depend on this location (Fig. 1). It is important to choose the
right geographic orientation of the site, similar to that of
natural populations in the neighborhood of the selected site
[62]. Overall, natural populations of Arabidopsis do not tolerate open sites with strong direct sun during a long period of the
day. In addition, it is very convenient to flatten with a hoe the
land where pots or trays will be placed, and to dig out 5–10 cm
of the soil to remove all existing vegetation. This will reduce
heterogeneity in water availability and invasion of competitor
seeds, which will occur despite using pots or trays to grow
plants.
13. It is important to place the experiment in sites with good
natural drainage because Arabidopsis performs very badly in
waterlogged areas. In some cases, pots/trays can be placed on
platforms or tables to avoid waterlogging problems, although
it is always preferable to set the pots/trays at ground level to
make use of natural soil humidity.
14. When working in a new location for the very first time, it is
advisable to run a pilot experiment to detect those zones within
the facility affecting positively or negatively Arabidopsis plant
development. Although a pilot experiment in field conditions
requires a whole year, the success of the forthcoming experiments will strongly depend on this preliminary information
about the particularities of the experimental facility and the
actions that can be taken to reduce risks.
15. Many traits, but specially germination, seedling features, and
the subsequent phenology, are strongly affected by the environment of the mother plant [64]. To avoid such maternal
effects on your field phenotypes, it is strongly advisable that all
seeds to be grown in the same field experiment are first multiplied simultaneously in a single common experiment, with
minimum environmental variation. Such multiplication of
materials is usually done under controlled conditions, but it
can also be carried out in a field site [26, 35].
16. Seeds to be sown in each pot or tray should be prepared in
advance to have them ready on the planting date. For evolutionary studies estimating fitness components, it is particularly
important to know the exact number of seeds sown in pots or
trays because seedling establishment as rosettes and survival to
reproduction strongly depend on the initial number of seeds
sown. When sowing a small number of seeds per pot (<150),
these can be directly counted by eye or using a stereo microscope, and stored in Eppendorf tubes in darkness at room
temperature. Larger amounts of seeds can be weighed using
weight estimates for 100 seeds available for different Arabidopsis genotypes [65]. However, since seed weight depends on the
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right geographic orientation of the site, similar to that of
natural populations in the neighborhood of the selected site
[62]. Overall, natural populations of Arabidopsis do not tolerate open sites with strong direct sun during a long period of the
day. In addition, it is very convenient to flatten with a hoe the
land where pots or trays will be placed, and to dig out 5–10 cm
of the soil to remove all existing vegetation. This will reduce
heterogeneity in water availability and invasion of competitor
seeds, which will occur despite using pots or trays to grow
plants.
13. It is important to place the experiment in sites with good
natural drainage because Arabidopsis performs very badly in
waterlogged areas. In some cases, pots/trays can be placed on
platforms or tables to avoid waterlogging problems, although
it is always preferable to set the pots/trays at ground level to
make use of natural soil humidity.
14. When working in a new location for the very first time, it is
advisable to run a pilot experiment to detect those zones within
the facility affecting positively or negatively Arabidopsis plant
development. Although a pilot experiment in field conditions
requires a whole year, the success of the forthcoming experiments will strongly depend on this preliminary information
about the particularities of the experimental facility and the
actions that can be taken to reduce risks.
15. Many traits, but specially germination, seedling features, and
the subsequent phenology, are strongly affected by the environment of the mother plant [64]. To avoid such maternal
effects on your field phenotypes, it is strongly advisable that all
seeds to be grown in the same field experiment are first multiplied simultaneously in a single common experiment, with
minimum environmental variation. Such multiplication of
materials is usually done under controlled conditions, but it
can also be carried out in a field site [26, 35].
16. Seeds to be sown in each pot or tray should be prepared in
advance to have them ready on the planting date. For evolutionary studies estimating fitness components, it is particularly
important to know the exact number of seeds sown in pots or
trays because seedling establishment as rosettes and survival to
reproduction strongly depend on the initial number of seeds
sown. When sowing a small number of seeds per pot (<150),
these can be directly counted by eye or using a stereo microscope, and stored in Eppendorf tubes in darkness at room
temperature. Larger amounts of seeds can be weighed using
weight estimates for 100 seeds available for different Arabidopsis genotypes [65]. However, since seed weight depends on the
102
Carlos Alonso-Blanco et al.