The EU and the Catalan Crisis

List of all differentially expressed proteins (n = 45) identified in the study. (XLSX 11 kb

Proteomics Analysis of Alfalfa Response to Heat Stress

<div><p>The proteome responses to heat stress have not been well understood. In this study, alfalfa (<i>Medicago sativa</i> L. cv. Huaiyin) seedlings were exposed to 25°C (control) and 40°C (heat stress) in growth chambers, and leaves were collected at 24, 48 and 72 h after treatment, respectively. The morphological, physiological and proteomic processes were negatively affected under heat stress. Proteins were extracted and separated by two-dimensional polyacrylamide gel electrophoresis (2-DE), and differentially expressed protein spots were identified by mass spectrometry (MS). Totally, 81 differentially expressed proteins were identified successfully by MALDI-TOF/TOF. These proteins were categorized into nine classes: including metabolism, energy, protein synthesis, protein destination/storage, transporters, intracellular traffic, cell structure, signal transduction and disease/defence. Five proteins were further analyzed for mRNA levels. The results of the proteomics analyses provide a better understanding of the molecular basis of heat-stress responses in alfalfa.</p></div

2-DE analysis of alfalfa leaf proteins extracted from alfalfa leaves grown under control(A), heat stress (40°C for 24(B), 48(C) and 72 h(D)) samples.

<p>A total of 600 µg proteins were separated by 2-DE as described in the material and methods and then visualized using CBB stain. Arrows indicate proteins that were 1.5 fold changes in respond to heat stress.</p

The functional category distribution of all identified proteins in response to heat stress.

<p>The functional category distribution of all identified proteins in response to heat stress.</p

Phenotypic and physiological changes in alfalfa seedlings under control and heat stress (40°C for 24, 48 and 72 h).

<p>(A). Phenotypic changes (B). RWC (C). Proline content. Three independent biological replicates were used in each treatment. Asterisks indicate statistically significant differences at <i>P</i><0.05. Bars represent standard errors of triplicate experiments.</p

Contrasting Effects of Long-Term Grazing and Clipping on Plant Morphological Plasticity: Evidence from a Rhizomatous Grass

<div><p>Understanding the mechanism of plant morphological plasticity in response to grazing and clipping of semiarid grassland can provide insight into the process of disturbance-induced decline in grassland productivity. In recent studies there has been controversy regarding two hypotheses: 1) grazing avoidance; and 2) growth limiting mechanisms of morphological plasticity in response to defoliation. However, the experimental evidence presented for the memory response to grazing and clipping of plants has been poorly reported. This paper reports on two experiments that tested these hypotheses in field and in a controlled environment, respectively. We examined the effects of long-term clipping and grazing on the functional traits and their plasticity for <i>Leymus chinensis</i> (Trin.) Tzvelev (the dominate species) in the typical-steppe grassland of Inner Mongolia, China. There were four main findings from these experiments. (i) The majority of phenotypic traits of <i>L</i>. <i>chinensis</i> tended to significantly miniaturize in response to long-term field clipping and grazing. (ii) The significant response of morphological plasticity with and without grazing was maintained in a hydroponic experiment designed to remove environmental variability, but there was no significant difference in <i>L</i>. <i>chinensis</i> individual size traits for the clipping comparison. (iii) Plasticity indexes of <i>L</i>. <i>chinensis</i> traits in a controlled environment were significantly lower than under field conditions indicating that plants had partial and slight memory effect to long-term grazing. (iv) The allometry of various phenotypic traits, indicated significant trade-offs between leaf and stem allocation with variations in plant size induced by defoliation, which were maintained only under grazing in the hydroponic controlled environment experiment. Taken together, our findings suggest that the morphological plasticity of <i>L</i>. <i>chinensis</i> induced by artificial clipping was different with that by livestock grazing. The miniaturization of plant size in long-term grazed grassland may reflect retained characteristics of dwarf memory for adaptation to long-term grazing by large herbivores.</p></div

Relationships between the difference-value of plasticity index (<i>PI</i>) of grazing and clipping treatments.

<p>Difference-value of <i>PI</i>: <i>PI</i> (<i>in situ</i>)–<i>PI</i> (hydroponics). The correlation between the differences in <i>PI</i> was tested by Pearson’s method (<i>r</i> = 0.87, <i>P</i> < 0.01). The gray area represents traits that exhibit a larger change in <i>PI</i> after clipping treatments than grazing treatments. Solid line: linear fit; dashed line: 1:1 line.</p

Effects of long-term grazing and clipping on phenotypic traits of <i>Leymus chinensis</i> (Trin.) Tzvelev individuals in field experiments.

<p>Symbols</p><p>(-), treatments that had negative effects on phenotypic traits</p><p>(+), treatments that had positive effects on phenotypic traits</p><p>(0), treatments that had no effects on phenotypic traits.</p><p>Other abbreviations are the same as those in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0141055#pone.0141055.t001" target="_blank">Table 1</a>.</p><p>Effects of long-term grazing and clipping on phenotypic traits of <i>Leymus chinensis</i> (Trin.) Tzvelev individuals in field experiments.</p

Differences in the plasticity index of <i>Leymus chinensis</i> (Trin.) Tzvelev phenotypic traits between field grazing and hydroponic conditions.

<p>Symbols: **, <i>P</i> < 0.01; *, <i>P</i> < 0.05; NS, <i>P</i> > 0.05.</p

Standardized major axis (<i>SMA</i>) regression slopes and their confidence intervals for the log-log transformed relationship between stem length and leaf length of <i>Leymus chinensis</i> (Trin.) Tzvelev in grazing exclusion and grazing groups.

<p>Symbols and abbreviations are as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0141055#pone.0141055.t001" target="_blank">Table 1</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0141055#pone.0141055.g001" target="_blank">Fig 1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0141055#pone.0141055.g004" target="_blank">Fig 4</a>.</p