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Item Functional dissection of ERD14 phosphorylation-dependent calcium binding activity(2014-12-11) Chacha, Allen R.; Randall, Stephen Karl, 1953-; Watson, John; Kusmierczyk, AndrewDrought and cold conditions are among the major factors affecting plant growth and crop production globally. Dehydrins are group II late embryogenesis abundant (LEA) proteins characterized by a conserved K-region (EKKGIMDKIKEKLPG consensus sequence) that accumulate in many plants during drought, low temperature, and high salinity to confer stress tolerance. While it has been demonstrated that overexpression of dehydrins improves cold tolerance in various crop plants, the mechanism leading to cold tolerance is still unclear. Previous studies reported phosphorylation of AtERD14 dehydrin by casein kinase II (CKII) led to an increase in calcium binding activity. Mass spectroscopy analysis determined that the phosphorylation was localized to a poly-serine (S) region. To further characterize the S-region, GST fused ERD14 mutants were created via site-directed mutagenesis and deletion of either the amino or carboxyl ends of ERD14 via the QuickChange® Multi Site-Directed Mutagenesis Kit. Phosphorylation of purified mutant proteins by CKII was analyzed via gel shift and direct phosphorylation assays. The effect of phosphorylation on calcium binding activity was also analyzed. Results showed the serine (S) residue at position 83 was crucial to phosphorylation-dependent molecular mass shift and Ca2+-binding activities followed by the serine residue at position 85 in importance. Mutation of serines at positions 83, 84, and 85 completely eliminated the phosphorylation-dependent gel shift and calcium binding. Examination of truncation mutants determined the N-terminal was an important region for protein structure modification and phosphorylation ability leading to Ca2+ activation. Calcium binding activity of the truncated mutants indicated the calcium binding site was localized in the region between the S-region and the K-region near the C-terminal end. To characterize the acidic dehydrins contribution to cold tolerance in vivo, three single (erd10, erd14, cor47) knockouts (KOs) were characterized. Single KOs produced no cold sensitive phenotype indicating the need for multiple dehydrin KOs in Arabidopsis in order to potentially produce a cold sensitive phenotype.Item Observed increasing water constraint on vegetation growth over the last three decades(Springer Nature, 2021-06-18) Jiao, Wenzhe; Wang, Lixin; Smith, William K.; Chang, Qing; Wang, Honglang; D’Odorico, Paolo; Earth and Environmental Sciences, School of ScienceDespite the growing interest in predicting global and regional trends in vegetation productivity in response to a changing climate, changes in water constraint on vegetation productivity (i.e., water limitations on vegetation growth) remain poorly understood. Here we conduct a comprehensive evaluation of changes in water constraint on vegetation growth in the extratropical Northern Hemisphere between 1982 and 2015. We document a significant increase in vegetation water constraint over this period. Remarkably divergent trends were found with vegetation water deficit areas significantly expanding, and water surplus areas significantly shrinking. The increase in water constraints associated with water deficit was also consistent with a decreasing response time to water scarcity, suggesting a stronger susceptibility of vegetation to drought. We also observed shortened water surplus period for water surplus areas, suggesting a shortened exposure to water surplus associated with humid conditions. These observed changes were found to be attributable to trends in temperature, solar radiation, precipitation, and atmospheric CO2. Our findings highlight the need for a more explicit consideration of the influence of water constraints on regional and global vegetation under a warming climate.