System-level effects of CO2 and RuBisCO concentration on carbon isotope fractionation

ABSTRACTCarbon isotope biosignatures preserved in the Precambrian geologic record are primarily interpreted to reflect ancient cyanobacterial carbon fixation catalyzed by Form I RuBisCO enzymes. The average range of isotopic biosignatures generally follows that produced by extant cyanobacteria. However, this observation is difficult to reconcile with several environmental (e.g., temperature, pH, and CO2 concentrations), molecular and physiological factors that likely would have differed during the Precambrian and can produce fractionation variability in contemporary organisms that meets or exceeds that observed in the geologic record. To test a range of genetic and environmental factors that may have impacted ancient carbon isotope biosignatures, we engineered a mutant strain of the model cyanobacterium Synechococcus elongatus PCC 7942 that overexpresses RuBisCO and characterized the resultant physiological and isotope fractionation effects. We specifically investigated how both increased atmospheric CO2 concentrations and RuBisCO regulation influence cell growth, oxygen evolution rate, and carbon isotope fractionation in cyanobacteria. We found that >2% CO2 increases the growth rate of wild-type and mutant strains, and that the pool of active RuBisCO enzyme increases with increased expression. At elevated CO2, carbon isotope discrimination (εp) is increased by ~8‰, whereas RuBisCO overexpression does not significantly affect isotopic discrimination at all tested CO2 concentrations. Our results show that understanding the environmental factors that impact RuBisCO regulation, physiology, and evolution is crucial for reconciling microbially driven carbon isotope fractionation with the geologic record of organic and inorganic carbon isotope signatures.IMPORTANCECarbon isotope biosignatures preserved in the geologic record are interpreted to reflect the long-term evolution of microbial carbon fixation and provide the earliest evidence of life on Earth. RuBisCO enzymes, distinctive and early-evolved catalysts that fix atmospheric CO2, have likely been responsible for the bulk of primary productivity through Earth history. Thus, a comprehensive understanding of the molecular, physiological, environmental, and evolutionary factors that influence the isotopic discrimination of cyanobacteria that utilize RuBisCO is essential for the interpretation of ancient isotopic biosignatures. For example, the vastly different atmospheric CO2 levels that characterized the Precambrian may have influenced the expression and regulation of the ancient RuBisCO protein complex. These observations underscore the need to consider how a broader range of environmental conditions and subcellular processes may have shaped isotopic discrimination over geologic time. In this study, we establish a cyanobacterial metabolic-engineering strategy that can test such hypotheses and offer insights into the biogeochemical record of life.

Integration of Physiological and Molecular Traits Would Help to Improve the Insights of Drought Resistance in Highbush Blueberry Cultivars

Water deficit or drought is one of the most severe factors limiting plant yield or fruit quality. Thus, water availability for irrigation is decisive for crop success, such as the case of highbush blueberry (Vaccinium corymbosum L.). Therefore, drought stress may compromise blueberry production due to lower fruit weight or fruit yield. Despite this, it is unclear if there is any difference in the response of blueberry cultivars to water deficit, either in terms of physiological and molecular parameters, or in terms of their sensitivity or resistance to drought. In this study, we determined the effect of drought on different physiological parameters in blueberry plants (relative water content (RWC), photochemical efficiency of photosystem II (Fv/Fm), Carbon Isotopic Discrimination, and proline content) in six V. corymbosum cultivars. We also explored molecular responses in terms of gene expression coding for late embryogenesis abundant proteins. Finally, we estimated cultivar water deficit resistance using an integrative model based on physiological results. Upon water deficit conditions, we found reductions in Fv/Fm, RWC, and isotopic discrimination of 13C (Δ13C), while proline content increased significantly for all cultivars. Additionally, we also found differences in the estimated water deficit resistance index. These results indicate differences in water deficit resistance, possibly due to variations in cultivars’ genetic composition.

A Comparison of the Variable J and Carbon-Isotopic Composition of Sugars Methods to Assess Mesophyll Conductance from the Leaf to the Canopy Scale in Drought-Stressed Cherry

Conductance of CO2 across the mesophyll (Gm) frequently constrains photosynthesis (PN) but cannot be measured directly. We examined Gm of cherry (Prunus avium L.) subjected to severe drought using the variable J method and carbon-isotopic composition (δ13C) of sugars from the centre of the leaf, the leaf petiole sap, and sap from the largest branch. Depending upon the location of the plant from which sugars are sampled, Gm may be estimated over scales ranging from a portion of the leaf to a canopy of leaves. Both the variable J and δ13C of sugars methods showed a reduction in Gm as soil water availability declined. The δ13C of sugars further from the source of their synthesis within the leaf did not correspond as closely to the diffusive and C-isotopic discrimination conditions reflected in the instantaneous measurement of gas exchange and chlorophyll-fluorescence utilised by the variable J approach. Post-photosynthetic fractionation processes and/or the release of sugars from stored carbohydrates (previously fixed under different environmental and C-isotopic discrimination conditions) may reduce the efficacy of the δ13C of sugars from leaf petiole and branch sap in estimating Gm in a short-term study. Consideration should be given to the spatial and temporal scales at which Gm is under observation in any experimental analysis.