Unit 6: Recent Advancements in Plant Biotechnology; Molecular Markers - Practice Quiz

BTY540 — Plant Biotechnology 60 Questions
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1 What does the abbreviation TCL stand for in plant biotechnology?

thin cell layer technology (TCL) Easy
A. Thin Cell Layer
B. Tissue Culture Line
C. Total Cell Line
D. Thick Callus Layer

2 A transverse thin cell layer (tTCL) explant is cut in which orientation relative to the plant axis?

thin cell layer technology (TCL) Easy
A. At a 45-degree diagonal
B. Along the root tip only
C. Parallel to the axis
D. Perpendicular to the axis

3 What is the main advantage of using thin cell layer technology in micropropagation?

thin cell layer technology (TCL) Easy
A. High regeneration from small explant tissue
B. Prevention of all genetic variation
C. Elimination of the need for light
D. Complete avoidance of growth regulators

4 Which nanoparticles are most commonly used as antimicrobial/sterilizing agents in plant tissue culture?

nanoparticles as sterilizing agent and media components Easy
A. Silica nanoparticles for coloring
B. Gold nanoparticles for staining
C. Carbon nanotubes
D. Silver nanoparticles (AgNPs)

5 Nanoparticles added to culture media can act as a source of which type of nutrient?

nanoparticles as sterilizing agent and media components Easy
A. Only agar gelling agents
B. Only carbohydrates
C. Micronutrients / trace elements
D. Only vitamins

6 One key property of nanoparticles that makes them effective in tissue culture is their:

nanoparticles as sterilizing agent and media components Easy
A. Very high surface-area-to-volume ratio
B. Complete chemical inertness
C. Extremely large particle size
D. Inability to enter plant cells

7 Meta-topolin belongs to which class of plant growth regulators?

noble plant growth regulators (metatopolin, melatonin) Easy
A. Auxins
B. Abscisic acid
C. Cytokinins
D. Gibberellins

8 Meta-topolin is often preferred over BAP (6-benzylaminopurine) because it:

noble plant growth regulators (metatopolin, melatonin) Easy
A. Acts only as a herbicide
B. Reduces physiological disorders like hyperhydricity
C. Prevents root and shoot growth
D. Completely stops shoot formation

9 Melatonin in plants primarily functions as a:

noble plant growth regulators (metatopolin, melatonin) Easy
A. Antioxidant and growth regulator
B. Gelling agent for media
C. Restriction enzyme
D. Selective antibiotic

10 Melatonin is known to help plants tolerate which of the following?

noble plant growth regulators (metatopolin, melatonin) Easy
A. Only mechanical wounding
B. Only viral infection
C. Only nutrient excess
D. Various abiotic stresses

11 What does RFLP stand for?

RFLP Easy
A. Restriction Fragment Length Polymorphism
B. Restriction Fragment Ligation Product
C. Random Fragment Linkage Polymorphism
D. Repeated Fragment Length Pattern

12 RFLP analysis relies mainly on the use of which enzymes to cut DNA?

RFLP Easy
A. Restriction endonucleases
B. Helicases
C. Ligases
D. DNA polymerases

13 RAPD markers are generated using PCR with which type of primers?

RAPD Easy
A. Short random primers
B. Long sequence-specific primers
C. Fluorescent probes only
D. Restriction enzymes

14 A major limitation of RAPD markers is that they are:

RAPD Easy
A. Unable to use PCR
B. Dominant and less reproducible
C. Based only on RNA
D. Codominant and highly reproducible

15 SSR markers are also commonly known as:

SSR/ISSR Easy
A. Microsatellites
B. Telomeres
C. Centromeres
D. Minisatellites only

16 What does the abbreviation ISSR stand for?

SSR/ISSR Easy
A. Inter-Simple Sequence Repeat
B. Internal Single Sequence Region
C. Inverted Simple Sequence Reader
D. Isolated Short Sequence Repeat

17 AFLP technique combines which two key steps?

AFLP Easy
A. Only random priming
B. Sequencing and cloning only
C. Only ligation and staining
D. Restriction digestion and selective PCR amplification

18 SCAR markers are usually developed from which type of markers?

SCAR Easy
A. Isozyme markers
B. RFLP blots only
C. Protein markers
D. RAPD markers

19 Molecular markers are widely used in tissue culture to detect:

applications of molecular markers in plant biotechnology and tissue culture Easy
A. The pH of agar only
B. The color of the culture medium
C. The temperature of the incubator
D. Somaclonal variation and genetic fidelity

