Unit 8: Protoplast fusion - Practice Quiz

BTY559 — Biotechnology Laboratory-Ii 60 Questions
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1 What does the abbreviation PEG stand for in protoplast fusion?

fusion of protoplast by PEG method for hybrid production Easy
A. Phenyl ethyl glucoside
B. Potassium ethylene glycol
C. Polyethylene glycol
D. Polyester glycerol

2 A protoplast is a plant cell that lacks which structure?

fusion of protoplast by PEG method for hybrid production Easy
A. Cytoplasm
B. Cell wall
C. Nucleus
D. Plasma membrane

3 PEG acts as which type of agent in protoplast fusion?

fusion of protoplast by PEG method for hybrid production Easy
A. Nutrient
B. Antibiotic
C. Fusogen
D. Enzyme

4 The fusion of two protoplasts from different species produces a cell called a:

fusion of protoplast by PEG method for hybrid production Easy
A. Somatic hybrid
B. Gamete
C. Callus
D. Zygote

5 Which enzyme is commonly used to remove the cell wall and isolate protoplasts?

fusion of protoplast by PEG method for hybrid production Easy
A. Protease
B. Amylase
C. Lipase
D. Cellulase

6 Protoplast fusion is an example of which type of hybridization?

fusion of protoplast by PEG method for hybrid production Easy
A. Backcrossing
B. Sexual hybridization
C. Grafting
D. Somatic hybridization

7 In the PEG method, PEG promotes fusion mainly by inducing:

fusion of protoplast by PEG method for hybrid production Easy
A. Chlorophyll production
B. DNA replication
C. Cell wall synthesis
D. Membrane adhesion and agglutination

8 The main advantage of protoplast fusion over conventional breeding is that it overcomes:

fusion of protoplast by PEG method for hybrid production Easy
A. The need for water
B. Photosynthesis
C. Sexual incompatibility barriers
D. Enzyme activity

9 A hybrid cell containing the nucleus of one parent and the cytoplasm mixed from both is called a:

fusion of protoplast by PEG method for hybrid production Easy
A. Zygote
B. Cybrid
C. Spore
D. Clone

10 Which of the following is a chemical method of protoplast fusion?

fusion of protoplast by PEG method for hybrid production Easy
A. PEG-induced fusion
B. Micro-injection
C. Electrofusion
D. Sonication

11 During PEG treatment, protoplasts are usually kept in a medium that is:

fusion of protoplast by PEG method for hybrid production Easy
A. Completely dry
B. Strongly acidic
C. Highly hypotonic
D. Osmotically balanced (isotonic)

12 After fusion, protoplasts regenerate a new cell wall and then divide to form a:

fusion of protoplast by PEG method for hybrid production Easy
A. Pollen grain
B. Callus
C. Root hair
D. Seed

13 Which solution is often added after PEG treatment to help wash away PEG and improve fusion?

fusion of protoplast by PEG method for hybrid production Easy
A. High pH, high calcium solution
B. Dilute acid
C. Concentrated sugar syrup
D. Pure ethanol

14 Fusion of two protoplasts from the same species is called:

fusion of protoplast by PEG method for hybrid production Easy
A. Intraspecific fusion
B. Interkingdom fusion
C. Interspecific fusion
D. Intergeneric fusion

15 The first step in a protoplast fusion experiment is:

fusion of protoplast by PEG method for hybrid production Easy
A. Regeneration of plants
B. Selection of hybrids
C. Isolation of protoplasts
D. Callus formation

16 PEG used for protoplast fusion typically has a molecular weight in the range of:

fusion of protoplast by PEG method for hybrid production Easy
A. About 50000–100000
B. About 100–200
C. About 1500–6000
D. About 10–50

17 Which of the following enzymes helps degrade the middle lamella between plant cells during protoplast isolation?

fusion of protoplast by PEG method for hybrid production Easy
A. Pectinase
B. Ligase
C. Lipase
D. Cellulase

18 A somatic hybrid produced by protoplast fusion contains genetic material from:

fusion of protoplast by PEG method for hybrid production Easy
A. Only the cytoplasm
B. Only one parent
C. Neither parent
D. Both parental protoplasts

