3.2 Plant Material
for Establishment of In
Vitro Cultures
3.2.1 Potato
1. Sprout potato tubers in the darkness at room temperature to
achieve long nodal segments.
2. Cut potato shoots, remove leaves, cut nodal segments into
pieces of 2–3 cm, and continue with surface sterilization.
3.2.2 Mint
1. Harvest plant shoots from plants grown in the greenhouse
before flowering.
2. Remove leaves and cut nodal segments into pieces of 2–3 cm
and continue with surface sterilization.
3.2.3 Garlic Bulbs
1. Harvest garlic bulbs at maturity (see Fig. 2d) June–July in the
Northern Hemisphere.
2. Dry the bulbs using efficient ventilation to avoid molding.
Clean and remove soil when bulbs are dry (see Notes 4 and 5).
3. Store the bulbs using efficient ventilation at 4–10
C until use.
4. Before use, separate the cloves and remove the outer scale.
5. Remove the outer part of the thick storage scale and trim the
basal inner part to a cube of 10 mm size containing the basal
plate and the meristem.
6. Rinse this part under running tap water for few seconds and
continue with surface sterilization.
3.2.4 Garlic Bulbils
1. Harvest garlic bulbils at maturity (see Fig. 2h), dry and clean
them, and store them between 4 and 10
C until use (see Notes
4 and 5).
2. Remove the outer scale of the bulbils and sterilize them.
3.2.5 Garlic
Inflorescences
1. Harvest unripe inflorescences (see Fig. 2g) between April
and July in the Northern Hemisphere and store them in a
refrigerator at 4
C for 4–6 weeks (see Notes 4 and 5).
2. Cut the inflorescences, when the spatha is still closed and
sterilize them.
3.2.6 Shallots
1. Harvest shallots at maturity (see Fig. 2k, July in the Northern
Hemisphere), dry and clean them, and store them at
4–10
C until use . Use for cryopreservation between January
and April.
2. Remove the outer and inner scale of the shallots. Trim the basal
inner part to a cube of 10 mm size containing the basal plate
and the meristems. Sterilize them.
3.3 Surface
Sterilization
1. Prepare sodium hypochlorite solution and add a drop of
Tween 20.
652
Angelika Senula and Manuela Nagel
for Establishment of In
Vitro Cultures
3.2.1 Potato
1. Sprout potato tubers in the darkness at room temperature to
achieve long nodal segments.
2. Cut potato shoots, remove leaves, cut nodal segments into
pieces of 2–3 cm, and continue with surface sterilization.
3.2.2 Mint
1. Harvest plant shoots from plants grown in the greenhouse
before flowering.
2. Remove leaves and cut nodal segments into pieces of 2–3 cm
and continue with surface sterilization.
3.2.3 Garlic Bulbs
1. Harvest garlic bulbs at maturity (see Fig. 2d) June–July in the
Northern Hemisphere.
2. Dry the bulbs using efficient ventilation to avoid molding.
Clean and remove soil when bulbs are dry (see Notes 4 and 5).
3. Store the bulbs using efficient ventilation at 4–10
C until use.
4. Before use, separate the cloves and remove the outer scale.
5. Remove the outer part of the thick storage scale and trim the
basal inner part to a cube of 10 mm size containing the basal
plate and the meristem.
6. Rinse this part under running tap water for few seconds and
continue with surface sterilization.
3.2.4 Garlic Bulbils
1. Harvest garlic bulbils at maturity (see Fig. 2h), dry and clean
them, and store them between 4 and 10
C until use (see Notes
4 and 5).
2. Remove the outer scale of the bulbils and sterilize them.
3.2.5 Garlic
Inflorescences
1. Harvest unripe inflorescences (see Fig. 2g) between April
and July in the Northern Hemisphere and store them in a
refrigerator at 4
C for 4–6 weeks (see Notes 4 and 5).
2. Cut the inflorescences, when the spatha is still closed and
sterilize them.
3.2.6 Shallots
1. Harvest shallots at maturity (see Fig. 2k, July in the Northern
Hemisphere), dry and clean them, and store them at
4–10
C until use . Use for cryopreservation between January
and April.
2. Remove the outer and inner scale of the shallots. Trim the basal
inner part to a cube of 10 mm size containing the basal plate
and the meristems. Sterilize them.
3.3 Surface
Sterilization
1. Prepare sodium hypochlorite solution and add a drop of
Tween 20.
652
Angelika Senula and Manuela Nagel