20 In plant breeding, molecular markers are commonly used for:

applications of molecular markers in plant biotechnology and tissue culture Easy
A. Spraying pesticides
B. Measuring soil color
C. Watering the crops
D. Marker-assisted selection (MAS)

21 A researcher excises a longitudinal strip from a tobacco stem containing only the epidermal and a few sub-epidermal cell layers. This preparation is most accurately described as which type of explant?

thin cell layer technology (TCL) Medium
A. Nodal segment explant
B. Callus-derived protoplast
C. Transverse thin cell layer (tTCL)
D. Longitudinal thin cell layer (lTCL)

22 Why does thin cell layer (TCL) technology generally give higher morphogenic efficiency than conventional large explants?

thin cell layer technology (TCL) Medium
A. TCL explants avoid the need for any plant growth regulators
B. The reduced number of cell layers minimizes internal gradients so cells respond more uniformly to exogenous growth regulators and nutrients
C. Thicker explants prevent contamination better
D. The larger tissue mass stores more endogenous auxin

23 In a TCL system, the developmental pathway (root, shoot, flower or callus) that the explant follows is primarily controlled by which factor?

thin cell layer technology (TCL) Medium
A. The ratio and concentration of plant growth regulators in the medium
B. The colour of the culture vessel
C. The genus of the agar supplier
D. The presence of activated charcoal only

24 Silver nanoparticles (AgNPs) are increasingly used as surface sterilants in plant tissue culture instead of mercuric chloride mainly because they:

nanoparticles as sterilizing agent and media components Medium
A. Permanently mutate the explant genome for resistance
B. Increase the sucrose content of the medium
C. Replace the need for a laminar air flow cabinet
D. Provide broad antimicrobial action with lower phytotoxicity and reduced environmental hazard

25 The antimicrobial activity of silver nanoparticles is best attributed to which mechanism?

nanoparticles as sterilizing agent and media components Medium
A. Release of Ag ions that damage microbial membrane proteins and generate reactive oxygen species
B. Physical blockage of stomata on the microbes
C. Lowering the osmotic potential of the agar
D. Chelation of sucrose in the medium

26 When added at low concentrations as media components, nanoparticles such as ZnO or TiO can enhance in vitro growth mainly by:

nanoparticles as sterilizing agent and media components Medium
A. Replacing agar as the gelling agent
B. Eliminating the requirement for light
C. Acting as micronutrient sources and modulating antioxidant and enzyme activity
D. Serving as the sole carbon source

27 Meta-topolin is often preferred over the classical cytokinin BAP (6-benzylaminopurine) in micropropagation because it:

noble plant growth regulators (metatopolin, melatonin) Medium
A. Completely eliminates the need for any auxin in rooting
B. Reduces shoot-tip necrosis and hyperhydricity while improving rooting after transfer
C. Functions primarily as an auxin rather than a cytokinin
D. Is a gaseous regulator applied only through the vessel headspace

28 Chemically, meta-topolin belongs to which class of cytokinins?

noble plant growth regulators (metatopolin, melatonin) Medium
A. Gibberellin-type diterpenes
B. Isoprenoid (zeatin-type) cytokinins
C. Aromatic (hydroxylated topolin) cytokinins
D. Auxin-type indole compounds

29 Melatonin (N-acetyl-5-methoxytryptamine) added to culture media is valued mainly for which combined roles?

noble plant growth regulators (metatopolin, melatonin) Medium
A. Functioning strictly as a herbicide
B. Acting as a potent antioxidant and modulating rooting and stress tolerance
C. Serving as the principal gelling agent
D. Providing the only nitrogen source in the medium

30 Melatonin is biosynthetically derived from which amino acid, sharing part of its pathway with auxin?

noble plant growth regulators (metatopolin, melatonin) Medium
A. Methionine
B. Arginine
C. Tryptophan
D. Glutamate

31 In an RFLP analysis, a point mutation abolishes a restriction site in one allele. What band pattern change is expected relative to the wild-type allele on a Southern blot?

RFLP Medium
A. Two smaller fragments replace one larger fragment
B. No change because RFLP detects only insertions
C. A single larger fragment replaces two smaller fragments
D. The DNA fails to hybridize with any probe

32 Which feature makes RFLP markers co-dominant, allowing heterozygotes to be distinguished from homozygotes?

RFLP Medium
A. Only the dominant allele generates a band
B. Amplification uses a single arbitrary primer
C. Both alleles produce their own detectable fragments simultaneously
D. The marker relies on random priming of anonymous loci

33 A key limitation of RAPD markers compared with most other PCR-based markers is that they are:

RAPD Medium
A. Dominant and often poorly reproducible across laboratories
B. Dependent on prior sequence and Southern blotting
C. Co-dominant and highly reproducible
D. Restricted to detecting methylation changes only

