21. Leaves and debris fallen from neighboring trees or brought by
the wind will often cover some pots during fall and winter
seasons. The effect of these leaves will be very much reduced
by covering the pots with a wire mesh (Fig. 1b, k). This mesh
will also reduce the impact of raindrops on pots/trays and will
avoid attacks by small mammals and birds.
22. The periodicity of the visits will depend on the location of the
field site and the goals of the experiment. Heavy rains can occur
occasionally in some areas, which might disturb part of the
experiment. Very cold regions (sites in northern latitudes or
high altitudes) can be affected by large periods of snow, thus
limiting the access to the site. It is convenient to register in a
field notebook the overall state of the site, the problems
encountered or any relevant incidence in every visit, including
photographs of the experimental field and detailed photographs of plants, pots and trays. The information recorded
throughout the experiment can be very important to interpret
some of the results or any incidence occurred during the development of the experiment.
23. It is advisable to record the number of seedlings grown per pot
or tray after the first rain period to know the number of seeds
remaining in each pot, which might germinate later in the
season (see Notes 1, 8, 9, and 16). Image analysis of pot
photographs can help scoring seedlings, rosette survival, and
early growth phenotypes [67]. Alternatively, seedlings,
rosettes, and adult reproductive plants can be counted in the
field directly with a hand counter. Mortality of the seedlings
and plants before reproduction will vary during winter depending on the local climate, the specific weather during the experiment, and the genotype, but survival to reproduction will
provide a major component of fitness.
24. Depending on the purpose of the experiment, plants might be
watered periodically, occasionally, or never. No watering
should be applied when minimal human disturbance is desired,
but soft watering during specific unexpected drought periods
might avoid full mortality during extreme dry episodes.
25. Exceptionally, specific pesticides can be used in case of uncommon massive infections or insect attacks, although that will
affect the interpretation of the results (see Note 7). The use
of pesticides should also comply with the actual legislation.
26. Some phenotypes can be scored by bringing the plants back to
the greenhouse (e.g., vernalization response; [63]).
27. Single plant fitness or fitness at the pot level can be estimated as
the product of plant survival to reproduction and fecundity
[27, 31, 39, 68]. Measurements of plant survival require knowing the precise number of starting seeds or seedlings (see Notes
104
Carlos Alonso-Blanco et al.
the wind will often cover some pots during fall and winter
seasons. The effect of these leaves will be very much reduced
by covering the pots with a wire mesh (Fig. 1b, k). This mesh
will also reduce the impact of raindrops on pots/trays and will
avoid attacks by small mammals and birds.
22. The periodicity of the visits will depend on the location of the
field site and the goals of the experiment. Heavy rains can occur
occasionally in some areas, which might disturb part of the
experiment. Very cold regions (sites in northern latitudes or
high altitudes) can be affected by large periods of snow, thus
limiting the access to the site. It is convenient to register in a
field notebook the overall state of the site, the problems
encountered or any relevant incidence in every visit, including
photographs of the experimental field and detailed photographs of plants, pots and trays. The information recorded
throughout the experiment can be very important to interpret
some of the results or any incidence occurred during the development of the experiment.
23. It is advisable to record the number of seedlings grown per pot
or tray after the first rain period to know the number of seeds
remaining in each pot, which might germinate later in the
season (see Notes 1, 8, 9, and 16). Image analysis of pot
photographs can help scoring seedlings, rosette survival, and
early growth phenotypes [67]. Alternatively, seedlings,
rosettes, and adult reproductive plants can be counted in the
field directly with a hand counter. Mortality of the seedlings
and plants before reproduction will vary during winter depending on the local climate, the specific weather during the experiment, and the genotype, but survival to reproduction will
provide a major component of fitness.
24. Depending on the purpose of the experiment, plants might be
watered periodically, occasionally, or never. No watering
should be applied when minimal human disturbance is desired,
but soft watering during specific unexpected drought periods
might avoid full mortality during extreme dry episodes.
25. Exceptionally, specific pesticides can be used in case of uncommon massive infections or insect attacks, although that will
affect the interpretation of the results (see Note 7). The use
of pesticides should also comply with the actual legislation.
26. Some phenotypes can be scored by bringing the plants back to
the greenhouse (e.g., vernalization response; [63]).
27. Single plant fitness or fitness at the pot level can be estimated as
the product of plant survival to reproduction and fecundity
[27, 31, 39, 68]. Measurements of plant survival require knowing the precise number of starting seeds or seedlings (see Notes
104
Carlos Alonso-Blanco et al.