David R. Causton

Regulation of assimilate partitioning in leaves

Concepts of the regulation of assimilate partitioning in leaves frequently consider only the allocation of carbon between sucrose and starch synthesis, storage and export. While carbohydrate metabolism accounts for a large proportion of assimilated carbon, such analyses provide only a restricted view of carbon metabolism and partitioning in leaf cells since photosynthetic carbon fixation provides precursors for all other biosynthetic pathways in the plant. Most of these precursors are required for biosynthesis of amino acids that form the building blocks for many compounds in plants. We have used leaf carbon : nitrogen ratios to calculate the allocation of photosynthetic electrons to the assimilation of nitrogen necessary for amino acid formation, and conclude that this allocation is variable but may be higher than values often quoted in the literature. Respiration is a significant fate of fixed carbon. In addition to supplying biosynthetic precursors, respiration is required for energy production and may also act, in both light and dark, to balance cellular energy budgets. We have used growth CO2 concentration and irradiance to modify source activity in Lolium temulentum in order to explore the interactions between photosynthetic carbon and nitrogen assimilation, assimilate production, respiration and export. It is demonstrated that there is a robust correlation between source activity and foliar respiration rates. Under some conditions concomitant increases in source activity and respiration may be necessary to support faster growth. In other conditions, increases in respiration appear to result from internal homeostatic mechanisms that may be candidate targets for increasing yield.

The application of MANOVA to analyse Arabidopsis thaliana metabolomic data from factorially designed experiments

Metabolomic technologies produce complex multivariate datasets and researchers are faced with the daunting task of extracting information from these data. Principal component analysis (PCA) has been widely applied in the field of metabolomics to reduce data dimensionality and for visualising trends within the complex data. Although PCA is very useful, it cannot handle multi-factorial experimental designs and, often, clear trends of biological interest are not observed when plotting various PC combinations. Even if patterns are observed, PCA provides no measure of their significance. Multivariate analysis of variance (MANOVA) applied to these PCs enables the statistical evaluation of main treatments and, more importantly, their interactions within the experimental design. The power and scope of MANOVA is demonstrated through two different factorially designed metabolomic investigations using Arabidopsis ethylene signalling mutants and their wild-type. One investigation has multiple experimental factors including challenge with the economically important pathogen Botrytis cinerea and also replicate experiments, while the second has different sample preparation methods and one level of replication ‘nested’ within the design. In both investigations there are specific factors of biological interest and there are also factors incorporated within the experimental design, which affect the data. The versatility of MANOVA is displayed by using data from two different metabolomic techniques; profiling using direct injection mass spectroscopy (DIMS) and fingerprinting using fourier transform infra-red (FT-IR) spectroscopy. MANOVA found significant main effects and interactions in both experiments, allowing a more complete and comprehensive interpretation of the variation within each investigation, than with PCA alone. Canonical variate analysis (CVA) was applied to investigate these effects and their biological significance. In conclusion, the application of MANOVA followed by CVA provided extra information than PCA alone and proved to be a valuable statistical addition in the overwhelming task of analysing metabolomic data.

Cellular basis of the effects of gibberellin and the pro gene on stem growth in tomato.

Tomato (Lycopersicon esculentum Mill.) plants homozygous for the mutant pro gene, exhibiting the distinctive procera phenotype, appeared virtually identical to gibberellic acid (GA3)-treated isogenic normal plants. The pro gene and GA3 caused analogous increases in internode length, and in the length and number of cells in the outer cell layers of each internode. Internode number was also increased by pro and GA3 over the period of the experiment. Despite their greater length, the internodes of GA3-treated and pro plants reached their final size within a time period similar to that of internodes of untreated normal plants. The pro mutant itself was responsive to GA3, especially in the seedling stage, but the proportional increase in height seen in the later stages of growth was less than that of normal plants.

