Measuring pollination and seed set
At the end of each sampling day, we collected one style from a flower on
twelve pre-marked C. quamash plants and mounted styles on
fuchsin-tinted gelatin slides (Kearns & Inouye 1993). We counted the
number of conspecific and heterospecific pollen grains on stigmas using
a compound light microscope (Nikon Eclipse 80i, Nikon Instruments Inc.).
Seventy-two hours following pollinator observations and after the
initiation of pollen tube growth, we collected a second style from these
same pre-marked plants into 70% ethanol. In the lab, we softened styles
with 8M NaOH at 35ยบ C for 1 hour and stained pollen tubes by placing
softened styles in a solution of 0.05% aniline blue in 0.1M
KH2PO4 for 24 hours. We squashed styles
beneath cover slips on microscope slides and counted pollen tubes near
the base of styles using epifluorescence microscopy (Nikon Eclipse 80i,
Nikon Instruments Inc.). Two weeks after conducting pollinator
observations we collected fruits and scored ovules as fertilized or
unfertilized.