Techniques

Question 1

what are 6 types of contaminants

Answer 1

1. Bacteria - usually introduced withe explant
2. Fungi, yeast - enter on explant or air
3. Insects - thrips, mites
4. Initial contaminants - introduced with explant when disinfestation is insufficient
5. Persistant or latent contamination - usually bacterial, show endogenously in plant tissue
6. Introduced contamination - from poor technique

Question 2

Aseptic technique

Answer 2

without infection, avoiding contamination
- long hair tied back
- gloves, 70% ethanol
- no talking
- dont lean over work
- work in middle of hood
- dont disturb the air

Question 3

laminar flow hood

Answer 3

• removes particles from air
• air goes through HEPA filter

Question 4

surface sterilization

Answer 4

1. alcohol (phytotoxic)
2. Bleach (10-20 mins)
3. Rinsing (sterile water 3X, 5 mins)

Question 5

explant

Answer 5

piece of plant that initiates tissue culture process

Question 6

callus

Answer 6

unorganized/undifferentiated mass of cells

Question 7

subculture

Answer 7

to remove a growing culture from a vessel to another

Question 8

accession

Answer 8

bring new plant material into culture

Question 9

sources of explants

Answer 9

immature embryos, cotylendonary petioles, shoots, roots, stems

Question 10

what can antibiotics be used for

Answer 10

• to cleanup contaminated tissue stocks or remove microorganisms used for transformation
• Agrobacterium tumefaciens - used to insert transgenic plasmids into plant cell
• timentin