quadrat. this is used to work out density

  6. record how many pins touch the chosen species. if it’s a frame quadrat work out how many quadrat intersection points contain the chosen species underneath.

  7. repeat steps 3-6 with the bigger quadrat

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75

outline the procedure to cp16

  1. choose an abiotic factor

  2. choose an area that has a clear light intensity gradient

  3. lay a 20m tape measure on the ground from the area with sunlight to the shaded area

  4. choose a species that changes in abundance along your transect

  5. place a quadrat at the 0m mark

  6. measure light intensity at the 0m mark within the quadrat

  7. record the abundance of your chosen species by counting how many organisms are present. record this in a table

  8. repeat steps 5 and 6 every 2m along the tape measure until you reach the end of the 20m

  9. repeat steps 3-7 by creating another 2 transects between the area with sunlight to the shaded area

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76

how else can you measure abundance?

working out percentage cover

estimate - less accurate

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77

what is the difference between a risk and a hazard?

the hazard describes the potential source of harm, whereas the risk describes the likelihood of the harm occurring

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78

how can the results be used to determine the relationship between the chosen factor and the morphology of a species?

  • t-test: find out if there is a significant difference between 2 levels of the independent variable

  • spearman’s rank: find out if there is a correlation between many levels of the independent variable

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79

what is a null hypothesis?

a hypothesis which states there is no significant difference between two variables and that the difference observed is purely due to chance or error

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