Analyzing anatomy over three dimensions unpacks the differences in mesophyll diffusive area between sun and shade Vitis vinifera leaves.

Leaves grown at different light intensities exhibit considerable differences in physiology, morphology and anatomy. Because plant leaves develop over three dimensions, analyses of the leaf structure should account for differences in lengths, surfaces, as well as volumes. In this manuscript, we set out to disentangle the mesophyll surface area available for diffusion per leaf area (S m,LA) into underlying one-, two- and three-dimensional components. This allowed us to estimate the contribution of each component to S m,LA, a whole-leaf trait known to link structure and function. We introduce the novel concept of a 'stomatal vaporshed,' i.e. the intercellular airspace unit most closely connected to a single stoma, and use it to describe the stomata-to-diffusive-surface pathway. To illustrate our new theoretical framework, we grew two cultivars of Vitis vinifera L. under high and low light, imaged 3D leaf anatomy using microcomputed tomography (microCT) and measured leaf gas exchange. Leaves grown under high light were less porous and thicker. Our analysis showed that these two traits and the lower S m per mesophyll cell volume (S m,Vcl) in sun leaves could almost completely explain the difference in S m,LA. Further, the studied cultivars exhibited different responses in carbon assimilation per photosynthesizing cell volume (A Vcl). While Cabernet Sauvignon maintained A Vcl constant between sun and shade leaves, it was lower in Blaufränkisch sun leaves. This difference may be related to genotype-specific strategies in building the stomata-to-diffusive-surface pathway.

Effects of Provenance, Growing Site, and Growth on Quercus robur Wood Anatomy and Density in a 12-Year-Old Provenance Trial.

Vessels are responsible for an efficient and safe water transport in angiosperm xylem. Whereas large vessels efficiently conduct the bulk of water, small vessels might be important under drought stress or after winter when large vessels are embolized. Wood anatomy can adjust to the environment by plastic adaptation, but is also modified by genetic selection, which can be driven by climate or other factors. To distinguish between plastic and genetic components on wood anatomy, we used a Quercus robur trial where trees from ten Central European provenances were planted in three locations in Austria along a rainfall gradient. Because wood anatomy also adjusts to tree size and in ring-porous species, the vessel size depends on the amount of latewood and thereby ring width, we included tree size and ring width in the analysis. We found that the trees' provenance had a significant effect on average vessel area (VA), theoretical specific hydraulic conductivity (Ks), and the vessel fraction (VF), but correlations with annual rainfall of provenances were at best weak. The trial site had a strong effect on growth (ring width, RW), which increased from the driest to the wettest site and wood density (WD), which increased from wet to dry sites. Significant site x provenance interactions were seen only for WD. Surprisingly, the drier site had higher VA, higher VF, and higher Ks. This, however, is mainly a result of greater RW and thus a greater proportion of latewood in the wetter forest. The average size of vessels > 70 µm diameter increased with rainfall. We argue that Ks, which is measured per cross-sectional area, is not an ideal parameter to compare the capacity of ring-porous trees to supply leaves with water. Small vessels (<70 µm) on average contributed only 1.4% to Ks, and we found no evidence that their number or size was adaptive to aridity. RW and tree size had strong effect on all vessel parameters, likely via the greater proportion of latewood in wide rings. This should be accounted for when searching for wood anatomical adaptations to the environment.

Container volume affects drought experiments in grapevines: Insights on xylem anatomy and time of dehydration.

Plant stress experiments are commonly performed with plants grown in containers to better control environmental conditions. Nevertheless, the container can constrain plant growth and development, and this confounding effect is generally ignored, particularly in studies on woody species. Here, we evaluate the effect of the container volume in drought experiments using grapevine as a model plant. Grapevines grown in small (7 L, S) or large (20 L, L) containers were subjected to drought stress and rewatering treatments. We monitored plant stomatal conductance (gs ), midday stem water potential (Ψs ), and photosynthetic rate (AN ) throughout the experiment. The effect of the container volume on the stem and petiole xylem anatomy, as well as on the total leaf area (LA), was assessed before drought imposition. The results showed that LA did not differ between plants in L or S containers, but S vines exhibited a higher theoretical hydraulic conductance at the petiole level. Under drought L and S similarly reduced gs and AN , but plants in S containers reached lower Ψs than those in L. Nevertheless, upon rewatering droughted plants in S containers exhibited a faster stomata re-opening than those in L, probably as a consequence of the differences in the stress degree experienced and the biochemical adjustment at the leaf level. Therefore, a suitable experimental design should consider the container volume used in relation to the desired traits to be studied for unbiased results.

Assessing adaptive and plastic responses in growth and functional traits in a 10-year-old common garden experiment with pedunculate oak (Quercus robur L.) suggests that directional selection can drive climatic adaptation.