34 RAPD analysis differs fundamentally from targeted PCR because it uses:

RAPD Medium
A. Two long locus-specific primers designed from known sequence
B. Fluorescently labelled microsatellite-flanking primers
C. A single short primer of arbitrary sequence to amplify anonymous genomic regions
D. Restriction enzymes followed by ligation of adaptors

35 Simple sequence repeats (SSRs) are highly informative markers largely because their polymorphism arises from:

SSR/ISSR Medium
A. Random arbitrary primer binding
B. Variation in the number of tandem repeat units due to replication slippage
C. Loss of restriction sites from point mutations
D. Differential DNA methylation of promoters

36 How does ISSR primer design differ from SSR (microsatellite) primer design?

SSR/ISSR Medium
A. ISSR primers are arbitrary decamers identical to RAPD primers
B. ISSR uses restriction digestion instead of PCR
C. ISSR primers must be designed from cloned flanking sequences of each locus
D. ISSR primers are anchored microsatellite sequences that amplify regions between two repeats without needing flanking sequence data

37 Arrange the core steps of AFLP in the correct order:

AFLP Medium
A. Microsatellite priming → digestion → ligation → electrophoresis
B. Restriction digestion → adaptor ligation → pre-selective and selective PCR → gel resolution
C. Adaptor ligation → arbitrary priming → cloning → sequencing
D. PCR amplification → restriction digestion → Southern blot → hybridization

38 Why does AFLP achieve high multiplex ratios (many markers per reaction) with good reproducibility?

AFLP Medium
A. It uses arbitrary decamer primers at low stringency
B. It relies on hybridization of radioactive probes to blots
C. It amplifies only a single microsatellite locus at a time
D. Selective primers with defined 3′ bases combined with stringent PCR sample only a reproducible subset of restriction fragments

39 A SCAR marker is typically developed by:

SCAR Medium
A. Sequencing the ends of a useful RAPD/AFLP band and designing longer locus-specific primers
B. Digesting genomic DNA with rare cutters and ligating adaptors
C. Using an arbitrary decamer at low annealing temperature
D. Measuring repeat-number variation at microsatellites

40 Compared with the RAPD marker it is derived from, a SCAR marker generally shows:

SCAR Medium
A. Lower annealing temperature requirements
B. Greater reproducibility and locus specificity
C. Complete loss of any diagnostic value
D. Dependence on random arbitrary primers

41 A researcher isolates a transverse thin cell layer (tTCL) versus a longitudinal thin cell layer (lTCL) from the same tobacco floral stem. Which statement best explains why tTCL and lTCL frequently yield different morphogenic outcomes despite originating from the same explant?

thin cell layer technology (TCL) Hard
A. lTCL always produces roots while tTCL always produces shoots regardless of hormonal balance
B. tTCL cells are inherently totipotent while lTCL cells are permanently differentiated and cannot respond to growth regulators
C. The difference arises only from the surface area exposed to the medium and has no relation to tissue composition
D. tTCL contains a heterogeneous mix of tissue types across the cut plane, exposing multiple competent cell populations to inductive signals, whereas lTCL isolates a more homogeneous single-tissue strip

42 Why does TCL technology often achieve higher regeneration frequencies and greater control over morphogenesis compared to conventional explants?

thin cell layer technology (TCL) Hard
A. The small size prevents any contamination, eliminating the need for surface sterilization steps
B. TCL explants contain more chloroplasts, providing extra energy for organogenesis
C. Minimal tissue volume reduces correlative inhibition and endogenous hormone gradients, so exogenous regulators dominate the developmental fate of the few target cells
D. Reduced cell number increases somaclonal variation, which improves regeneration

43 In tissue culture, silver nanoparticles (AgNPs) are used both as sterilants and as media additives. What is the primary mechanistic reason AgNPs can simultaneously reduce microbial contamination and, at low doses, enhance shoot proliferation?

nanoparticles as sterilizing agent and media components Hard
A. AgNPs physically block bacterial cell division and simultaneously supply silver as an essential plant macronutrient
B. Controlled release of ions disrupts microbial membranes and enzymes while at low concentrations modulating ethylene signaling in plant tissues
C. AgNPs reflect UV light, sterilizing the medium while providing extra photosynthetic energy
D. AgNPs chelate all heavy metals, preventing both microbial growth and plant toxicity entirely

44 A researcher observes that increasing AgNP concentration in the medium initially boosts callus growth but sharply inhibits it beyond a threshold. Which explanation best accounts for this biphasic (hormetic) response?

