The WHIRLY (WHY) DNA/RNA binding proteins fulfil multiple but poorly characterised functions in leaf development. WHY1 transcript levels were highest in the bases of 7-day old barley leaves. Immunogold labelling revealed that the WHY1 protein was more abundant in the nuclei than the proplastids of the leaf bases. Transcript and metabolite profiling analysis of barley lines (W1-1 and W1-7) lacking WHY1, which show delayed greening compared to the wild type. While the transcript profile of leaf development was largely unchanged in W1-1 and W1-7 leaves, there were differences in levels of several transcripts encoding transcription factors associated with chloroplast development. These include a barley homologue of the Arabidopsis GATA transcription factor that regulates stomatal development, greening and chloroplast development, NAC1, two transcripts with similarity to Arabidopsis GLK1 and two transcripts encoding ARF transcriptions factors with functions in leaf morphogenesis and development. Chloroplast proteins were less abundant in the W1-1 and W1-7 leaves than the wildtype. The levels of TCA cycle metabolites and GABA were significantly lower in WHY1 knockdown leaves than the wild type. We conclude that WHY1 functions in the nuclei of the cells in the leaf bases contributes to the regulation of chloroplast development.

Current crop protection strategies against the fungal pathogen Botrytis cinerea rely on a combination of conventional fungicides and host genetic resistance. However, due to pathogen evolution and legislation in the use of fungicides, these strategies are not sufficient to protect plants against this pathogen. Defence elicitors can stimulate plant defence mechanisms through a phenomenon known as priming. Priming results in a faster and/or stronger expression of resistance upon pathogen recognition by the host. This work aims to study priming of a commercial formulation of the elicitor chitosan. Treatments with chitosan result in induced resistance in solanaceous and brassicaceous plants. In tomato plants, enhanced resistance has been linked with priming of callose deposition and accumulation of the plant hormone jasmonic acid (JA). Large-scale transcriptomic analysis revealed that chitosan primes gene expression at early time-points after infection. In addition, two novel tomato genes with a characteristic priming profile were identified, Avr9/Cf-9 rapidly-elicited protein 75 (ACRE75) and 180 (ACRE180). Transient and stable overexpression of ACRE75, ACRE180 and their Nicotiana benthamiana homologs, revealed that they are positive regulators of plant resistance against B. cinerea. This provides valuable information in the search for strategies to protect Solanaceae plants against B. cinerea.