19 One disadvantage of the PEG fusion method is that PEG can be:

fusion of protoplast by PEG method for hybrid production Easy
A. Only used on animal cells
B. Unable to cause any adhesion
C. Toxic to protoplasts at high concentration
D. A source of nutrients

20 Protoplast fusion technology is mainly applied to produce improved:

fusion of protoplast by PEG method for hybrid production Easy
A. Plastic polymers
B. Crop plants with new traits
C. Mineral crystals
D. Synthetic fertilizers

21 In PEG-mediated protoplast fusion, what is the primary role of polyethylene glycol (PEG)?

fusion of protoplast by PEG method for hybrid production Medium
A. It supplies nutrients for hybrid cell division
B. It acts as a fusogen that promotes membrane agglutination and adhesion
C. It selectively kills non-fused parental protoplasts
D. It digests the cell wall to release protoplasts

22 Why is a high pH (--) high- solution often used together with PEG during fusion?

fusion of protoplast by PEG method for hybrid production Medium
A. It provides a carbon source for the protoplasts
B. It lowers the osmotic pressure of the medium
C. It removes residual cell wall fragments
D. It enhances membrane fusion after PEG-induced adhesion

23 A product formed by the fusion of two protoplasts from different species, containing nuclei of both parents, is called a:

fusion of protoplast by PEG method for hybrid production Medium
A. Homokaryon
B. Heterokaryon precursor
C. Somatic hybrid
D. Cybrid

24 A cybrid produced through protoplast fusion is characterized by:

fusion of protoplast by PEG method for hybrid production Medium
A. Nucleus of one parent and cytoplasm from both parents
B. Nuclei of both parents and cytoplasm of one parent
C. Nuclei and cytoplasm from a single parent
D. Complete fusion of both nuclei and both cytoplasms

25 Before PEG-mediated fusion, protoplasts must first be isolated. Which treatment achieves this?

fusion of protoplast by PEG method for hybrid production Medium
A. Centrifugation in a sucrose density gradient
B. PEG treatment at high concentration
C. Exposure to high pH calcium buffer
D. Enzymatic digestion of the cell wall using cellulase and pectinase

26 Which concentration range of PEG is typically effective for inducing protoplast fusion?

fusion of protoplast by PEG method for hybrid production Medium
A. About -- (w/v)
B. About -- (w/v)
C. About -- (w/v)
D. About -- (w/v)

27 One major advantage of the PEG method over electrofusion for protoplast fusion is that it:

fusion of protoplast by PEG method for hybrid production Medium
A. Requires no selection step to recover hybrids
B. Produces only heterokaryons with no homokaryons
C. Guarantees fusion between only two protoplasts
D. Is inexpensive and allows mass fusion of many protoplasts at once

28 During PEG fusion, protoplasts must be maintained in an osmotically balanced medium mainly to prevent:

fusion of protoplast by PEG method for hybrid production Medium
A. Loss of chloroplast pigments
B. PEG from degrading the plasma membrane
C. Bursting or shrinking of the wall-less protoplasts
D. Premature nuclear fusion

29 A researcher fuses protoplasts of two plants where one is resistant to a herbicide and the other to an antibiotic. Fused hybrids are selected on medium containing both agents. This selection strategy relies on:

fusion of protoplast by PEG method for hybrid production Medium
A. PEG selectively killing parental protoplasts
B. The higher growth rate of homokaryons
C. Random survival of parental cells
D. Complementation of two dominant selectable markers in the hybrid

30 The sequence of events in PEG-mediated fusion is best represented as:

fusion of protoplast by PEG method for hybrid production Medium
A. Membrane fusion → PEG treatment → agglutination → protoplast isolation
B. Protoplast isolation → PEG-induced agglutination → membrane fusion → hybrid selection
C. Hybrid selection → agglutination → PEG treatment → protoplast isolation
D. PEG treatment → protoplast isolation → hybrid selection → agglutination

31 Why is protoplast fusion particularly useful for producing hybrids between sexually incompatible plant species?

fusion of protoplast by PEG method for hybrid production Medium
A. It removes the need for chromosome pairing entirely
B. It doubles the seed set of both parents
C. It converts pollen into functional egg cells
D. It bypasses fertilization barriers by combining somatic cells directly

