Diversity and functional differentiation of renewal buds in temperate herbaceous plants.

Spring regrowth in temperate perennials relies on renewal buds, which form a key component in the shoot growth cycle. Still, we possess almost no information on these renewal buds, which is becoming more pressing with the current climate change. Most existing studies concentrated on easy-to-study aboveground buds of woody plants, whose morphology has largely been linked to frost protection. It is not clear to what extent these findings apply also to herbaceous species. We therefore examined protective traits and preformation of winter renewal buds in 379 species of temperate herbs, and tested how these traits are distributed across the phylogeny and related to other bud bank and whole-plant traits. We identified a major gradient from few, large, highly preformed, scale-covered buds associated with larger belowground storage organs deep in the soil, to small, numerous, less preformed, and naked buds near the soil surface. Belowground renewal buds of temperate herbs show several distinct strategies for winter survival and spring regrowth that might affect their response to changing winter and early spring conditions. Renewal bud traits are driven not only by frost protection but also by protection of the apical meristem from mechanical disturbance in the soil.

Species phylogeny, ecology, and root traits as predictors of root exudate composition.

Root traits including root exudates are key factors affecting plant interactions with soil and thus play an important role in determining ecosystem processes. The drivers of their variation, however, remain poorly understood. We determined the relative importance of phylogeny and species ecology in determining root traits and analyzed the extent to which root exudate composition can be predicted by other root traits. We measured different root morphological and biochemical traits (including exudate profiles) of 65 plant species grown in a controlled system. We tested phylogenetic conservatism in traits and disentangled the individual and overlapping effects of phylogeny and species ecology on traits. We also predicted root exudate composition using other root traits. Phylogenetic signal differed greatly among root traits, with the strongest signal in phenol content in plant tissues. Interspecific variation in root traits was partly explained by species ecology, but phylogeny was more important in most cases. Species exudate composition could be partly predicted by specific root length, root dry matter content, root biomass, and root diameter, but a large part of variation remained unexplained. In conclusion, root exudation cannot be easily predicted based on other root traits and more comparative data on root exudation are needed to understand their diversity.

Inflorescence preformation prior to winter: a surprisingly widespread strategy that drives phenology of temperate perennial herbs.

Organ preformation in overwintering buds of perennial plants has been known for almost two centuries. It is hypothesized to underlie fast growth and early flowering, but its frequency, phylogenetic distribution, and ecological relevance have never been systematically examined. We microscopically observed inflorescence preformation in overwintering buds (IPB) in the autumn. We studied a phylogenetically and ecologically representative set of 330 species of temperate perennial angiosperms and linked these observations with quantitative data on species' flowering phenology, genome size, and ecology. IPB was observed in 34% of species examined (in 14% species the stamens and/or pistils were already developed). IPB is fairly phylogenetically conserved and frequent in many genera (Alchemilla, Carex, Euphorbia, Geranium, Primula, Pulmonaria) or families (Ranunculaceae, Euphorbiaceae, Violaceae, Boraginaceae). It was found in species of any genome size, although it was almost universal in those with large genomes. Compared with non-IPB species, IPB species flowered 38 d earlier on average and were more common in shaded and undisturbed habitats. IPB is a surprisingly widespread adaptation for early growth in predictable (undisturbed) conditions. It contributes to temporal niche differentiation and has important consequences for understanding plant phenology, genome size evolution, and phylogenetic structure of plant communities.

Disentangling phylogenetic and functional components of shape variation among shoot apical meristems of a wide range of herbaceous angiosperms.

PREMISE: The shoot apical meristem (SAM) is the basic determinant of plant body organization, but interspecific variation in SAM shape and its relationship to stem and leaf morphological traits is not well known. Here we tested the hypothesis that different SAM shapes are associated with specific shoot traits of the plant body and examined the phylogenetic conservatism of these relationships. METHODS: We used geometric morphometrics of SAM outlines for a phylogenetically representative set of 110 herbaceous angiosperms and examined their relationship to a number of shoot traits. RESULTS: We found large variations in SAM shapes across angiosperm lineages, but covering only a subset of geometrically possible shapes. Part of this variation was allometric (due to SAM size), but the dominant shape variation (dome-shaped vs. flat surface) was size-independent and strongly phylogenetically conserved. SAM shapes were largely independent of their cell size and therefore of the number of cells involved. Different patterns in shape variation of outer and inner SAM boundaries were associated with stem thickness, leaf area, and leafiness of the stem. CONCLUSIONS: The findings show that geometric interdependence of meristem zones gives rise to correlations among organ numbers, sizes, and their proportions. Phylogenetic conservatism in these correlations indicates conservatism in regulatory processes that underlie the correlations, or the individual traits, that give rise to plant architecture.

Shoot apical meristem and plant body organization: a cross-species comparative study.

Background and Aims: The shoot apical meristem (SAM) is the key organizing element in the plant body and is responsible for the core of plant body organization and shape. Surprisingly, there are almost no comparative data that would show links between parameters of the SAM and whole-plant traits as drivers of the plant's response to the environment. Methods: Interspecific differences in SAM anatomy were examined in 104 perennial herbaceous angiosperms. Key Results: There were differences in SAM parameters among individual species, their phylogenetic patterns, and how their variation is linked to variation in plant above-ground organs and hence species' environmental niches. SAM parameters were correlated with the size-related traits of leaf area, seed mass and stem diameter. Of the two key SAM parameters (cell size and number), variation in all organ traits was linked more strongly to cell number, with cell size being important only for seed mass. Some of these correlations were due to shared phylogenetic history (e.g. SAM diameter versus stem diameter), whereas others were due to parallel evolution (e.g. SAM cell size and seed mass). Conclusion: These findings show that SAM parameters provide a functional link among sizes and numbers of plant organs, constituting species' environmental responses.

