Unit 2: Micropropagation; Somatic Hybridization - Practice Quiz

BTY540 — Plant Biotechnology 60 Questions
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1 Micropropagation refers to the in vitro propagation of plants using which technique?

Micropropagation and its stages Easy
A. Grafting on rootstock
B. Cutting and layering in the field
C. Seed germination in soil
D. Tissue culture

2 Which of the following is the correct first stage of micropropagation?

Micropropagation and its stages Easy
A. Multiplication of shoots
B. Hardening in the greenhouse
C. Rooting of shoots
D. Initiation of aseptic culture

3 The small piece of plant tissue used to start a micropropagation culture is called a(n):

Micropropagation and its stages Easy
A. Callus
B. Explant
C. Embryoid
D. Protoplast

4 The final stage of micropropagation, where plantlets are gradually adapted to external environmental conditions, is known as:

Micropropagation and its stages Easy
A. Callus induction
B. Multiplication
C. Initiation
D. Hardening (acclimatization)

5 Somatic embryogenesis is the process of forming embryos from:

somatic embryogenesis Easy
A. Pollen grains only
B. Fertilized egg cells
C. Somatic (vegetative) cells
D. Endosperm tissue only

6 Embryos produced through somatic embryogenesis are commonly referred to as:

somatic embryogenesis Easy
A. Protoplasts
B. Gametes
C. Zygotic embryos
D. Somatic embryos (embryoids)

7 A key feature that distinguishes a somatic embryo from a shoot bud is that a somatic embryo is:

somatic embryogenesis Easy
A. Bipolar with both root and shoot poles
B. Only capable of forming roots
C. Formed only after fertilization
D. Always larger than a zygotic embryo

8 Embryo culture involves the aseptic isolation and in vitro growth of:

embryo culture Easy
A. Mature seeds in soil
B. Pollen tubes
C. Root tips only
D. Immature or mature embryos

9 One common purpose of embryo culture is to:

embryo culture Easy
A. Overcome seed dormancy
B. Induce polyploidy in leaves
C. Produce protoplasts
D. Isolate chloroplasts

10 Embryo rescue is mainly used to save embryos that would otherwise:

embryo rescue Easy
A. Abort before maturity
B. Grow too rapidly
C. Produce excess seeds
D. Form only roots

11 Embryo rescue is especially valuable in producing which type of plants?

embryo rescue Easy
A. Interspecific or intergeneric hybrids
B. Haploid plants only
C. Pure inbred lines
D. Genetically identical clones

12 A major application of micropropagation is the rapid production of:

applications of micropropagation Easy
A. Sterile pollen grains
B. Large numbers of identical plants
C. Genetically diverse populations
D. New animal cell lines

13 Micropropagation is useful for producing plants that are free of which of the following?

applications of micropropagation Easy
A. Cell walls
B. Vascular tissue
C. Viruses and pathogens
D. Chlorophyll

14 A protoplast is a plant cell that lacks its:

Somatic Hybridization: protoplast isolation and fusion Easy
A. Cell wall
B. Nucleus
C. Cytoplasm
D. Plasma membrane

15 Which enzymes are commonly used to remove the plant cell wall during protoplast isolation?

Somatic Hybridization: protoplast isolation and fusion Easy
A. DNA polymerase and ligase
B. Trypsin and pepsin
C. Cellulase and pectinase
D. Amylase and lipase

16 Which chemical agent is widely used to induce protoplast fusion?

Somatic Hybridization: protoplast isolation and fusion Easy
A. Polyethylene glycol (PEG)
B. Sodium chloride
C. Glucose
D. Ethidium bromide

17 The method of fusing protoplasts using an electric field is called:

Somatic Hybridization: protoplast isolation and fusion Easy
A. Electroporation of plasmids
B. Electrolysis
C. Electrophoresis
D. Electrofusion

18 After protoplast fusion, why is selection of hybrid cells necessary?

selection of hybrid cells Easy
A. Hybrid cells cannot divide
B. All cells become identical hybrids
C. The fusion mixture contains parental and hybrid cells together
D. No cells survive the fusion process

19 Which of the following can be used as a visual marker for selecting fused hybrid cells?

selection of hybrid cells Easy
A. Differences in cell colour or fluorescence
B. Root length
C. Seed size
D. Cell wall thickness

20 A symmetric hybrid produced by somatic hybridization contains:

symmetric and asymmetric hybrids Easy
A. Only cytoplasmic genes from one parent
B. No nuclear genetic material
C. The nucleus of one parent only
D. Complete nuclear genomes of both parents

21 A tissue culture lab observes that shoots multiplied in vitro fail to survive when directly transplanted to the field. Which stage of micropropagation was most likely inadequate?