32 In PEG fusion, dimethyl sulfoxide (DMSO) is sometimes added to the PEG solution to:

fusion of protoplast by PEG method for hybrid production Medium
A. Neutralize the toxic effect of calcium ions
B. Digest the remaining cell wall
C. Act as the primary osmoticum
D. Increase fusion frequency by improving membrane fluidity

33 A product of fusion between two protoplasts of the SAME species/parent is termed a:

fusion of protoplast by PEG method for hybrid production Medium
A. Homokaryon
B. Heterokaryon
C. Cybrid
D. Somatic hybrid

34 Prolonged exposure to high concentrations of PEG during fusion mainly causes:

fusion of protoplast by PEG method for hybrid production Medium
A. Regeneration of the cell wall
B. Toxicity and death of the protoplasts
C. Increased chloroplast replication
D. Enhanced nuclear fusion efficiency

35 The intermediate cell containing two or more distinct nuclei immediately after membrane fusion but before nuclear fusion is a:

fusion of protoplast by PEG method for hybrid production Medium
A. Protoplast aggregate
B. Synkaryon
C. Heterokaryon
D. Cybrid

36 In a somatic hybridization experiment, if PEG treatment gives many fusion products but very few true hybrids survive on selection medium, the most likely explanation is:

fusion of protoplast by PEG method for hybrid production Medium
A. Nuclear fusion is impossible in PEG method
B. PEG destroyed all the selectable markers
C. The osmoticum was too concentrated for hybrids only
D. Most fusions are homokaryotic or non-viable, and only rare heterokaryons form viable hybrids

37 Which of the following is a limitation of the PEG method of protoplast fusion?

fusion of protoplast by PEG method for hybrid production Medium
A. It is non-selective and produces a mixture of fusion products
B. It requires expensive electrical equipment
C. It cannot fuse protoplasts of different species
D. It is unable to induce agglutination of protoplasts

38 After successful fusion and washing, the fusion products are cultured on nutrient medium primarily to allow:

fusion of protoplast by PEG method for hybrid production Medium
A. Re-isolation of fresh protoplasts
B. Digestion of the plasma membrane
C. Immediate PEG detoxification
D. Cell wall regeneration and division into hybrid callus

39 Somatic hybridization by PEG fusion is especially valuable for transferring which trait, often controlled by cytoplasmic genes?

fusion of protoplast by PEG method for hybrid production Medium
A. Cytoplasmic male sterility (CMS)
B. Seed color
C. Leaf shape
D. Flowering time only

40 The main reason a high-, high-pH wash follows PEG addition rather than being applied first is that:

fusion of protoplast by PEG method for hybrid production Medium
A. Calcium ions dissolve PEG completely
B. Fusion occurs before any adhesion of membranes
C. PEG must first bring protoplasts into close adhesion before fusion can be triggered
D. High pH is needed to isolate protoplasts from tissue

41 During PEG-mediated protoplast fusion, PEG promotes agglutination and membrane contact primarily because it acts as a molecular dehydrating agent. What is the direct physicochemical consequence of this dehydration at the membrane interface that permits fusion?

fusion of protoplast by PEG method for hybrid production Hard
A. Permanent covalent cross-linking of membrane glycoproteins between the two protoplasts
B. Removal of water molecules from the surface reduces the thickness of the hydration shell, allowing closer apposition of adjacent membranes
C. An increase in transmembrane potential that electroporates the lipid bilayer
D. Enzymatic digestion of the plasma membrane phospholipid bilayer exposing the cytoplasm

42 A researcher fuses protoplasts of species A () and species B (). Assuming a symmetric somatic hybrid retains both complete genomes without chromosome loss, what would be the chromosome number of the hybrid cell?

fusion of protoplast by PEG method for hybrid production Hard
A.
B.
C.
D.

