Photosynthesis, N(2) fixation and taproot reserves during the cutting regrowth cycle of alfalfa under elevated CO(2) and temperature.

Future climatic conditions, including rising atmospheric CO(2) and temperature may increase photosynthesis and, consequently, plant production. A larger knowledge of legume performance under the predicted growth conditions will be crucial for safeguarding crop management and extending the area under cultivation with these plants in the near future. N(2) fixation is a key process conditioning plant responsiveness to varying growth conditions. Moreover, it is likely to increase under future environments, due to the higher photosynthate availability, as a consequence of the higher growth rate under elevated CO(2). However, as described in the literature, photosynthesis performance is frequently down-regulated (acclimated) under long-term exposure to CO(2), especially when affected by stressful temperature and water availability conditions. As growth responses to elevated CO(2) are dependent on sink-source status, it is generally accepted that down-regulation occurs in situations with insufficient plant C sink capacity. Alfalfa management involves the cutting of shoots, which alters the source-sink relationship and thus the photosynthetic behaviour. As the growth rate decreases at the end of the pre-cut vegetative growth period, nodulated alfalfa plants show photosynthetic down-regulation, but during regrowth following defoliation, acclimation to elevated CO(2) disappears. The shoot harvest also leads to a drop in mineral N uptake and C translocation to the roots, resulting in a reduction in N(2) fixation due to the dependence on photosynthate supply to support nodule function. Therefore, the production of new shoots during the first days following cutting requires the utilization of reduced C and N compounds that have been stored previously in reserve organs. The stored reserves are mediated by phytohormones such as methyl jasmonate and abscisic acid and in situations where water stress reduces shoot production this potentially enables the enhancement of taproot protein levels in nodulated alfalfa, which may lead to these plants being in better condition in the following cut/regrowth cycle. Furthering our knowledge of legume performance under predicted climate change conditions will be crucial for the development of varieties with better adaptation that will achieve greater and more efficient production values. Furthermore, for this purpose it will be necessary to improve existing methodologies and create new ones for phenotype characterization. Such knowledge will provide key information for future plant breeding programs.

Is the remobilization of S and N reserves for seed filling of winter oilseed rape modulated by sulphate restrictions occurring at different growth stages?

How the remobilization of S and N reserves can meet the needs of seeds of oilseed rape subject to limitation of S fertilization remains largely unclear. Thus, this survey aims to determine the incidence of sulphate restriction [low S (LS)] applied at bolting [growth stage (GS) 32], visible bud (GS 53), and start of pod filling (GS 70) on source-sink relationships for S and N, and on the dynamics of endogenous/exogenous S and N contributing to seed yield and quality. Sulphate restrictions applied at GS 32, GS 53, and GS 70 were annotated LS(32), LS(53), and LS(70). Long-term (34)SO(4)(2-) and (15)NO(3)(-) labelling was used to explore S and N partitioning at the whole-plant level. In LS(53), the sulphur remobilization efficiency (SRE) to seeds increased, but not enough to maintain seed quality. In LS(32), an early S remobilization from leaves provided S for root, stem, and pod growth, but the subsequent demand for seed development was not met adequately and the N utilization efficiency (NUtE) was reduced when compared with high S (HS). The highest SRE (65 ± 1.2% of the remobilized S) associated with an efficient foliar S mobilization (with minimal residual S concentrations of 0.1-0.2% dry matter) was observed under LS(70) treatment, which did not affect yield components.

Effect of mineral sulphur availability on nitrogen and sulphur uptake and remobilization during the vegetative growth of Brassica napus L.

