An innovative light chamber for measuring photosynthesis by three-dimensional plant organs.

BACKGROUND: In plants, three-dimensional (3-D) organs such as inflorescences or fruits carry out photosynthesis and thus play a significant role in carbon assimilation and yield. However, this contribution has been poorly characterized because there is no reliable method for measuring photosynthesis by 3-D organs. One of the major challenges is ensuring the uniform irradiation of samples that are placed within a sealed chamber. RESULTS: In this study, we developed an innovative chamber with homogeneous lighting that can be used to measure photosynthesis by large 3-D organs. It consisted of a 15-cm-long sealed transparent cylinder that was surrounded by a decagonal prismatic light source, made up of a mixture of red and blue LEDs. We characterized irradiance homogeneity within the chamber at a resolution level of 1 cm and 10°. Photosynthetic photon flux density (PPFD) along the central axis of the chamber could be set to any value between 100 and 1100 µmol m-2 s-1. The coefficient of variation for the irradiation values found throughout the chamber was 10% and that for the ratio of red-to-blue spectra was less than 1.5%. The temperature of the sample was regulated to stay within 1 °C of the target temperature, regardless of PPFD. We compared the performance of our device with that of a commercially available device employing unidirectional lighting. Specifically, we examined net photosynthesis in two sample types-wheat ears and grape clusters-at varying PPFD levels. CONCLUSIONS: The devices gave similar estimates of dark respiration, regardless of sample type or age. Conversely, net photosynthesis started to become asymptotic at lower irradiance levels in our device than in the conventional device because apparent quantum yield was three times higher. When examining the effects of irradiance heterogeneity, it was clear that biased estimates could result from systems employing unidirectional light sources. Our results also confirmed that our chamber could be a useful tool for obtaining more accurate estimates of photosynthesis by 3-D organs.

Source-sink partitioning. Do we need Münch?

The simulation of phloem translocation by the Münch theory commonly uses resistances from sources to sinks: the resistances are therefore regarded as important in partitioning. Although resistance is generally a set constant, it is in fact strongly affected by viscosity, and thus the concentration of the transported solute. In this paper, the model of partitioning proposed by Minchin et al. was first corrected for variations in viscosity. The model was further modified, with the source considered as an activity of solute production rather than as a compartment concentration. When so defined, the source cannot differ from the sum of sink activities, largely outdating the source- or sink-limitation concepts. The corrected model confined the effect of resistances on the partitioning to low source activities. In the example of wheat grain filling analysed, such activities would be so low that they would correspond only to pathological conditions. In that case, the use of resistances in modelling is therefore just a mathematical burden, not even easily quantifiable since they are related to anatomical traits that are difficult to access. Leaving out resistances, it becomes easy to calculate the sink activities directly from the source activities, using an intuitive, accessible parametrization. The conditions for such a simplification are discussed.

Sucrose:Fructan 6-Fructosyltransferase, a Key Enzyme for Diverting Carbon from Sucrose to Fructan in Barley Leaves.

Sucrose:sucrose 6-fructosyltransferase, an enzyme activity recently identified in fructan-accumulating barley (Hordeum vulgare) leaves, was further characterized. The purified enzyme catalyzed the transfer of a fructosyl group from sucrose to various acceptors. It displayed some [beta]-fructosidase (invertase) activity, indicating that water could act as fructosyl acceptor. Moreover, it transferred the fructosyl residue of unlabeled sucrose to [U-14C]Glc, producing [U-14C]sucrose and unlabeled glucose. Most significantly for fructan synthesis, the enzyme used as acceptors but not as donors a variety of oligofructans containing [beta](2->1)- and [beta](2->6)-linked fructosyl moieties. Thus, it acted as a general sucrose:fructan fructosyltransferase. The products formed by the enzyme from sucrose and various purified, structurally characterized oligofructans were analyzed by liquid chromatography and identified by comparison with structurally characterized standards. The results showed that the enzyme formed exclusively [beta](2->6) fructosyl-fructose linkages, either initiating or elongating a fructan chain of the phlein type. We propose, therefore, to rename the purified enzyme sucrose:fructan 6-fructosyltransferase.

Fructan chemical structure and sensitivity to an exohydrolase.

A fructan exohydrolase selective for (2----1)-linked terminal fructosyl linkages, isolated from barley (Hordeum vulgare L. cv. Morex) stems and leaf sheaths, was used to elucidate the chemical structures of several oligomeric fructans extracted from liliaceous and graminaceous species. Products released by enzymic and mild acid hydrolysis were separated by reversed-phase high-performance liquid chromatography. Gas-liquid chromatography-mass spectrometry of partially methylated alditol acetates permitted unequivocal deduction of many linkage sequences, first of the hydrolysis products and then of the original oligomers. We found that bifurcose, a tetrasaccharide formed by addition of a fructosyl unit to O-6 of the central fructose residue of 1-kestose, was a central molecule in the generation of the branched, oligomeric fructans of wheat (Triticum aestivum L. cv. Fidel). These arise by the extension of both (2----1)- and (2----6)-linked chains from the bifurcose branch-point residue. Some of the (2----6)-linked units that slowly accumulate in oligomers may arise in vivo from selective hydrolysis, by fructan exohydrolases, of (2----1)-linked terminal units at branch point residues rather than by the action of (2----6)-specific synthases. Limited hydrolysis by specific exohydrolases in vitro coupled with separation of the oligomeric products constitutes an effective approach to the sequence analysis of complex oligosaccharides.

Rate of accumulation of fructan oligomers in wheat seedlings (Triticum aestivum L.) during the early stages of chilling treatment.

The accumulation of soluble carbohydrates was studied during chilling of 2-week-old wheat (Triticum aestivum L.) seedlings. The extracts were analysed using a C18 HPLC method separating hexoses, sucrose and a wide range of oligofructan. Isomers were recognized and their content determined. The amounts of hexoses, sucrose and the trisaccharides 1-kestose and 6-kestose increased at the beginning of the treatment. The accumulation rate of 1-kestose, the trisaccharide generally considered to be the precursor of fructan, increased ten fold with some delay. Heavier fructan accumulated even later. In leaf blades, 1-3 molecular species with a degree of polymerization from 4 to 8 were detected. Five of these accounted for 90% of this pool at day 5. In leaf bases the amount of carbohydrates was lower but more species of fructan were detected. The sequence of accumulation was the same as in the blades. These results are discussed in relation to current models ot fructan synthesis.