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Gasperini, D.; Acosta, I. F.; Farmer, E. E. Cotyledon Wounding of Arabidopsis Seedlings Bio Protoc 6, e1712, (2016) DOI: 10.21769/BioProtoc.1712
Damage to plant organs through both biotic and
abiotic injury is very common in nature. Arabidopsis thaliana 5-day-old
(5-do) seedlings represent an excellent system in which to study plant
responses to mechanical wounding, both at the site of the damage and in
distal unharmed tissues. Seedlings of wild type, transgenic or mutant
lines subjected to single or repetitive cotyledon wounding can be used
to quantify morphological alterations (e.g., root length, Gasperini et
al., 2015), analyze the dynamics of reporter genes in vivo (Larrieu et
al., 2015; Gasperini et al., 2015), follow transcriptional changes by
quantitative RT-PCR (Acosta et al., 2013; Gasperini et al., 2015) or
examine additional aspects of the wound response with a plethora of
downstream procedures. Here we illustrate how to rapidly and reliably
wound cotyledons of young seedlings, and show the behavior of two
promoters driving the expression of β-glucuronidase (GUS) in entire
seedlings and in the primary root meristem, following single or
repetitive cotyledon wounding respectively. We describe two procedures
that can be easily adapted to specific experimental needs.