Micropropagation and its stages Medium
A. Stage IV (Hardening/acclimatization)
B. Stage I (Initiation)
C. Stage III (Rooting)
D. Stage II (Multiplication)

22 During the multiplication stage of micropropagation, a high cytokinin-to-auxin ratio in the medium primarily promotes which response?

Micropropagation and its stages Medium
A. Somatic embryo maturation
B. Shoot proliferation
C. Root initiation
D. Callus dedifferentiation

23 Which explant is generally preferred for establishing virus-free plants through micropropagation?

Micropropagation and its stages Medium
A. Fully differentiated root segment
B. Shoot apical meristem tip
C. Mature leaf disc
D. Nodal stem cutting with axillary buds

24 Which feature distinguishes a somatic embryo from an adventitious shoot arising in culture?

somatic embryogenesis Medium
A. Presence of both shoot and root poles (bipolar structure)
B. Absence of a suspensor at any stage
C. Requirement of high cytokinin for its formation
D. Development directly from the zygote

25 In carrot suspension cultures, transferring embryogenic cells to a medium with reduced auxin (2,4-D) typically triggers which event?

somatic embryogenesis Medium
A. Progression of proembryos into globular, heart and torpedo stages
B. Reversion of embryos back into disorganized callus
C. Immediate germination into mature seedlings
D. Formation of unipolar adventitious roots only

26 Somatic embryos are the biological basis of which agricultural product?

somatic embryogenesis Medium
A. Grafted rootstocks
B. Transgenic pollen
C. Monoclonal antibodies
D. Synthetic (artificial) seeds

27 A breeder cultures immature embryos on nutrient medium to overcome seed dormancy and shorten the breeding cycle. This technique is best described as:

embryo culture Medium
A. Embryo culture
B. Protoplast culture
C. Anther culture
D. Meristem culture

28 Why do very young (immature) excised embryos often require a more complex medium than mature embryos?

embryo culture Medium
A. They are heterotrophic and depend on complex nutrients and growth factors normally supplied by the endosperm
B. They contain chlorophyll and need only light
C. They germinate without any exogenous nutrients
D. They possess large food reserves and need no sugar

29 In a wide (interspecific) cross, fertilization succeeds but the hybrid embryo aborts due to endosperm failure. Which technique can recover a viable hybrid?

embryo rescue Medium
A. Protoplast fusion
B. Embryo rescue
C. Micrografting
D. Cold stratification of seeds

30 Embryo rescue is most valuable in plant breeding because it primarily helps overcome which barrier?

embryo rescue Medium
A. Pre-zygotic pollen–stigma incompatibility
B. Self-incompatibility during pollination
C. Male sterility
D. Post-zygotic incompatibility

31 Which of the following is the single most important advantage of micropropagation over conventional propagation for the commercial ornamental industry?

applications of micropropagation Medium
A. Introduction of new genes into the genome
B. Rapid, large-scale multiplication of genetically uniform (true-to-type) plants
C. Elimination of the need for any greenhouse space
D. Guaranteed production of somaclonal variants

32 The unexpected appearance of genetic variation among micropropagated plants, often associated with a prolonged callus phase, is called:

applications of micropropagation Medium
A. Vernalization
B. Somaclonal variation
C. Cytoplasmic inheritance
D. Transgressive segregation

33 Which application relies on micropropagation to preserve endangered or elite germplasm for long periods?

applications of micropropagation Medium
A. Grafting onto wild rootstocks
B. Seed banks for recalcitrant seeds stored at room temperature
C. In vitro conservation / cryopreservation of germplasm
D. Field gene banks in open plots only

34 Which enzyme combination is typically used to remove the plant cell wall and release protoplasts?

Somatic Hybridization: protoplast isolation and fusion Medium
A. Cellulase and pectinase
B. Restriction endonuclease and ligase
C. Amylase and lipase
D. DNase and RNase

35 To prevent osmotic bursting of freshly isolated protoplasts, the medium must be:

Somatic Hybridization: protoplast isolation and fusion Medium
A. Strongly hypotonic with pure distilled water
B. Osmotically balanced (hypertonic/isotonic) using mannitol or sorbitol
C. Free of any sugars or sugar alcohols
D. Highly acidic with pH below 3

36 Which agent is most commonly used as a chemical fusogen to induce protoplast fusion?

Somatic Hybridization: protoplast isolation and fusion Medium
A. Colchicine
B. 2,4-D
C. Polyethylene glycol (PEG)
D. Sodium alginate

37 Two parental protoplasts, one resistant to antibiotic A and the other resistant to antibiotic B, are fused. Growing the mixture on medium containing both antibiotics selects for which cells?

selection of hybrid cells Medium
A. Fused hybrid cells carrying both resistance traits
B. Cells that lost both resistance genes
C. Only unfused protoplasts
D. Only the first parental type

38 Which method allows physical selection of fusion products based on differences such as colour of chloroplasts versus colourless (albino) protoplasts?

