49. Marathon
® 1G, granular systemic insecticide.
50. Plant isolation device: plastic transparent floral sleeves (e.g.,
8454-T straight sleeve BOPP 60 Â 40 Â 15 cm, www.flo
wersleeves.com) for 11 cm diameter pots, or other devices for
plant isolation such as Aracons™ (Lehle Seeds and Arasystem,
www.arasystem.com) or pollination bags (e.g., 56 cm  76 cm
Delnet
® Apertured Films pollination bag, Schweitzer-Mauduit
International, www.swmintl.com©).
51. Hand sieve; Arabidopsis: 425 μm mesh screen or other Brassicaceae: 200 μm—3.35 mm mesh screen depending on species.
52. Small coin envelopes (e.g., 6 cm  9 cm) or small glass jars
(125 mL) or other containers.
53. Surface disinfectant.
2.2 Control
of Environmental
Growth Conditions
for Optimal Plant
Growth
1. Data loggers, e.g., HOBO
® U14 LCD.
2.3 Preparation
of Seeds for Shortand Long-Term
Storage
1. 2 mL polypropylene cryovials with threaded lids and gaskets or
other sealed containers for permanent seed storage.
2.4 Seed Quality
Control
1. Dissecting microscope or magnifying lenses.
2. Plastic Petri dishes (10 cm diameter) or other similar
containers.
3. Absorbent paper, e.g., filter paper 9 cm diameter.
4. Distilled water.
5. Parafilm
® or tape.
3 Methods
3.1 Growth
of Arabidopsis
and Other
Brassicaceae Plants
and Cultures
3.1.1 Liquid and Gas
Sterilization of Arabidopsis
Seeds for Growth
on Solid Media
Growth of Arabidopsis in experimental settings such as selection of
drug resistant and transformed plants, examination of early root
and shoot phenotypes, and identification of homozygous lethal
mutants is typically conducted in sterile conditions on solid
media. Since seeds are occasionally contaminated with microorganisms and dust, the use of an effective seed sterilization regime is
critical. There are two widely used methods of seed sterilization
using bleach or chlorine gas. Liquid bleach sterilization is a practical
method to sterilize a small number of Arabidopsis seeds at a time,
while large numbers of Arabidopsis seeds can be sterilized easily
using chlorine gas. The methods described have been developed for
Arabidopsis, but can be applied to related species.
6
Christopher Calhoun et al.
® 1G, granular systemic insecticide.
50. Plant isolation device: plastic transparent floral sleeves (e.g.,
8454-T straight sleeve BOPP 60 Â 40 Â 15 cm, www.flo
wersleeves.com) for 11 cm diameter pots, or other devices for
plant isolation such as Aracons™ (Lehle Seeds and Arasystem,
www.arasystem.com) or pollination bags (e.g., 56 cm  76 cm
Delnet
® Apertured Films pollination bag, Schweitzer-Mauduit
International, www.swmintl.com©).
51. Hand sieve; Arabidopsis: 425 μm mesh screen or other Brassicaceae: 200 μm—3.35 mm mesh screen depending on species.
52. Small coin envelopes (e.g., 6 cm  9 cm) or small glass jars
(125 mL) or other containers.
53. Surface disinfectant.
2.2 Control
of Environmental
Growth Conditions
for Optimal Plant
Growth
1. Data loggers, e.g., HOBO
® U14 LCD.
2.3 Preparation
of Seeds for Shortand Long-Term
Storage
1. 2 mL polypropylene cryovials with threaded lids and gaskets or
other sealed containers for permanent seed storage.
2.4 Seed Quality
Control
1. Dissecting microscope or magnifying lenses.
2. Plastic Petri dishes (10 cm diameter) or other similar
containers.
3. Absorbent paper, e.g., filter paper 9 cm diameter.
4. Distilled water.
5. Parafilm
® or tape.
3 Methods
3.1 Growth
of Arabidopsis
and Other
Brassicaceae Plants
and Cultures
3.1.1 Liquid and Gas
Sterilization of Arabidopsis
Seeds for Growth
on Solid Media
Growth of Arabidopsis in experimental settings such as selection of
drug resistant and transformed plants, examination of early root
and shoot phenotypes, and identification of homozygous lethal
mutants is typically conducted in sterile conditions on solid
media. Since seeds are occasionally contaminated with microorganisms and dust, the use of an effective seed sterilization regime is
critical. There are two widely used methods of seed sterilization
using bleach or chlorine gas. Liquid bleach sterilization is a practical
method to sterilize a small number of Arabidopsis seeds at a time,
while large numbers of Arabidopsis seeds can be sterilized easily
using chlorine gas. The methods described have been developed for
Arabidopsis, but can be applied to related species.
6
Christopher Calhoun et al.