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Biology Alevel > Cloning And Biotechnology > Flashcards

Cloning And Biotechnology Flashcards

1
Q

What is vegetative reproduction and the vegetative organs of plants?

A

Reproduction from vegetative parts of plants - prodcue clones of parent
Vegetative organs:
Root/shoot tips
Axillary buds (leaves and stem meet)
Vascular cambium (xylem and phloem)

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2
Q

Methods of natural vegetative propagation?

A

Runners
Tubers
Rhizomes
Bubbles
Suckers
Offsets

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3
Q

What are runners?

A

horizontal stems that grow along soil surface away from parent plant (so no competition for water/nutrients) with NODES/STEM TIPS
- roots form under nodes forming new plant

E.g strawberries, peppermint, spider plants

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4
Q

What are tubers?

A

Form when tip of a stem become swollen with food, with buds on tuber surface that can develop into new shoots
E.g potatoes

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5
Q

What are bulbs?

A

Form when a leaf base becomes swollen with stored food and bud inside bulb can form new shoots

E.g daffodils

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6
Q

What are suckers?

A

Shoots that emerge from shallow root buds of parent plant
E.g elm trees

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7
Q

What are rhizomes?

A

Horizontal underground stems that store food/produce new vertical shoots/roots from buds on nodes along rhizome

E.g marram grass

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8
Q

Natural method of vegetative propagation:
PRACTICAL :producing cuttings FROM STEMS?

A
  1. Cut 5-10cm piece from end of parent plant’s stem using sharp,sterile tool
  2. Remove leaves from lower end of cutting, leaving just one at the tip
  3. Dip lower end of cutting in rooting powder —> contains hormones that induce root formation
  4. Plant your cutting in a pot containing suitable growth medium (well drained compost)
  5. Provide cutting with warm/moist environment - cover pot with plastic bag or put in a propagation
  6. Once it has formed its own roots, can plant it elsewhere to continue growth
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9
Q

How to take root and leaf cuttings?

A

Root: take section of root /make angled cut on one end + then treat as stem cutting

Leaf cuttings: remove entire leaf, score the veins,and place it in a growth medium with scored veins facing down

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10
Q

Other natural methods of vegetative propagation to produce clones?

A

Taking cuttings
Grafting( join shoot of one plant to growing stem and root of another plant)
Layering (bending stem of a growing plant downwards so it enters soil/grows into new plant)

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11
Q

What is tissue culture and microproagation?

A

Technique used to artificially clone— plants from existing plants
Micropropagation: using tissue culture to produce large numbers of new plants from meristem tissue taken from sample plant

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12
Q

Method of tissue culture?

A
  1. Small tissue samples (EXPLANTS) taken from original plant that will be cloned
    Cells from STEM and ROOTS are used - they’re MERISTEM CELLS so differentiate into any type of cell (totipotent)
  2. Cells are STERILISED to kill microorganisms bacteria/fungi compete for nutrients with plant cells - decreasing their growth rate
  3. Cell placed on culture medium contains plant nutrients (glucose ) and growth hormones (auxins)
  4. Cells in each explant divide to form undifferentiated mass of cells - CALLUS
  5. Callus cells transferred to new medium with specific conditions to encourage shoot/root formation - callus cells differentiate/develop into plantlets
  6. When roots/shoot of plantlets formed, they’re move to growth medium LIKE SOIL - can continue to grow - GENETICALLY IDENTICAL TO PARENT
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13
Q

Advantages of artificial plant cloning?

A

Rapid method of producing LARGE NO. new plants (rare and endangered species can be saved from extinction)
Plants can be cloned if sexual reproduction is not possible - e.g plants that are hard to grow from seed
Will all have desirable characteristic - resistance to pest/disease , high yield , colour etc.
plants are free from disease, as they use meristem as explant
Whole plants can be created from GM cells/tissues - retain GM characteristics

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14
Q

Disadvantages of artificial plant cloning?

