such as flowering time. Therefore, the protocols described here
should be taken only as a guide for the experimental setup and
design.
This chapter will address: (1) the growth of Arabidopsis plants
in a variety of environmental settings including growth chambers
and greenhouses, as well as in vitro, (2) critical and optimal conditions to grow healthy Arabidopsis plants, including quality control
measures, (3) seed harvest, preservation, and quality control,
(4) genetic crosses, and (5) transformation with Agrobacterium
tumefaciens (Agrobacterium). Significant emphasis is placed on
the equipment required for controlling and monitoring environmental conditions during plant growth. Plant and seed management protocols are given in chronological order.
2 Materials
2.1 Sterilization, Cell
Culture, Plant Growth,
and Seed Harvest
1. Arabidopsis seeds can be obtained from the public stock centers: Arabidopsis Biological Resource Center (ABRC, abrc.osu.
edu), Nottingham Arabidopsis Stock Centre (NASC, www.
arabidopsis.info), RIKEN BioResource Center (RIKEN BRC,
www.en.brc.riken.jp), French National Institute for Agricultural Research (INRA, www.inrgv.toulouse.inra.fr), other laboratory sources [1], and private sources such as Lehle Seeds
(www.arabidopsis.com).
2. Seeds of other members of the Brassicaceae can be obtained
from public stock centers: National Plant Germplasm System
(NPGS, www.ars-grin.gov), UK Vegetable Genebank (www.
warwick.ac.uk/fac/sci/lifesci/wcc/gru/genebank/), Tohoku
Univ. Brassica Seed Bank (http://www.agri.tohoku.ac.jp/
pbreed/Seed_Stock_DB/Stock_English_top.html),
European Search Catalogue for Plant Genetic Resources
(EURISCO, www.eurisco.ipk-gatersleben.de), and other laboratory sources [1].
3. Sterile plastic Petri dishes (10 cm or 15 cm diameter).
4. Murashige and Skoog Basal Salt Mixture (MS).
5. Sucrose.
6. Potassium hydroxide (KOH).
7. Distilled water.
8. Magnetic stirring device.
9. Beakers (1 L).
10. Glass bottles (1 L).
11. pH meter.
12. Microcentrifuge tubes.
13. Disposable Pasteur pipettes.
4
Christopher Calhoun et al.
should be taken only as a guide for the experimental setup and
design.
This chapter will address: (1) the growth of Arabidopsis plants
in a variety of environmental settings including growth chambers
and greenhouses, as well as in vitro, (2) critical and optimal conditions to grow healthy Arabidopsis plants, including quality control
measures, (3) seed harvest, preservation, and quality control,
(4) genetic crosses, and (5) transformation with Agrobacterium
tumefaciens (Agrobacterium). Significant emphasis is placed on
the equipment required for controlling and monitoring environmental conditions during plant growth. Plant and seed management protocols are given in chronological order.
2 Materials
2.1 Sterilization, Cell
Culture, Plant Growth,
and Seed Harvest
1. Arabidopsis seeds can be obtained from the public stock centers: Arabidopsis Biological Resource Center (ABRC, abrc.osu.
edu), Nottingham Arabidopsis Stock Centre (NASC, www.
arabidopsis.info), RIKEN BioResource Center (RIKEN BRC,
www.en.brc.riken.jp), French National Institute for Agricultural Research (INRA, www.inrgv.toulouse.inra.fr), other laboratory sources [1], and private sources such as Lehle Seeds
(www.arabidopsis.com).
2. Seeds of other members of the Brassicaceae can be obtained
from public stock centers: National Plant Germplasm System
(NPGS, www.ars-grin.gov), UK Vegetable Genebank (www.
warwick.ac.uk/fac/sci/lifesci/wcc/gru/genebank/), Tohoku
Univ. Brassica Seed Bank (http://www.agri.tohoku.ac.jp/
pbreed/Seed_Stock_DB/Stock_English_top.html),
European Search Catalogue for Plant Genetic Resources
(EURISCO, www.eurisco.ipk-gatersleben.de), and other laboratory sources [1].
3. Sterile plastic Petri dishes (10 cm or 15 cm diameter).
4. Murashige and Skoog Basal Salt Mixture (MS).
5. Sucrose.
6. Potassium hydroxide (KOH).
7. Distilled water.
8. Magnetic stirring device.
9. Beakers (1 L).
10. Glass bottles (1 L).
11. pH meter.
12. Microcentrifuge tubes.
13. Disposable Pasteur pipettes.
4
Christopher Calhoun et al.