In plants, a variety of stimuli trigger long-range calcium signals that travel rapidly along the vasculature to distal tissues via poorly understood mechanisms. Here, we use quantitative imaging and analysis to demonstrate that traveling calcium waves are mediated by diffusion and bulk flow of amino acid chemical messengers. We propose that wounding triggers release of amino acids that diffuse locally through the apoplast, activating the calcium-permeable channel GLUTAMATE RECEPTOR-LIKE 3.3 as they pass. Over long distances through the vasculature, the wound-triggered dynamics of a fluorescent tracer show that calcium waves are likely driven by bulk flow of a channel-activating chemical. We observed that multiple stimuli trigger calcium waves with similar dynamics, but calcium waves alone cannot initiate all systemic defense responses, suggesting that mobile chemical messengers are a core component of complex systemic signaling in plants.

Zea mays and Miscanthus × giganteus use NADP-ME subtype C 4 photosynthesis and are important food and biomass crops, respectively. Both crops are grown in dense stands where shaded leaves can contribute a significant proportion of overall canopy productivity. This is because shaded leaves, despite intercepting little light, typically process light energy very efficiently for photosynthesis, when compared to light-saturated leaves at the top of the canopy. However, an apparently maladaptive loss in photosynthetic light-use efficiency as leaves become shaded has been shown to reduce productivity in these two species. It is unclear whether this is due to leaf aging or progressive shading from leaves forming above. This was resolved here by analysing photosynthesis in leaves of the same chronological age in the centre and exposed southern edge of field plots of these crops. Photosynthetic light-response curves were used to assess maximum quantum yield of photosynthesis; the key measure of photosynthetic capacity of a leaf in shade. Compared to the upper canopy, maximum quantum yield of photosynthesis of lower canopy leaves was significantly reduced in the plot centre; but increased slightly at the plot edge. This indicates loss of efficiency of shaded leaves is due not to aging, but to the altered light environment of the lower canopy, i.e., reduced light intensity and/or altered spectral composition. This work expands knowledge of the cause of this maladaptive shade response, which limits productivity of some of the world's most important crops.

Recent work has identified several proteins involved in starch granule initiation, the first step of starch synthesis. However, the degree of conservation in the granule initiation process remains poorly understood, especially among grass species differing in patterns of carbohydrate turnover in leaves, and granule morphology in the endosperm. We therefore compared mutant phenotypes of Hordeum vulgare (barley), Triticum turgidum (durum wheat), and Brachypodium distachyon defective in PROTEIN TARGETING TO STARCH 2 (PTST2), a key granule initiation protein. We report striking differences across species and organs. Loss of PTST2 from leaves resulted in fewer, larger starch granules per chloroplast and normal starch content in wheat, fewer granules per chloroplast and lower starch content in barley, and almost complete loss of starch in Brachypodium. The loss of starch in Brachypodium leaves was accompanied by high levels of ADP-glucose and detrimental effects on growth and physiology. Additionally, we found that loss of PTST2 increased granule initiations in Brachypodium amyloplasts, resulting in abnormal compound granule formation throughout the seed. These findings suggest that the importance of PTST2 varies greatly with genetic and developmental background and inform the extent to which the gene can be targeted to improve starch in crops.