Simple and robust determination of the activity signature of key carbohydrate metabolism enzymes for physiological phenotyping in model and crop plants.

The analysis of physiological parameters is important to understand the link between plant phenotypes and their genetic bases, and therefore is needed as an important element in the analysis of model and crop plants. The activities of enzymes involved in primary carbohydrate metabolism have been shown to be strongly associated with growth performance, crop yield, and quality, as well as stress responses. A simple, fast, and cost-effective method to determine activities for 13 key enzymes involved in carbohydrate metabolism has been established, mainly based on coupled spectrophotometric kinetic assays. The comparison of extraction buffers and requirement for dialysis of crude protein extracts resulted in a universal protein extraction protocol, suitable for the preparation of protein extracts from different organs of various species. Individual published kinetic activity assays were optimized and adapted for a semi-high-throughput 96-well assay format. These assays proved to be robust and are thus suitable for physiological phenotyping, enabling the characterization and diagnosis of the physiological state. The potential of the determination of distinct enzyme activity signatures as part of a physiological fingerprint was shown for various organs and tissues from three monocot and five dicot model and crop species, including two case studies with external stimuli. Differential and specific enzyme activity signatures are apparent during inflorescence development and upon in vitro cold treatment of young inflorescences in the monocot ryegrass, related to conditions for doubled haploid formation. Likewise, treatment of dicot spring oilseed rape with elevated CO2 concentration resulted in distinct patterns of enzyme activity responses in leaves.

Hormonal and metabolic regulation of source-sink relations under salinity and drought: from plant survival to crop yield stability.

Securing food production for the growing population will require closing the gap between potential crop productivity under optimal conditions and the yield captured by farmers under a changing environment, which is termed agronomical stability. Drought and salinity are major environmental factors contributing to the yield gap ultimately by inducing premature senescence in the photosynthetic source tissues of the plant and by reducing the number and growth of the harvestable sink organs by affecting the transport and use of assimilates between and within them. However, the changes in source-sink relations induced by stress also include adaptive changes in the reallocation of photoassimilates that influence crop productivity, ranging from plant survival to yield stability. While the massive utilization of -omic technologies in model plants is discovering hundreds of genes with potential impacts in alleviating short-term applied drought and salinity stress (usually measured as plant survival), only in relatively few cases has an effect on crop yield stability been proven. However, achieving the former does not necessarily imply the latter. Plant survival only requires water status conservation and delayed leaf senescence (thus maintaining source activity) that is usually accompanied by growth inhibition. However, yield stability will additionally require the maintenance or increase in sink activity in the reproductive structures, thus contributing to the transport of assimilates from the source leaves and to delayed stress-induced leaf senescence. This review emphasizes the role of several metabolic and hormonal factors influencing not only the source strength, but especially the sink activity and their inter-relations, and their potential to improve yield stability under drought and salinity stresses.