A

Expensive /labour intensive process
Tissue culture is susceptible to microbial contamination
No genetic variation - all susceptible to same disease/environ factors (crops grown in monoculture allow rapid spread of disease/pests due to closeness of plants)

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15
Q

Ways to produce artificial clones of animals, by reproductive cloning?

A

Embryo twining
Somatic cell nuclear transfer (SCNT)

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16
Q

Process of embryo twinning?

A

Produced offspring that are clones of each other , NOT THEIR PARENTS
1. Zygote (fertilised egg) created by IVF
2. Zygote divides by mitosis to form embryo
3. Embryo is divided into 2 half embryos ad continues to divide
4. Each embryo placed into uterus of surrogate mother

(Cannot predict how many offspring will be produced) - guarantees desirable characterstics in offspring

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17
Q

What is SCNT?

A

Used successfully on DOLLY the sheep
1. Animal to be cloned donates body cell
2. Egg cell us extracted from egg donor and enucleated (nucleus removed by suction/discarded)
3. Nucleus from body cell injected into enucleated cell by ELECTRIC SHOCK
4. Shock triggers egg cell to divide by mitosis forming embryo
5. Embryo implanted into surrogate mother uterus

OFFSPRING WILL BE CLONE OF ORIGINAL ANIMAL

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18
Q

What is therapeutic cloning?

A

Technique that uses cloned cells to replace dead/damaged cell that cause loss of function in an individual
Embryos are cloned + subdivided
Each individual embryo cell is a TOTIPOTENT STEM CELL that can be cultured/artificially differentiated into any type of specialised cell

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19
Q

Use of therapeutic cloning?

A

Replacing specialised brain tissue in PARKINSON’S disease sufferers

20
Q

Arguments for animal cloning?

A

Animals with desirable characteristics/high yield can be cloned
Preserve endangered species
Produce regenerated organs - no risk of rejection in patient
Test medicinal drugs on cloned cells/tissues avoids animal/people testing

21
Q

Arguments against anima cloning?

A

Success rate of SCNT is low / expensive - cloned animals may be less healthy/abnormalities /have shorter life span
Ethical issues- is it right to create life (embryo) to destroy it?
Lack of genetic variation - all susceptible to same disease/pest

22
Q

Biotechnology?

A

Use of living organisms or parts of living organisms in industrial processes

23
Q

Why are microorganisms used in biotechnology?

A

Have simple growth requirements
Their food is cheap/readily available
Occupy little space
Reproduce quickly/short life cycle
Can be grown on industrial scale
Don’t have non-reproductive tissues/organs

PROVIDE MAT ECONOMIC ADVANATGES

24
Q

Applications of microbes in biotechnology?

A

Brewing - yeast ferment sugars anaerobically producing ethanol + CO2
Baking - CO2 produced in sugar fermentation by yeast helps bread dough rise
Cheese making - chymosin (enzyme from GM yeast), coagulates milk to curd, while bacteria and moulds contribute to acidification, thickening,flavour
Medicines - bioengineered fungi/bacteria produce drugs (antibiotic penicillin and insulin)
Bioremediation - microbes speed up degradation of pollutants - e.g oil spills
Single celled protein

25
Q

How are single celled proteins (mycoproteins) made by biotechnology?

A

Fusarium venenatum- fungus used
- used to make single celled/mycoproteins
E.g QUORN
Substitute for veg/healthy option for non-veg
- contains no animal fat/cholesterol

26
Q

Advantages of using microorganisms to make food?

A

Low fat, high protein products - mycoprotein
No animal welfare issues
Large amounts of land not required
Growth is not seasonal
Microorganisms can be grown on substrates that are waste products of other industries - SCP removes waste
Protein production many times faster than animal/plant protein

27
Q

Disadvantages of food production by microorganisms?