Understanding how tree species will respond to a future climate requires reliable and quantitative estimates of intra-specific variation under current climate conditions. We studied three 10-year-old common garden experiments established across a rainfall and drought gradient planted with nearly 10,000 pedunculate oak (Quercus robur L.) trees from ten provenances with known family structure. We aimed at disentangling adaptive and plastic responses for growth (height and diameter at breast height) as well as for leaf and wood functional traits related to adaptation to dry environments. We used restricted maximum likelihood approaches to assess additive genetic variation expressed as narrow-sense heritability (h2), quantitative trait differentiation among provenances (QST), and genotype-by-environment interactions (GxE). We found strong and significant patterns of local adaptation in growth in all three common gardens, suggesting that transfer of seed material should not exceed a climatic distance of approximately 1°C under current climatic conditions, while transfer along precipitation gradients seems to be less stringent. Moreover, heritability reached 0.64 for tree height and 0.67 for dbh at the dry margin of the testing spectrum, suggesting significant additive genetic variation of potential use for future selection and tree breeding. GxE interactions in growth were significant and explained less phenotypic variation than origin of seed source (4% versus 10%). Functional trait variation among provenances was partly related to drought regimes at provenances origins but had moderate explanatory power for growth. We conclude that directional selection, either naturally or through breeding, is the most likely and feasible outcome for pedunculate oak to adapt to warmer and drier climate conditions in the future.

Difference in reproductive mode rather than ploidy explains niche differentiation in sympatric sexual and apomictic populations of Potentilla puberula.

Apomicts tend to have larger geographical distributional ranges and to occur in ecologically more extreme environments than their sexual progenitors. However, the expression of apomixis is typically linked to polyploidy. Thus, it is a priori not clear whether intrinsic effects related to the change in the reproductive mode or rather in the ploidy drive ecological differentiation. We used sympatric sexual and apomictic populations of Potentilla puberula to test for ecological differentiation. To distinguish the effects of reproductive mode and ploidy on the ecology of cytotypes, we compared the niches (a) of sexuals (tetraploids) and autopolyploid apomicts (penta-, hepta-, and octoploids) and (b) of the three apomictic cytotypes. We based comparisons on a ploidy screen of 238 populations along a latitudinal transect through the Eastern European Alps and associated bioclimatic, and soil and topographic data. Sexual tetraploids preferred primary habitats at drier, steeper, more south-oriented slopes, while apomicts mostly occurred in human-made habitats with higher water availability. Contrariwise, we found no or only marginal ecological differentiation among the apomictic higher ploids. Based on the pronounced ecological differences found between sexuals and apomicts, in addition to the lack of niche differentiation among cytotypes of the same reproductive mode, we conclude that reproductive mode rather than ploidy is the main driver of the observed differences. Moreover, we compared our system with others from the literature, to stress the importance of identifying alternative confounding effects (such as hybrid origin). Finally, we underline the relevance of studying ecological parthenogenesis in sympatry, to minimize the effects of differential migration abilities.

Asymmetric reproductive interference: The consequences of cross-pollination on reproductive success in sexual-apomictic populations of Potentilla puberula (Rosaceae).

Apomixis evolves from a sexual background and usually is linked to polyploidization. Pseudogamous gametophytic apomicts, which require a fertilization to initiate seed development, of various ploidy levels frequently co-occur with their lower-ploid sexual ancestors, but the stability of such mixed populations is affected by reproductive interferences mediated by cross-pollination. Thereby, reproductive success of crosses depends on the difference in ploidy levels of mating partners, that is, on tolerance of deviation from the balanced ratio of maternal versus paternal genomes. Quality of pollen can further affect reproductive success in intercytotype pollinations. Cross-fertilization, however, can be avoided by selfing which may be induced upon pollination with mixtures of self- and cross-pollen (i.e., mentor effects). We tested for reproductive compatibility of naturally co-occurring tetraploid sexuals and penta- to octoploid apomicts in the rosaceous species Potentilla puberula by means of controlled crosses. We estimated the role of selfing as a crossing barrier and effects of self- and cross-pollen quality as well as maternal: paternal genomic ratios in the endosperm on reproductive success. Cross-fertilization of sexuals by apomicts was not blocked by selfing, and seed set was reduced in hetero- compared to homoploid crosses. Thereby, seed set was negatively related to deviations from balanced parental genomic ratios in the endosperm. In contrast, seed set in the apomictic cytotypes was not reduced in hetero- compared to homoploid crosses. Thus, apomictic cytotypes either avoided intercytotype cross-fertilization through selfing, tolerated intercytotype cross-fertilizations without negative effects on reproductive success, or even benefitted from higher pollen quality in intercytotype pollinations. Our experiment provides evidence for asymmetric reproductive interference, in favor of the apomicts, with significantly reduced seed set of sexuals in cytologically mixed populations, whereas seed set in apomicts was not affected. Incompleteness of crossing barriers further indicated at least partial losses of a parental genomic endosperm balance requirement.