nanoparticles as sterilizing agent and media components Hard
A. Nanoparticles aggregate only at low concentrations, becoming inert at high concentrations
B. Low doses are sterile and high doses introduce contamination that kills the callus
C. Low doses trigger mild oxidative signaling that stimulates growth, while high doses cause excessive ROS accumulation and oxidative damage overwhelming antioxidant defenses
D. The medium pH rises linearly with AgNP concentration, and only high pH is toxic

45 Meta-topolin (mT) is increasingly preferred over BAP (6-benzylaminopurine) for micropropagation of several species. Which combination of properties best explains mT's advantage in reducing physiological disorders during repeated subculture?

noble plant growth regulators (metatopolin, melatonin) Hard
A. mT is a synthetic auxin that suppresses all cytokinin activity, preventing shoot proliferation disorders
B. mT is metabolically inert and simply prevents any hormone from acting
C. mT irreversibly binds cytokinin receptors, providing a permanent single burst of activity
D. mT forms O-glucoside storage conjugates that are reversibly hydrolyzed, giving balanced cytokinin activity and reducing hyperhydricity and rooting inhibition seen with BAP

46 Melatonin (N-acetyl-5-methoxytryptamine) is used in plant tissue culture beyond a growth-regulatory role. Which dual functional property makes it particularly valuable for micropropagation under stress conditions?

noble plant growth regulators (metatopolin, melatonin) Hard
A. It acts as a potent free-radical scavenger and also influences auxin-like rooting and morphogenic responses
B. It functions only as a carbon source substituting for sucrose in the medium
C. It solely sterilizes the medium by killing endophytic bacteria
D. It is exclusively a rooting hormone with no antioxidant capacity

47 A plant geneticist scores an RFLP locus and finds that a homozygote shows a single band while the heterozygote shows three bands. What is the most likely molecular explanation for the extra band in the heterozygote?

RFLP Hard
A. A restriction site polymorphism where one allele retains an internal cut site producing two fragments while the other allele loses it, so the heterozygote shows both allelic patterns
B. The heterozygote has undergone a chromosome duplication tripling the copy number
C. PCR amplification artifact producing random extra fragments in heterozygotes only
D. RFLP always produces three bands due to probe cross-hybridization

48 Compared with PCR-based markers, why is classical RFLP considered highly reproducible across laboratories yet impractical for high-throughput screening?

RFLP Hard
A. It detects single-base changes by sequencing, which is cheap but non-reproducible
B. It relies on locus-specific hybridization independent of thermal cycling conditions, but requires large quantities of high-quality DNA, radioactive/labeled probes, and is labor-intensive
C. It amplifies microsatellites requiring only tiny DNA amounts but expensive fluorescent detection
D. It uses arbitrary primers that anneal at low stringency, making it fast but unreliable

49 RAPD markers are dominant and often criticized for poor reproducibility. Which single change in reaction conditions is MOST likely to alter the RAPD banding pattern and why?

RAPD Hard
A. A change in gel running voltage, because it alters primer annealing during PCR
B. A change in annealing temperature, because short arbitrary primers bind at low stringency and small temperature shifts change which mismatched priming sites amplify
C. A change in the loading dye color, because it affects primer specificity
D. A change in DNA quantity within a 10-fold range, because RAPD requires exact template amounts to bind probes

50 Two individuals share a RAPD band of identical size. Why is it risky to assume this shared band represents the same allele or locus (homology)?

RAPD Hard
A. Comigrating fragments of equal size may arise from different genomic regions or different priming events, so band identity does not guarantee sequence or locus homology
B. RAPD bands of equal size are always from the same locus due to primer specificity
C. Equal-sized bands indicate heterozygosity at a codominant locus
D. Shared bands prove the individuals are clonal because RAPD detects methylation

51 SSR (microsatellite) markers are prized as codominant and highly polymorphic. What molecular mechanism primarily generates the length polymorphism (variable repeat number) at SSR loci?

SSR/ISSR Hard
A. DNA polymerase slippage (replication slippage) during replication of the tandem repeats, causing gain or loss of repeat units
B. Point mutations converting one base to another within a single repeat
C. Random primer mismatches during low-stringency PCR
D. Restriction enzyme digestion cutting the repeats into variable fragments

52 ISSR and SSR markers both target microsatellites, yet ISSR does not require prior sequence knowledge whereas SSR does. Which statement correctly contrasts their primer design and dominance behavior?