A
  • people may not want to eat fungal protein/food grown on waste
  • fermenter contaminated by other bacteria lead to ruined product
  • Sterile techniques needed to avoid contamination - EXPENSIVE
    Microbial biomass contains a lot of nucleic acids - not safe for human consumption so must be removed
    Protein doesn’t have same taste/texture as normal protein sources
    Bacteria can be infected by viruses - care taken to prevent this
28
Q

2 reasons to culture microorganisms?

A

Generate biomass of microorganisms - e.g to produce SCP as animal feedstock
Manufacture compounds the microbes synthesise (e.g antibiotics, vitamins,enzymes)

29
Q

What are primary and secondary metabolites?

A

Primary : substances produced in processes that are essential for normal microbial functioning
(E.g ethanol from anaerobic respiration in yeast

Secondary : substances produced in non-essential processes
(E.g antibiotics /plant defence chemicals)

30
Q

How are microbes cultured in the lab?

A
  1. Sterilise all equipment before use (e.g hold wire inoculating loop in Bunsen flame - flame creates convection current above bench, prevents contamination of any microbes in air /killed in flame)
  2. Dip sterilised inoculating loop into starter culture
  3. Transfer the microbes to a PETRI DISH containing sterile nutrient medium by zig-zagging loop across agar (lid should be opened as little as possible at side facing Bunsen - decrease risk of contamination)
  4. Close plates + tape them so not completely sealed (prevent growth of anaerobic microbes)
  5. Label plates (type of microbe, date and conditions)
  6. Incubate plates upside down (prevent condensation dropping onto agar) under required conditions (not above 25 degrees - less likely to grow harmful pathogens)
    7 repeat 1-6 for a control agar dish with no bacteria
  7. Assess microbial growth by observing colony formation on the agar
31
Q

Need for aseptic techniques and examples?

A

Ensue microbes being investigated dont escape or become contaminated with other microbes
E.g washing hands
No food/drink in lab
Disinfecting work surfaces with disinfectant/alcohol
Wear gloves/googles
Work close to Bunsen
Sterilise equipment

32
Q

2 methods to culture microorganisms by fermentation?

A

BATCH fermentation: microorganisms grown in batches in fermentation vessel until nutrients deplete/waste accumulates
After one culture cycle is complete, vessel is emptied/cleaned before starting next batch
KNOWN AS CLOSED CULTURE

CONTINUOUS fermentation: continually supply fresh nutrients/removing culture broth
- maintains growth of culture indefinitely

33
Q

Microbial growth curve in batch cultures?

A
  1. Lag phase- cells have slow initial growth as they adapt to environment +prodcue enzymes
  2. Log phase (exponential phase)- Rapid doubling of cell numbers occurs under ideal conditions, and growth rate is at its maximum.
  3. Stationary phase - Growth rate plateaus as nutrients diminish and waste accumulates
    cell growth = to cell death
  4. Death phase - Cell death rate > cell growth rate due to resource limitation and build up of toxins.
34
Q

Microbial growth curve in batch cultures?

A
  1. Lag phase- cells have slow initial growth as they adapt to environment +prodcue enzymes
  2. Log phase (exponential phase)- Rapid doubling of cell numbers occurs under ideal conditions, and growth rate is at its maximum.
  3. Stationary phase - Growth rate plateaus as nutrients diminish and waste accumulates
    cell growth = to cell death
  4. Death phase - Cell death rate > cell growth rate due to resource limitation and build up of toxins.
35
Q

How to maintain optimum conditions in fermenter?

