Seed Preparation
Stems were stored in a cool, dry environment for no more than 90 days
before we manually separated seeds by type. Chasmogamous seeds were
shaken from inflorescences and manually extracted with forceps when
necessary. We tested three approaches to separating cleistogamous seeds.
Most were manually extracted from stems by carefully slicing the stems
so as not to damage seeds before removing them either individually or as
a group when possible. However, we also soaked a random subset of stems
containing both cleistogamous and chasmogamous seeds in room temperature
tap water to facilitate cleistogamous seed extraction and sorting with a
light table, hereafter referred to as soaked seeds. We compared these to
an additional randomized subset of cleistogamous seeds that were still
enclosed within the stem but separated into individual seed units,
further facilitating seed processing, hereafter referred to as sheathed
seeds. Once we extracted all seeds, we randomly weighed a subset of
non-soaked or sheathed chasmogamous and cleistogamous seeds
(~20 each) from each maternal plant to calculate average
seed weights by maternal plant.