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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.

Sharma, V.K.; Monostori, T.; Hause, B.; Maucher, H.; Göbel, C.; Hornung, E.; Hänsch, R.; Bittner, F.; Wasternack, C.; Feussner, I.; Mendel, R.R.; Schulze, J. Genetic transformation of barley to modify expression of a 13-lipoxygenase Acta Biol. Szeged 49, 33-34 , (2005)

Immature scutella of barley were transformed with cDNA coding for a 13-li-poxygenase of barley (LOX-100) via particle bombardment. Regenerated plants were tested by PAT-assay, Western-analysis and PCR-screening. Immunocytochemical assay of T0 plants showed expression of the LOX cDNA both in the chloroplasts and in the cytosol, depending on the presence of the chloroplast signal peptide sequences in the cDNA. A few transgenic plants containing higher amounts of LOX-derived products have been found. These are the candidates for further analysis concerning pathogen resistance.
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