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Plant Tissue Culture > Types of tissue culture > Flashcards

Types of tissue culture Flashcards

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1
Q

what are the eight types of plant tissue culture

A
  1. whole plant
  2. embryo
  3. root
  4. cell suspension
  5. protoplast
  6. callus
  7. microspore
  8. meristem
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2
Q

whole plant culture

A
  • maintains entire plant in sterile conditions
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3
Q

applications of whole plant culture

A
  1. arabidopsis - seed surface sterilized, sown on medium. water, support, minerals, and light are only requirements
  2. horticulture - some seeds only germinate in specific conditions
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4
Q

embryo culture

A
  • a type of whole plant culture, but with embryos
  • avoids dormancy problems
  • avoids embryo/endosperm incompatibility in hybrids
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5
Q

what is embryo rescue

A

obtain interspecific hybrid plants prior to abortion of embryos due to post-fertilization barriers

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6
Q

protoplast culture

A
  • cells are stripped of middle lamella and cell wall by pectinases and cellulases
  • protoplast remains alive
  • good for increasing plant ploidy and producing hybrids
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7
Q

protoplast culture specifications

A
  • medium requirements critical (bc of lack of cell walls)
  • protoplast vulnerable to osmotic concentrations
  • protoplast fusion requires media that enables cells to stick together
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8
Q

what is the difference between callus culture and liquid suspension culture?

A

callus culture - undifferentiated or unorganized mass of cells on agar
suspension culture - single or groups of cells are suspended

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9
Q

how is callus culture established?

A

a) diversity of cell types - less differentiated cells are more responsive to callus induction on simple media
b) physiological status of explant - nutrient status, hormonal content, dormancy status
c) genotype

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10
Q

Removed

A
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11
Q

callus culture features

A
  1. compact - cells tightly joined and the tissue mass is one solid piece
  2. friable - cells loosely joined and individual cells readily seperable
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12
Q

callus

A

a tissue that develops in response to injury caused by physical or chemical

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13
Q

callus forms due to:

A
  1. removal of cells within the explant from organizational controls
  2. provision of mineral nutrients and growth regulators for autonomous and indeterminate cell growth
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14
Q

callus culture growth patterns

A
  1. growth patterns leading to organized development - morphogenisis
  2. growth patterns leading to continued proliferation of unorganized callus -maintenance
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15
Q

callus culture- adventitious organogenesis and somatic embryogenesis

A

adventitious organogenesis - regeneration
somatic embryogenesis - development of embryos from somatic tissue

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16
Q

somatic embryogenesis

A

stimulation of callus or suspension cells to undergo a developmental pathway that mimics the development of zygotic embryo

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17
Q

zygotic embryogenesis

A

formation of an embryo as a result of double fertilization of the ovule

18
Q

microspore derived embryogenesis

A

production of embryos from microspores
- result requires haploid doubling

19
Q

meristem culture

A

meristem can produce an entire shoot and ultimately a whole plant

20
Q

micropropagation

A

creating multiple clones of a plant using plant tissue culture

21
Q

stages of microspore culture

A

i) inititation- explants placed in tissue medium for shoot inititaion
II) multiplication - cultures maintained on initiation medium for this stage (shoots multiply by 3X/month)
III) rooting - auxin only, focus is producing root tissue
IV) hardening off - acclimatizing plants to new conditions

22
Q

micropropagation advantages

A
  • from one to many plantlets rapidly
  • genetically identical clones
  • multiplication in controlled lab conditions
  • continuous propagation year round
  • potential for disease-free propagules
23
Q

micropropagation disadvantages

A
  • equipment/facility intensive operation
  • requires trained technicians
  • protocols not optimized for all species
  • propagules may be too expensive
24
Q

transient transformation

A
  • does not depend on integration into plant genome
  • fast
  • expression often disappears after less than a week
25
Q

stable transformation

A
  • integrated into plant genome
  • take longer
  • transgene passed to daughter cells
    -expression throughout plant
26
Q

transformation methods

A
  • agrobacterium
  • viruses
  • rhizobium
27
Q

agrobacterium transformation method

A
  • infects plant by inserting its DNA into plant DNA
  • host cells grow and divide rapidly
  • produce opine (carbon source and nitrogen source for bacteria)
28
Q

Agrobacterium advantages and disadvantages

A

advantages:
- more efficient than biolistics
- low # of insertion events/cell
disadvantages:
- doesnt work for all species
- insertion of bacterial DNA with GOI
- IP issues

29
Q

agrobacterium transformation steps

A

1) co-cultivation - bacteria infect plant tissue (plasmid DNA inserted into plant cells)
2) Kill phase - bacteria must be killed off, transferred to antibiotic media with high cytokinin conc.
3) regeneration phase - high BA content, cells with gene divide and become meristematic, must select for cells containing gene with antibiotic

30
Q

biolistics

A
  • gold particles coated with DNA
  • projected into plant cells
  • selection of transgenic cells
  • regeneration
31
Q

biolistics advantages vs disadvantages

A

advantages
- any tissue that can be regenerated through tissue culture can be transformed
- good for transient expression studies
disadvantages
- several copies of transgene insert into a chromosome (high copy events silenced)
- not as efficient as agrobacterium
- large DNA fragments may be damaged bc shearing

32
Q

CRISPR delivery

A

agrobacterium
- inserts into plant genome
- remove crispr genes by breeding with unaltered plants
protoplast
- deliver crispr enzymes and RNA guide molecules that dont insert into the genome
biolistics
- insert DNA, crispr enzymes and RNA
- if DNA is inserted into genome it must be removed

33
Q

events

A

insertion of a particular transgene into a specific location on a chromosome

34
Q

event: inserts into gene critical for survival or development

A

effect: knocks out gene and cell dies or cant divide

35
Q

event:inserts into locus that does not allow for expression

A

effect: never expressed transgene

36
Q

event: expressed in original transgenic plant but is silenced in later generations

A

effect: offspring doesnt express transgene

37
Q

event: inserts into gene critical for reproduction

A

effect: cannot pass event to offspring

38
Q

event:inserts into region genes not normally found or has an alteration in sequence

A

effect: expression occurs but not optimal level

39
Q

event: inserts in a gene that influences agronomic traits and yield

A

effect: yield drag

40
Q

why are desriable events rare?

A
  • transgene must insert in chromosome part that allows gene expression
  • cant disrupt other plant genes
  • most events never leave lab or greenhouse
  • others rejected in field
  • must produce many events to find a desirable one

Plant Tissue Culture (8 decks)

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