43 Adding a high concentration of at an alkaline pH (around pH 9–10) to the PEG fusion protocol enhances fusion frequency. Which mechanism best explains this synergistic effect?

fusion of protoplast by PEG method for hybrid production Hard
A. bridges negatively charged membrane surfaces and high pH alters surface charge, together destabilizing membranes to favor fusion
B. High pH precipitates PEG, physically forcing protoplasts together into aggregates
C. activates cellulase enzymes that continue digesting residual cell wall material
D. enters the cytoplasm to trigger mitotic division immediately after contact

44 In a PEG fusion experiment, when two protoplasts fuse but their nuclei do NOT fuse and remain separate within a common cytoplasm, the resulting product is called a:

fusion of protoplast by PEG method for hybrid production Hard
A. Heterokaryon
B. Cybrid with no nuclear content
C. Homokaryon
D. Synkaryon

45 Why is PEG of molecular weight around 1500–6000 Da typically preferred over very low molecular weight PEG (e.g., 200 Da) for protoplast fusion?

fusion of protoplast by PEG method for hybrid production Hard
A. Low MW PEG cannot penetrate the intact cell wall of the protoplast
B. Higher MW PEG produces stronger dehydration and adhesion between membranes while causing tolerable toxicity
C. Higher MW PEG dissolves the plasma membrane more completely for fusion
D. Low MW PEG permanently binds nuclear DNA and blocks division

46 The stepwise dilution (gradual elution) of PEG with a fusion medium is a critical step. What is the primary rationale for diluting PEG slowly rather than washing it off abruptly?

fusion of protoplast by PEG method for hybrid production Hard
A. Abrupt washing chemically inactivates the PEG and prevents any fusion
B. Slow dilution allows time for the cell wall to regenerate before fusion
C. Rapid dilution causes PEG to polymerize and trap the protoplasts permanently
D. Gradual removal allows controlled rehydration and membrane reorganization that completes fusion while limiting osmotic shock

47 A major limitation of PEG-mediated fusion compared to electrofusion is its non-selectivity and low reproducibility. Which downstream requirement does this limitation make absolutely essential for recovering true hybrids?

fusion of protoplast by PEG method for hybrid production Hard
A. An efficient selection or screening system to distinguish hybrids from parental and homofused products
B. Complete removal of all cell wall material before culturing
C. Use of only haploid parental protoplasts to simplify analysis
D. Immediate transfer of fused products to soil for hardening

48 In somatic hybridization, if a protoplast fusion produces a cell containing the nucleus of one parent but organelles (chloroplasts/mitochondria) from both parents, the product is termed a:

fusion of protoplast by PEG method for hybrid production Hard
A. Symmetric hybrid
B. Allopolyploid
C. Cybrid
D. Synkaryon

49 Which combination of factors, if all present, would MOST likely reduce viable hybrid recovery in a PEG fusion protocol?

fusion of protoplast by PEG method for hybrid production Hard
A. Excessively high PEG concentration, prolonged exposure time, and impure PEG containing toxic contaminants
B. Use of freshly isolated, osmotically stabilized protoplasts
C. Moderate PEG concentration, brief exposure, and high-purity PEG
D. Presence of ions and slightly alkaline pH during elution

50 The classic Carlson somatic hybrid of two Nicotiana species was recovered by exploiting hormone-autotrophy of the hybrid. What principle of hybrid selection does this exemplify?

fusion of protoplast by PEG method for hybrid production Hard
A. Auxin overdose that kills only parental protoplasts
B. Visual chlorophyll deficiency selection based on color of colonies
C. Genetic complementation, where the hybrid gains a phenotype neither parent alone could sustain in culture
D. Antibiotic resistance conferred exclusively by PEG treatment

51 Suppose in a fusion mixture, the probability that any given protoplast is from species A is and from species B is , and pairwise fusions occur randomly. What fraction of all two-protoplast fusion events would be true A–B heterofusions?

fusion of protoplast by PEG method for hybrid production Hard
A.
B.
C.
D.