Because it has a high demand for sulphur (S), oilseed rape is particularly sensitive to S limitation. However, the physiological effects of S limitation remain unclear, especially during the rosette stage. For this reason a study was conducted to determine the effects of mineral S limitation on nitrogen (N) and S uptake and remobilization during vegetative growth of oilseed rape at both the whole-plant and leaf rank level for plants grown during 35 d with 300 microM (34)SO(4)(2-) (control plants; +S) or with 15 microM (34)SO(4)(2-) (S-limited plants; -S). The results highlight that S-limited plants showed no significant differences either in whole-plant and leaf biomass or in N uptake, when compared with control plants. However, total S and (34)S (i.e. deriving from S uptake) contents were greatly reduced for the whole plant and leaf after 35 d, and a greater redistribution of endogenous S from leaves to the benefit of roots was observed. The relative expression of tonoplast and plasmalemma sulphate transporters was also strongly induced in the roots. In conclusion, although S-limited plants had 20 times less mineral S than control plants, their development remained surprisingly unchanged. During S limitation, oilseed rape is able to recycle endogenous S compounds (mostly sulphate) from leaves to roots. However, this physiological adaptation may be effective only over a short time scale (i.e. vegetative growth).

Remobilization of leaf S compounds and senescence in response to restricted sulphate supply during the vegetative stage of oilseed rape are affected by mineral N availability.

The impact of sulphur limitation on the remobilization of endogenous S compounds during the rosette stage of oilseed rape, and the interactions with N availability on these processes, were examined using a long-term (34)SO(4)(2-) labelling method combined with a study of leaf senescence progression (using SAG12/Cab as a molecular indicator) and gene expression of the transporters, BnSultr4;1 and BnSultr4;2, involved in vacuolar sulphate efflux. After 51 d on hydroponic culture at 0.3 mM (34)SO(4)(2-) (1 atom% excess), the labelling was stopped and plants were subject for 28 d to High S-High N (HS-HN, control), Low S-High N (LS-HN) or Low S-Low N (LS-LN) conditions. Compared with the control, LS-HN plants showed delayed leaf senescence and, whilst the shoot growth and the foliar soluble protein amounts were not affected, S, (34)S, and SO(4)(2-) amounts in the old leaves declined rapidly and were associated with the up-regulation of BnSultr4;1. In LS-LN plants, shoot growth was reduced, leaf senescence was accelerated, and the rapid S mobilization in old leaves was accompanied by decreased (34)S and SO(4)(2-), higher protein mobilization, and up-regulation of BnSultr4;2, but without any change of expression of BnSultr4;1. The data suggest that to sustain the S demand for growth under S restriction (i) vacuolar SO(4)(2-) is specifically remobilized in LS-HN conditions without any acceleration of leaf senescence, (ii) SO(4)(2-) mobilization is related to an up-regulation of BnSultr4;1 and/or BnSultr4;2 expression, and (iii) the relationship between sulphate mobilization and up-regulation of expression of BnSultr4 genes is specifically dependent on the N availability.

Developmental changes in shoot N dynamics of lucerne (Medicago sativa L.) in relation to leaf growth dynamics as a function of plant density and hierarchical position within the canopy.

Shoot N concentration in plants decreases as they get bigger, due to the fact that N accumulates less rapidly than dry matter in plants during the plant growth process, leading to an allometric relationship between shoot N content (N(sh)) and shoot mass (W(sh)): N(sh)=a(W(sh))b. The results obtained on lucerne plants growing either under controlled low density conditions or in dense stands under field conditions show that the value of the allometric coefficient b that represents the ratio between the relative N accumulation rate in shoots [dN(sh)/(N(sh)dt)] and the relative growth rate [dW(sh)/(W(sh)dt)], decreases from 0.88 for a low plant density to 0.72 for a dense stand. Therefore, the fractional increase of shoot N per unit of shoot dry matter is lower when plants are in competition for light in dense canopies. This decrease can be entirely explained by the parallel decline in the leaf area per unit of shoot mass. Thus, a remarkably constant linear relationship can be established between N(sh) and leaf area (LA): N(sh)=1.7 g m(-2) LA, regardless of the conditions (low versus high density, controlled versus field conditions). Moreover, in a field dense stand, the comparison of plants with contrasting positions between the top and the bottom of the canopy (dominant, intermediate or suppressed plants), also shows that the difference in N(sh) at similar shoot mass is explained by the proportion of leaf mass to shoot mass. These data support the idea that leaf growth drives the dynamics of shoot N accumulation. These results also indicate that competition for light among individual plants within a dense canopy induces developmental changes in plant morphology (leaf:stem ratio) that explain the differences observed in shoot N concentration. This last observation could be extrapolated to multi-specific plant stands. Therefore, the sharing of N resources among plant species could partially be the result of the sharing of light within the canopy.