A

Control:
PH: monitored by probes + acids/alkalis added to provide optimum pH for enzyme activity —> max rate of reaction +growth + productivity of microorganisms
Temperature:opt temps maintained using WATER JACKET - allows max enzyme acitvity/yield
O2 availability : sterile air pumped in proving optimum oxygen - max respiration /yeild
Nutrient supply : nutrients added/circulated for max yield
Agitation: paddles used to ensure even distribution of temp,nutrients,pH.O2 in fermenter
Contamination: cleaned between cultures (contamination can lead to competition for nutrients,O2,space —> reduce yield)
Waste removal : constantly removed to prevent toxic effects on microbes as affects their growth

36
Q

Factors affecting microbial growth and how can they be investigated (when growing microbes on agar)

A

Temperature - incubate 6 duplicate agar plates , keeping 3 at one temp and the other 3 at another temp- count no. Colonies (keeping lid on)
Calculate average no. Colonies formed + compare results
PH - add buffer solutions to marinating different pH levels
Nutrient availability -prepare agar with varying nutrient concs
Antimicrobial substances - add different antimicrobial compounds to agar plates

Can use turbidity as measure of growth instead of counting colonies

37
Q

How can populations of microorganisms be measured?

A

Direct counting - includes all cells (living or dead) and taking samples to count individual microorganisms
Viable counting - culture samples of microorganisms /counting colonies that grow (only use living cells)
Turbidity - measure living/dead microorganisms in SOLUTION - take absorbance reading using colorimeter
As microorganisms in culture reproduce/pop grows - suspension becomes more TUBID (cloudy)- colorimeter measure change in turbidity (by how much light can pass through suspension) at fixed time intervals
- Plot pop growth curve to show population growth over time

38
Q

Equation to calculate rate of cell division by BINARY FISSION?

A

N = No x 2^n

N = the final number of bacteria
N0 = the initial number of bacteria
n = the number of divisions

39
Q

Why are logarithmic scales used to show microbial growth?

A

Allow for wide range of values to be displayed on single graph
(Not equal intervals between numbers on y-axis)
- wide range of cell numbers fit onto same scale

40
Q

Why are serial dilutions used in growing microorganisms?

A

Once bacteria have been cultured in desired conditions, may be too many bacteria to count due to so many colonies or colonies OVERLAP —> form a lawn
- serial dilutions dilutes bacteria in broth before plating so solves this problem

41
Q

How are serial dilutions taken?

A

Taking a series of dilutions of a stock solution (sample of microorganism culture broth)
Conc decreases by same quantity between each test tube

42
Q

What are immobilised enzymes?

A

Enzyme that is attached to an insoluble material to prevent mixing with the product
Immobilised by being:
Attached to an inert substance
Enclosed in a capsule/matrix - entrapment
Contained within partially permeable membrane - encapsulation
Binding - to insoluble support materials (cellulose/collagen fibres by covalent/ionic bonds)

43
Q

Purpose of enzyme immobilisation? How do they work?

A

Used in industrial processes so enzyme can be REUSED in future processes
- also avoids need to separate enzyme from product later on

  1. Immobilised enzymes in a COLUMN
  2. Substrate runs through column and forms ENZYME-SUBSTRATE COMPLEXES + products are produced
  3. Products flow out column , leaving enzymes behind to catalyse reaction again
44
Q

Advantages of immobilised enzymes?

A

Cost effective- resuse enzymes - reduce need to buy new enzymes
Product purity- produces enzyme free products - avoiding contamination of product with enzyme
Improved stability - immobilised enzymes more tolerant of temp/pH changes than enzymes in solution —> more stable/less likely to denature

45
Q

Disadvantages of immobilised enzymes?

A

Higher initial costs - specialist expensive equipment needed /immobilised enzymes are expensive - not cost effective for smaller industries
Reduced enzyme activity - immobilisation can reduce enzyme efficiency
Tech problems - equipment is complex + prone to more tech problems

46
Q

How is lactase immobilised to produce lactose-free milk?

A

1.Lactase enzyme attached to ALGINATE beads to immobilise it
2.Lactase-containing beads packed into a column
3.Milk flows through column
4.Lactase hydrolyses lactose in milk into GLUCOSE + GALACTOSE- lactose free milk formed
5.Lactase remains in column , so more milk can be processed continually

Biology Alevel (28 decks)

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