52 Why must protoplasts be maintained in an osmotically balanced (isotonic/slightly hypertonic) medium throughout the PEG fusion procedure?

fusion of protoplast by PEG method for hybrid production Hard
A. Osmotic balance is required for the cell wall to regenerate during fusion
B. Lacking a cell wall, protoplasts would burst under hypotonic conditions or shrink excessively under strong hypertonic stress
C. The osmoticum provides the carbon source for hybrid cell division
D. The osmoticum directly triggers membrane fusion independent of PEG

53 When distantly related (sexually incompatible) species are fused by PEG, hybrid cells frequently undergo progressive elimination of chromosomes of one parent during subsequent divisions. What is the most direct consequence for the resulting regenerated plant?

fusion of protoplast by PEG method for hybrid production Hard
A. It reverts entirely to one parental species with no genetic contribution from the other
B. It becomes an asymmetric hybrid retaining the full genome of one parent plus only part of the other's genome
C. It becomes a perfectly balanced symmetric hybrid identical to the fusion product
D. It always remains a stable heterokaryon that never divides

54 A common strategy uses irradiated 'donor' protoplasts and iodoacetate-treated 'recipient' protoplasts in PEG fusion (donor–recipient method). What is the combined logic of these two treatments?

fusion of protoplast by PEG method for hybrid production Hard
A. Both treatments increase the toxicity of PEG so fusion happens faster
B. Irradiation sterilizes contaminants and iodoacetate lowers the medium pH for fusion
C. Irradiation fragments donor DNA to force asymmetric transfer, while iodoacetate inactivation ensures only fusion products, not recipient parents, survive
D. Both treatments dissolve the cell membranes to guarantee 100% fusion

55 If the PEG solution used for fusion is prepared without any divalent cations and at neutral pH, and no cations are supplied during elution, the expected outcome is:

fusion of protoplast by PEG method for hybrid production Hard
A. Agglutination occurs but the frequency of actual membrane fusion is markedly reduced
B. The protoplasts immediately regenerate their cell walls
C. Protoplasts are completely protected and cannot aggregate at all
D. Fusion frequency reaches its theoretical maximum of 100%

56 Which statement correctly contrasts a key advantage of PEG fusion despite its drawbacks of toxicity and randomness?

fusion of protoplast by PEG method for hybrid production Hard
A. It guarantees survival of every fused product without any toxicity
B. It selectively fuses only genetically compatible protoplasts with high precision
C. It requires no specialized equipment and can fuse a large population of protoplasts simultaneously and inexpensively
D. It eliminates the need for any post-fusion selection or screening

57 A student obtains only of colonies as verified hybrids after PEG fusion despite good agglutination. Which analytical conclusion is most justified?

fusion of protoplast by PEG method for hybrid production Hard
A. The low yield proves that PEG cannot mediate fusion between these species
B. High agglutination does not guarantee high true-fusion or hybrid survival; homofusion and parental regrowth dominate without effective selection
C. The osmoticum concentration must have been too low, causing all hybrids to lyse
D. PEG molecular weight was too high, which prevented any membrane contact

58 In the fusion product, mitochondrial and chloroplast genomes from both parents initially coexist but often segregate or recombine over cell generations. This phenomenon is significant because it can generate:

fusion of protoplast by PEG method for hybrid production Hard
A. Identical nuclear genotypes in all daughter cells
B. Complete loss of all organellar DNA within one division
C. Novel cytoplasmic combinations such as recombinant mitochondrial genomes useful for traits like cytoplasmic male sterility
D. Permanent fixation of both parental organelle genomes in every cell forever

59 Consider a fusion protocol where exposure time to PEG is systematically varied: 5 min, 15 min, and 45 min. Fusion frequency rises then falls, peaking near 15 min. Which interpretation best explains this dose–response curve?

fusion of protoplast by PEG method for hybrid production Hard
A. PEG becomes more effective the longer it acts, and toxicity plays no role in the outcome
B. Shorter exposure is always superior because PEG never contributes to fusion after 5 min
C. Longer contact initially increases fusion, but excessive exposure raises cytotoxicity, killing potential fusants and lowering net yield
D. Fusion frequency should increase indefinitely with time, so the 45 min drop must be a measurement error

60 To confirm that a regenerated plant is a genuine somatic hybrid rather than a parental escape, which combination of evidence is MOST rigorous?

fusion of protoplast by PEG method for hybrid production Hard
A. Only the fact that the colony survived on selection medium
B. Only intermediate leaf morphology observed by eye
C. Only faster growth rate compared to either parent
D. Intermediate morphology plus molecular markers (e.g., isozymes, RFLP/PCR) and chromosome counts confirming both parental genomes