SSR/ISSR Hard
A. ISSR uses single primers anchored to the repeat itself to amplify inter-repeat regions and is largely dominant, while SSR uses locus-specific flanking primers and is codominant
B. Both require restriction digestion before amplification and score as dominant markers
C. ISSR uses locus-specific primers and is codominant; SSR uses arbitrary primers and is dominant
D. Both use flanking locus-specific primers and are codominant, differing only in fragment size

53 In AFLP, selective amplification uses primers with selective nucleotides at their 3' ends. What is the functional purpose of adding these selective bases?

AFLP Hard
A. To reduce the enormous number of restriction fragments to a manageable subset by amplifying only those whose adjacent bases match the selective nucleotides
B. To convert the dominant markers into codominant markers
C. To add restriction sites so fragments can be re-digested for scoring
D. To label fragments radioactively for Southern hybridization

54 AFLP typically uses a rare cutter (e.g., ) combined with a frequent cutter (e.g., ). Why is this two-enzyme combination advantageous over using two rare cutters or two frequent cutters?

AFLP Hard
A. It generates predominantly one rare-frequent cutter fragment type of optimal size range for PCR and gel resolution, balancing fragment number and length
B. It ensures every fragment has two identical ends for efficient self-ligation
C. It allows detection of methylation at all cytosines in the genome
D. It eliminates the need for selective amplification primers

55 A RAPD band linked to a disease-resistance trait is converted into a SCAR marker. What is the principal benefit gained by this conversion?

SCAR Hard
A. SCAR uses shorter arbitrary primers, increasing the number of amplified loci
B. SCAR uses longer, locus-specific primers designed from the sequenced RAPD fragment, giving high reproducibility and often codominant scoring
C. SCAR abolishes the linkage between the marker and the trait, allowing independent inheritance
D. SCAR requires restriction digestion of the whole genome, improving polymorphism

56 A SCAR marker developed from a RAPD band amplifies a product in both resistant and susceptible plants of equal size, unlike the original RAPD which was present only in resistant plants. Which explanation best resolves this apparent loss of polymorphism?

SCAR Hard
A. The polymorphism in the original RAPD lay in a priming site outside the sequenced fragment ends, so the internal SCAR primers amplify both alleles and the polymorphism must be recovered via a restriction digest (CAPS) or primer redesign
B. The SCAR primers are too short to distinguish alleles, so no polymorphism can ever be recovered
C. SCAR markers by definition never show polymorphism and are used only for fingerprinting
D. The resistant and susceptible plants became genetically identical during marker conversion

57 To confirm the genetic fidelity (trueness-to-type) of micropropagated clones and detect somaclonal variation, which marker strategy provides the most robust genome-wide assessment, and why?

applications of molecular markers in plant biotechnology and tissue culture Hard
A. Using multiple marker systems together (e.g., ISSR + SSR + SCoT/AFLP) because each samples different genomic regions and mutation types, maximizing the chance of detecting variation
B. Using morphological observation alone, because it directly reflects all genetic changes
C. Using a single RAPD primer, because one arbitrary primer covers the entire genome
D. Using only RFLP with one probe, because Southern blotting scans the whole genome at once

58 In marker-assisted selection (MAS) for introgressing a resistance gene, why are flanking codominant markers on both sides of the target gene preferred over a single dominant marker within the gene region?

applications of molecular markers in plant biotechnology and tissue culture Hard
A. Flanking markers allow selection against recombinants and reduce linkage drag while codominance distinguishes heterozygotes from homozygotes, improving selection accuracy
B. Flanking markers are unnecessary because a within-gene marker never recombines away
C. Dominant single markers allow scoring of homozygotes versus heterozygotes directly
D. A single dominant marker gives more polymorphism and always eliminates linkage drag

59 A breeder wants to detect off-type contaminants and verify hybrid purity in a commercial seed lot. Which property of SSR markers makes them especially suited for this quality-control task?

applications of molecular markers in plant biotechnology and tissue culture Hard
A. Their codominance and multiallelic nature allow unambiguous identification of the expected heterozygous genotype and detection of selfed or contaminant seeds
B. Their low polymorphism ensures all seeds look identical, confirming purity
C. Their dominant inheritance masks heterozygotes, simplifying purity testing
D. Their reliance on random primers gives species-wide uniform patterns

60 When constructing a saturated linkage map for QTL analysis, why might a researcher favor AFLP markers in the early stage despite their dominant nature?

applications of molecular markers in plant biotechnology and tissue culture Hard
A. AFLP requires no restriction enzymes, making it the cheapest per-locus method
B. AFLP markers are all codominant, providing full zygosity information immediately
C. AFLP generates a very large number of polymorphic loci per reaction without prior sequence information, rapidly saturating the map, after which key regions can be converted to codominant markers
D. AFLP amplifies only single loci, ensuring each band maps to one gene