Investigating tree and fruit growth through functional-structural modelling: implications of carbon autonomy at different scales.

BACKGROUND AND AIMS: Many experimental studies assume that some topological units are autonomous with regard to carbon because it is convenient. Some plant models simulate carbon allocation, employing complex approaches that require calibration and fitted parameters. For whole-tree canopy simulations, simpler carbon allocation models can provide useful insights. METHODS: We propose a new method for simulating carbon allocation in the whole tree canopy considering various scales of carbon autonomy, i.e. branchlets, branches, limbs, and no autonomy. This method was implemented in a functional-structural plant model of growth of individual organs for studying macadamia tree growth during one growing season. KEY RESULTS: This model allows the simulation of various scales of carbon autonomy in a simple tree canopy, showing organ within-tree variability according to the scale of autonomy. Using a real tree canopy, we observed differences in growth variability within the tree and in tree growth, with several scales of carbon autonomy. The simulations that assumed autonomy at branch scale, i.e. 2-year-old wood, showed the most realistic results. CONCLUSIONS: Simulations using this model were employed to investigate and explain aspects of differences in carbon autonomy between trees, organ growth variability, competition between shoot and fruit growth, and time of autonomy.

Linking water stress effects on carbon partitioning by introducing a xylem circuit into L-PEACH.

BACKGROUND AND AIMS: Many physiological processes such as photosynthesis, respiration and transpiration can be strongly influenced by the diurnal patterns of within-tree water potential. Despite numerous experiments showing the effect of water potential on fruit-tree development and growth, there are very few models combining carbohydrate allocation with water transport. The aim of this work was to include a xylem circuit into the functional-structural L-PEACH model. METHODS: The xylem modelling was based on an electrical circuit analogy and the Hagen-Poisseuille law for hydraulic conductance. Sub-models for leaf transpiration, soil water potential and the soil-plant interface were also incorporated to provide the driving force and pathway for water flow. The model was assessed by comparing model outputs to field measurements and published knowledge. KEY RESULTS: The model was able to simulate both the water uptake over a season and the effect of different irrigation treatments on tree development, growth and fruit yield. CONCLUSIONS: This work opens the way to a new field of modelling where complex interactions between water transport, carbohydrate allocation and physiological functions can be simulated at the organ level and describe functioning and behaviour at the tree scale.

A self-induction method to produce high quantities of recombinant functional flavo-leghemoglobin reductase.

Ferric leghemoglobin reductase (FLbR) is able to reduce ferric leghemoglobin (Lb3+) to ferrous (Lb2+) form. This reaction makes Lb functional in performing its role since only reduced hemoglobins bind O2. FLbR contains FAD as prosthetic group to perform its activity. FLbR-1 and FLbR-2 were isolated from soybean root nodules and it has been postulated that they reduce Lb3+. The existence of Lb2+ is essential for the nitrogen fixation process that occurs in legume nodules; thus, the isolation of FLbR for the study of this enzyme in the nodule physiology is of interest. However, previous methods for the production of recombinant FLbR are inefficient as yields are too low. We describe the production of a recombinant FLbR-2 from Escherichia coli BL21(DE3) by using an overexpression method based on the self-induction of the recombinant E. coli. This expression system is four times more efficient than the previous overexpression method. The quality of recombinant FLbR-2 (based on spectroscopy, SDS-PAGE, IEF, and native PAGE) is comparable to that of the previous expression system. Also, FLbR-2 is purified near to homogeneity in only few steps (in a time scale, the full process takes 3 days). The purification method involves affinity chromatography using a Ni-nitrilotriacetic acid column. Resulting rFLbR-2 showed an intense yellow color, and spectral characterization of rFLbR-2 indicated that rFLbR-2 contains flavin. Pure rFLbR-2 was incubated with soybean Lba and NADH, and time drive rates showed that rFLbR-2 efficiently reduces Lb3+.

Use of recombinant iron-superoxide dismutase as a marker of nitrative stress.

Superoxide dismutases (SODs; EC 1.15.1.1) are a group of metalloenzymes which are essential to protect cells under aerobic conditions. In biological systems, it has been reported that SODs and other proteins are susceptible to be attacked by peroxynitrite (ONOO(-)) which can be originated from the reaction of nitric oxide with superoxide radical. ONOO(-) is a strong oxidant molecule capable of nitrating peptides and proteins at the phenyl side chain of the tyrosine residues. In the present work, bovine serum albumin (BSA) and recombinant iron-superoxide dismutase from the plant cowpea (Vu_FeSOD) are used as target molecules to estimate ONOO(-) production. The method employs the compound SIN-1, which simultaneously generates *NO and O(2)(-) in aerobic aqueous solutions. First, assay conditions were optimized incubating BSA with different concentrations of SIN-1, and at a later stage, the effect on the tyrosine nitration and catalytic activity of Vu_FeSOD was examined by in-gel activity and spectrophotometric assays. Both BSA and Vu_FeSOD are nitrated in a dose-dependent manner, and, at least in BSA nitration, the reaction seems to be metal catalyzed.