selection of hybrid cells Medium
A. Auxotrophic complementation
B. Visual/mechanical selection under the microscope
C. Random regeneration without selection
D. Antibiotic double resistance

39 An asymmetric somatic hybrid differs from a symmetric hybrid mainly because:

symmetric and asymmetric hybrids Medium
A. It lacks any nuclear DNA from either parent
B. It contains complete diploid genomes from both parents
C. It contains the complete genome of one parent and only a partial genome of the other
D. It contains only cytoplasmic organelles and no nucleus

40 To create an asymmetric hybrid, the donor protoplast's nuclear genome is usually fragmented before fusion by treating it with:

symmetric and asymmetric hybrids Medium
A. Warm water at 40°C
B. Prolonged darkness
C. Gamma or X-ray irradiation
D. High concentration of sucrose

41 During Stage II (multiplication) of micropropagation, a culture that initially showed vigorous shoot proliferation begins producing shoots with translucent, water-soaked, thick appearance and poor rooting. The most likely biochemical basis for this disorder is:

Micropropagation and its stages Hard
A. Excessive cytokinin combined with high relative humidity and poor gas exchange causing hyperhydricity
B. Somaclonal variation arising from prolonged undifferentiated callus phase
C. Accumulation of phenolic exudates oxidizing the medium and browning tissue
D. Depletion of auxin leading to loss of apical dominance and callus formation

42 A protocol yields a multiplication rate of 4 shoots per explant every 4-week subculture. Starting with 5 explants, approximately how many shoots are theoretically available after 20 weeks (5 subcultures), assuming no losses?

Micropropagation and its stages Hard
A.
B.
C.
D.

43 In carrot somatic embryogenesis, cells maintained in high 2,4-D form proembryogenic masses but fail to progress to globular embryos. The essential manipulation to trigger embryo development is:

somatic embryogenesis Hard
A. Removing or drastically reducing 2,4-D to relieve auxin-mediated repression of embryo polarity
B. Raising sucrose to 8% to impose permanent osmotic stress
C. Increasing 2,4-D concentration to strengthen the embryogenic signal
D. Adding high cytokinin to induce shoot organogenesis instead

44 Which feature most reliably distinguishes a true somatic embryo from an adventitious shoot arising via organogenesis?

somatic embryogenesis Hard
A. Presence of a single apical meristem connected to maternal vascular tissue
B. Requirement of exogenous auxin for continued root formation
C. Bipolar structure with both shoot and root meristems and no vascular connection to parent tissue
D. Origin from multiple cells forming a meristemoid

45 Immature embryos cultured in vitro often require a more complex medium than mature embryos. The primary physiological reason is:

embryo culture Hard
A. Mature embryos possess larger cotyledons demanding higher mineral salts
B. Immature embryos have completed dormancy and need only water imbibition
C. Immature embryos are heterotrophic and cannot yet synthesize needed growth factors, requiring supplemented amino acids and organic supplements
D. Mature embryos require complex organic nitrogen to break physiological dormancy

46 The phenomenon where an excised immature embryo germinates directly into a seedling without passing through normal maturation and dormancy is termed:

embryo culture Hard
A. Precocious germination
B. Polyembryony
C. Recalcitrant dormancy
D. Somatic embryogenesis

47 In a wide cross between two species, fertilization succeeds but the hybrid embryo aborts by day 12 due to endosperm breakdown. The optimal embryo rescue strategy is:

embryo rescue Hard
A. Cross the hybrid back to a parent before embryo excision
B. Excise and culture the embryo (or ovule/ovary) before abortion on a nurse-endosperm or enriched medium
C. Apply colchicine to the developing seed to double chromosomes in situ
D. Wait until seed maturity then germinate the seed conventionally

48 Ovule culture and ovary culture are chosen over direct embryo excision in some interspecific crosses primarily because:

embryo rescue Hard
A. The embryos are too small or fragile to excise at the abortion stage, so surrounding tissue provides support
B. The ovary contributes maternal chromosomes needed for hybrid viability
C. Excised embryos always undergo somaclonal variation whereas ovules do not
D. Ovule culture eliminates the need for hormones in the medium

49 Meristem-tip culture is preferred for producing virus-free plants because:

applications of micropropagation Hard
A. The high auxin content of meristems degrades viral RNA
B. The apical meristem lacks vascular connections and viruses are excluded from rapidly dividing meristematic cells
C. Meristems contain high concentrations of antiviral phenolics
D. Meristems are surrounded by a callose barrier impermeable to all pathogens

50 A commercial lab notices increasing phenotypic variation in a clonally micropropagated ornamental after many subcultures. To minimize this problem while maintaining clonal fidelity, the best practice is:

applications of micropropagation Hard
A. Extend subculture intervals to allow more genetic recombination
B. Regenerate exclusively through indirect somatic embryogenesis
C. Propagate via axillary shoot proliferation and limit the number of subcycles rather than relying on callus/adventitious regeneration
D. Increase 2,4-D to accelerate callus-mediated multiplication