UV responses of Lolium perenne raised along a latitudinal gradient across Europe: a filtration study

Lolium perenne (cv. AberDart) was grown at 14 locations along a latitudinal gradient across Europe (37-68°N) to study the impact of ultraviolet radiation (UV) and climate on aboveground growth and foliar UV-B absorbing compounds. At each location, plants were grown outdoors for 5 weeks in a replicated UV-B filtration experiment consisting of open, UV-B transparent (cellulose diacetate) and UV-B opaque (polyester) environments. Fourier transform-infrared spectroscopy was used to compare plant metabolite profiles in relation to treatment and location. UV radiation and climatic parameters were determined for each location from online sources and the data were assessed using a combination of anova and multiple regression analyses. Most of the variation in growth between the locations was attributable to the combination of climatic parameters, with minimum temperature identified as an important growth constraint. However, no single environmental parameter could consistently account for the variability in plant growth. Concentrations of foliar UV-B absorbing compounds showed a positive trend with solar UV across the latitudinal gradient; however, this relationship was not consistent in all treatments. The most striking experimental outcome from this study was the effect of presence or absence of filtration frames on UV-absorbing compounds. Overall, the study demonstrates the value of an European approach in studying the impacts of natural UV across a large latitudinal gradient. We have shown the feasibility of coordinated UV filtration at multiple sites but have also highlighted the need for open controls and careful interpretation of plant responses.

Acclimation of the summer annual species, Lolium temulentum, to CO2 enrichment

Abstract.Lolium temulentum L. Ba 3081 was grown hydroponically in air (350 μmol mol−1 CO2) and elevated CO2 (700 μmol mol−1 CO2) at two irradiances (150 and 500 μmol m−2 s−1) for 35 days at which point the plants were harvested. Elevated CO2 did not modify relative growth rate or biomass at either irradiance. Foliar carbon-to-nitrogen ratios were decreased at elevated CO2 and plants had a greater number of shorter tillers, particularly at the lower growth irradiance. Both light-limited and light-saturated rates of photosynthesis were stimulated. The amount of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco) protein was increased at elevated CO2, but maximum extractable Rubisco activities were not significantly increased. A pronounced decrease in the Rubisco activation state was found with CO2 enrichment, particularly at the higher growth irradiance. Elevated-CO2-induced changes in leaf carbohydrate composition were small in comparison to those caused by changes in irradiance. No CO2-dependent effects on fructan biosynthesis were observed. Leaf respiration rates were increased by 68% in plants grown with CO2 enrichment and low light. We conclude that high CO2 will only result in increased biomass if total light input favourably increases the photosynthesis-to-respiration ratio. At low irradiances, biomass is more limited by increased rates of respiration than by CO2-induced enhancement of photosynthesis.

Changes in percentage organic carbon content during ontogeny.

Changes in percentage organic carbon content were assessed during the first five weeks of growth of Uniculm barley (Hordeum vulgare) and Brussels sprouts (Brassica oleracea) plants grown in controlled-environment conditions at two constant temperatures, 16° and 22°C. Foliage (leaf laminae), stem, and root material was assayed in both species, together with leaf sheaths of barley and cotyledon laminae of Brussels sprouts. In barley, there was a decline in percentage organic carbon content with increasing foliage age in plants grown at 22°C, but in sheath material there was no significant change at either temperature. Root material showed a decline in percentage carbon content at both growth temperatures, whereas stems showed the opposite trend. Similar results were found in Brussels sprouts, with an overall decline in percentage carbon content in foliage at 22°C and a rise in stem material at both growth temperatures. However, roots showed no significant change in percentage carbon content over the experimental period. The results demonstrate that percentage organic carbon content may change during plant growth.

Acclimation of Lolium Temulentum to Growth at Elevated CO 2

Stimulation of photosynthesis and growth at elevated CO2 has been demonstrated in a variety of plant species, at least in the short-term (1,2,3). Long-term growth with CO2 enrichment frequently results in marked increases in foliar carbohydrate accumulation. Many studies on CO2 enrichment have examined species that accumulate sucrose and starch in their leaves. Relatively few have concentrated on plants that partition a large proportion of their assimilate into other sugars or sugar alcohols. We have therefore examined the responses of the fructan-forming monocotyledonous ryegrass Lolium temulentum to continuous growth in elevated CO2 (700 μmol mol−1) at two irradiances, 500 μmol m−2 s−1 (HL) and 150 μmol m−2 s−1 (LL).