Influence of initial organic N reserves and residual leaf area on growth, N uptake, N partitioning and N storage in alfalfa (Medicago sativa) during post-cutting regrowth.

BACKGROUND AND AIMS: The influence of initial residual leaf area and initial N reserves on N uptake, final N distribution, and yield in alfalfa regrowing after cutting, were studied. METHODS: The effects of two levels of initial residual leaf area (plants cut to 15 cm, with (L+) or without (L-) their leaves) and two initial levels of N status [high N (HN) or low N (LN)] on growth, N uptake and N partitioning, allocation and storage after 29 d of post-cutting regrowth were analysed. KEY RESULTS: During most of the regrowth period (8-29 d after the initial harvest), HN and L+ plants had higher net N uptake rates than LN and L- plants, respectively, resulting in a greater final mineral N uptake for these treatments. However, the final partitioning of exogenous N to the regrowing shoots was the same for all treatments (67 % of total exogenous N on average). Final shoot growth, total plant N content, and N allocation to the different taproot N pools were significantly lower in plants with reduced initial leaf area and initial N reserve status. CONCLUSIONS: Although both initial residual leaf area and initial N reserves influenced alfalfa regrowth, the residual leaf area had a greater effect on final forage production and N composition in the taproot, whereas the N uptake rate and final total N content in plant were more affected by the initial N reserve status than by the residual leaf area. Moreover, N storage as proteins (especially as vegetative storage proteins, rather than nitrate or amino acids) in the taproot allowed nitrate uptake to occur at significant rates. This suggests that protein storage is not only a means of sequestering N in a tissue for further mobilization, utilization for growth or tissue maintenance, but may also indirectly influence both N acquisition and reduction capacities.

A rapid and reliable method for NO quantification and 15NO/14NO determination using isotope ratio mass spectrometry: an application for the detection of NO synthesis in propionibacteria.

For the last decade, numerous studies have focused on the positive or toxic effects of nitric oxide (NO) in procaryotic and eucaryotic cells. This gas has fundamental roles in neurotransmission, vasodilatation, cytotoxicity, and intestinal motility. The ability to produce NO by intestinal microflora or probiotic bacteria is unknown. In this preliminary study, we present a rapid and reproducible procedure for NO quantification and 15NO/14NO determination (based on the reaction between nitrite and acidic potassium iodide) by isotope ratio mass spectrometry. Using this method, we have demonstrated for the first time in vitro production of NO by a dietary bacterium (Propionibacterium acidipropionici, Pa 1) under anaerobic culture conditions. Using different sources of nitrogen, we have clearly shown that propionibacteria can synthesize NO from reduction of nitrate or nitrite. In our experimental conditions, NO synthase was not involved in NO production by propionibacteria.

Nitrogen and Carbon Flows Estimated by 15N and 13C Pulse-Chase Labeling during Regrowth of Alfalfa.

The flow of 15N and 13C from storage compounds in organs remaining after defoliation (sources) to regrowing tissue (sinks), and 13C losses through root or shoot respiration were assessed by pulse-chase labeling during regrowth of alfalfa (Medicago sativa L.) following shoot removal. A total of 73% of labeled C and 34% of labeled N were mobilized in source organs within 30 d. Although all of the 15N from source organs was recovered in the regrowing tissue, much of the 13C was lost, mainly as CO2 respired from the root (61%) or shoot (8%), and was found to a lesser extent in sink tissue (5%). After 3, 10, or 30 d of regrowth, 87, 66, and 52% of shoot N, respectively, was derived from source tissue storage compounds; the rest resulted from translocation of fixed N2. Overall results suggest that most shoot C was linked to photosynthetic activity rather than being derived from mobilization of stored C in source organs. Furthermore, isotopic analysis of different chemical fractions of plant tissue suggests that between 14 and 58% of the shoot C derived from source tissues was linked to the mobilization of N compounds, not carbohydrates.