51 During enzymatic protoplast isolation, cellulase and macerozyme (pectinase) are used together. Their complementary roles are:

Somatic Hybridization: protoplast isolation and fusion Hard
A. Pectinase degrades cellulose while cellulase protects the protoplast
B. Cellulase separates cells and pectinase digests the plasma membrane
C. Both degrade cellulose but at different pH optima for redundancy
D. Cellulase degrades the cellulose wall while pectinase dissolves the middle lamella separating cells

52 In PEG-mediated versus electrofusion of protoplasts, a key advantage of electrofusion is:

Somatic Hybridization: protoplast isolation and fusion Hard
A. It permanently fuses nuclei without requiring membrane destabilization
B. It selectively fuses only genetically compatible protoplasts
C. It avoids the need for an osmoticum during fusion
D. It allows precise, synchronous fusion with less chemical toxicity by aligning cells via dielectrophoresis then applying a DC pulse

53 When two different protoplast populations are mixed and fused randomly, the fusion products include several types. The desired heterokaryon is:

Somatic Hybridization: protoplast isolation and fusion Hard
A. A protoplast that failed to fuse but survived
B. A fusion product containing nuclei from both parental species within a common cytoplasm
C. A cell in which both nuclei have already fused into one
D. A fusion product with two identical nuclei from the same parent

54 In complementation-based selection, two parental lines each carry a non-allelic recessive mutation blocking growth on selective medium. Only fusion hybrids grow because:

selection of hybrid cells Hard
A. Fusion activates silenced genes through epigenetic reprogramming
B. The hybrid cell doubles its ploidy, masking both mutations
C. Each parental genome supplies the wild-type allele the other lacks, restoring the complete pathway in the hybrid
D. The selective medium kills all unfused protoplasts by toxicity

55 A researcher fuses protoplasts where one parent is chlorophyll-deficient (albino, cannot photosynthesize) and the other is sensitive to a metabolic inhibitor. Hybrids are selected on inhibitor-containing medium under light. This dual/complementation scheme works because:

selection of hybrid cells Hard
A. Only hybrids combine inhibitor resistance from one parent with photosynthetic competence from the other
B. The albino parent's chloroplasts dominate and neutralize the inhibitor
C. The inhibitor converts albino cells into photosynthetic ones
D. Light selectively lyses all non-hybrid protoplasts

56 To transfer only a small segment of the donor genome (e.g., a disease-resistance gene) into a recipient while retaining the recipient's genome intact, one produces an asymmetric hybrid by:

symmetric and asymmetric hybrids Hard
A. Doubling the donor chromosome number with colchicine before fusion
B. Irradiating the donor protoplasts (X/gamma rays) to fragment its chromosomes before fusion
C. Fusing equal, undamaged protoplasts from both parents
D. Treating recipient protoplasts with iodoacetate to inactivate its cytoplasm

57 A symmetric somatic hybrid between two distantly related species is often unstable and shows chromosome elimination. The main reason symmetric hybridization is limited for wide crosses is:

symmetric and asymmetric hybrids Hard
A. Symmetric fusion prevents any organelle mixing, causing lethality
B. The combined genome is always exactly diploid and thus fully fertile
C. Symmetric hybrids cannot regenerate into whole plants under any condition
D. Incompatibility between divergent genomes leads to spontaneous, uneven loss of chromosomes and sterility

58 A cybrid is best defined as a cell/plant containing:

cybrids Hard
A. The nucleus of one parent with cytoplasm (organelles) derived from both or the other parent
B. Doubled chromosome sets from a single parent only
C. Only chloroplasts and no functional nucleus
D. Nuclei of both parents but organelles of only one

59 To generate a cybrid transferring cytoplasmic male sterility (CMS) into an elite line, the standard donor–recipient protocol involves:

cybrids Hard
A. Doubling chromosomes of both parents with colchicine before fusion
B. Irradiating (enucleating) the CMS donor protoplasts and treating recipient protoplasts with iodoacetate, then fusing so the recipient nucleus combines with donor cytoplasm
C. Isolating mitochondria alone and microinjecting them into zygotes
D. Fusing two undamaged protoplasts and selecting the tallest regenerants

60 Fluorescence-activated cell sorting (FACS) can isolate heterokaryons when the two parental protoplasts are pre-labelled with different fluorochromes. The principle enabling selection is:

selection of hybrid cells Hard
A. Unfused protoplasts lose all fluorescence upon fusion
B. The dyes chemically fuse the membranes during sorting
C. Heterokaryons are heavier and separate purely by size
D. Only heterokaryons emit both fluorescence signals simultaneously, allowing dual-fluorescence gating