Plant Origin, but Not Phylogeny, Drive Species Ecophysiological Response to Projected Climate.

Knowledge of the relationship between environmental conditions and species traits is an important prerequisite for understanding determinants of community composition and predicting species response to novel climatic conditions. Despite increasing number of studies on this topic, our knowledge on importance of genetic differentiation, plasticity and their interactions along larger sets of species is still limited especially for traits related to plant ecophysiology. We studied variation in traits related to growth, leaf chemistry, contents of photosynthetic pigments and activity of antioxidative enzymes, stomata morphology and photosynthetic activity across eight Impatiens species growing along altitudinal gradients in Himalayas cultivated in three different temperature regimes and explored effects of among species phylogenetic relationships on the results. Original and target climatic conditions determine trait values in our system. The traits are either highly plastic (e.g., APX, CAT, plant size, neoxanthin, ß-carotene, chlorophyll a/b, DEPSC) or are highly differentiated among populations (stomata density, lutein production). Many traits show strong among population differentiation in degree of plasticity and direction in response to environmental changes. Most traits indicate that the species will profit from the expected warming. This suggests that different processes determine the values of the different traits and separating the importance of genetic differentiation and plasticity is crucial for our ability to predict species response to future climate changes. The results also indicate that evolution of the traits is not phylogenetically constrained but including phylogenetic information into the analysis may improve our understanding of the trait-environment relationships as was apparent from the analysis of SLA.

Transgenic ipt tobacco overproducing cytokinins overaccumulates phenolic compounds during in vitro growth.

We present evidence that overproduction of endogenous cytokinins (CK) caused stress response in non-rooting Pssu-ipt transgenic tobacco (Nicotiana tabacum L.) grown in vitro. It was demonstrated by overaccumulation of phenolic compounds, synthesis of pathogenesis related proteins (PR proteins), and increase in peroxidase (POD) activities. Immunolocalization of zeatin and also PR-1b protein on leaf cryo-sections proved their accumulation in all mesophyll cells of transgenic tobacco contrary to control non-transgenic plants. Intensive blue autofluorescence of phenolic compounds induced by UV in cross-sections of leaf midrib showed enhanced contents of phenolics in transgenic tobacco compared with controls, nevertheless, no significant difference between both plant types was found in leaf total lignin content. Transgenic plantlets exhibited higher peroxidase activities of both soluble and ionically bound fractions compared with controls. HPLC analysis of phenolic acids confirmed the increase in all phenolic acids in transgenic tobacco except for salicylic acid (SA). The effect of high phenolic content on rooting of transgenic tobacco is discussed.

Tobacco susceptibility to Potato virus Y(NTN) infection is affected by grafting and endogenous cytokinin content.

Faster or stronger response to pathogen occurs if plants undergo prior priming. Cytokinins seem to be also involved in plant priming and in response to pathogens. Susceptibility to Potato virus Y(NTN) (PVY(NTN)) was studied in transgenic cytokinin overproducing (Pssu-ipt) tobacco and compared with nontransgenic plants. Since cytokinin overproduction inhibits development of plant roots and grafting overcomes this limitation, both types were grown as rooted and/or grafted plants to check also the effect of grafting. Control rooted tobacco (C), the most susceptible to PVY(NTN), showed always symptoms during the infection together with the rising virus content and a systemic response, such as accumulation of H2O2, salicylic acid (SA) and other phenolic acids, and stress-induced enzyme activities. In transgenic and grafted plants, the response to PVY(NTN) was dependent on protective mechanisms activated prior to the inoculation. In Pssu-ipt tobacco, cytokinin active forms and SA contents exceeded manifold their content in C. Grafting promoted the accumulation of phenolics, but SA, and stimulated peroxidase activities. Thus, the pre-infection barrier established in both transgenic and grafted plants helped to suppress partly the virus multiplication and resulted in milder symptom development. However, only the synergic effect of both grafting and the high cytokinins led to PVY(NTN) tolerance in transgenic grafts. Possible mechanisms were discussed.

Three-dimensional reconstruction of anomalous chloroplasts in transgenic ipt tobacco.

Anomalies in the ultrastructure of chloroplasts, from transgenic ipt tobacco, overproducing endogenous cytokinins (CKs) were studied. Detailed analyses of CKs and their metabolites showed that Pssu-ipt tobacco contained enhanced contents of CKs both in leaves and in isolated chloroplasts. The role of CKs in the formation of anomalous structures is suggested. Pssu-ipt chloroplasts frequently formed the distinct peripheral reticulum with a system of caverns that often involved mitochondria and/or peroxisomes. Large crystalloids, which were found in chloroplasts of Pssu-ipt, occupied up to 16% of chloroplast volume. We suggested that the crystalloids were formed by LHC II aggregates. This was supported by analysis of the fluorescence emission spectra at 77 degrees K, chlorophyll a/b ratio, immunogold staining of the structures, and crystallographic unit size analysis.