An Overview of the Physiology and Biochemistry of N Reserves Mobilization in Forage Species

Recent works focusing on the physiological and biochemical events associated to perennial forages sustainability (re-growth after defoliation, winter survival), provided evidence that vegetative storage proteins (VSPs, ca proteins stored in remaining organs and specifically mobilized) were key organic compounds for shoot regrowth. Attempts to elucidate environmental conditions involved in VSPs deposit ability of different species are under progress today. In contrast, mechanisms of spring- or cut-induced proteolysis are investigated at a lesser extend. This contribution is an opportunity to summarize our knowledge of N-reserves mobilization and to set the question of VSPs breakdown regulation

Effects of Photoperiod, Low Temperature and N Nutrition on VSP Accumulation in Taproot of Alfalfa

In Medicago sativa L., vegetative storage protein (VSP), specifically accumulated in taproot, are strongly involved in nitrogen storage. How the accumulation of such VSPs is regulated remains largely unknown. Experiments were designed with non-nodulated alfalfa to determine if length of the photoperiod, a decrease of temperature, or high availability of mineral nitrogen may induce the accumulation of VSPs. 15N labelling was used to quantify nitrogen uptake and its further relative translocation within the plant while VSPs accumulation was analysed by ELISA quantification. Results showed that environmental factors such as shortening daylength or low temperature changed biomass allocation within the plant by reducing shoot growth. As a consequence, short days promoted the relative N allocation to taproot whereas VSP accumulation showed a higher trend. On the other hand, low temperature, changes in N source or availability in the nutrient solution, may lead to a higher influx of nitrogen and a higher soluble protein relative concentration in taproot while VSP abundance remained low

Seasonal Fluctuations of Vegetative Storage Proteins and Starch Concentrations in Stolons of Trifolium Repens L.

The seasonal pattern of nitrogen, starch and vegetative storage protein (VSP) concentrations was studied in the stolons of two Trifolium repens L. genotypes (cv Aran and Rivendel). Maximum concentrations of starch were found in summer months; its hydrolysis occurred in winter, at the time, where VSP and nitrogen were accumulated. The decrease of nitrogen and VSP concentrations occurred during spring, and an inverse relationship was found between VSP concentrations in stolons and mean temperatures. The causal implications of starch and VSP availability on spring regrowth potential are discussed in relation with regulatory mechanism inducing VSP synthesis

Reserve mobilization during regrowth after cutting of forage species: quantification and physiological mechanisms in ryegrass and lucerne

Time course of remobilisation of reserves was studied during the regrowth of Loli11m perem1e and Medicago sativa by 15N labelling, carbohydrate analyses and by measuring time course of several enzymes activities. When photosynthesis decreased during early regrowth, and consequently reduced availability of carbon skeletons within the plant, both nitrogen (N) uptake (lolium perene) and N2 fixatlon (Medicago saliva) decreased. Nilrogenase, as well as nitrate reductase activities, decreased. Reduction of external N assimilation was compensated for by intensive mobilization of endogenous N in roots of lucerne and in stubble of ryegrass. In the same manner, some of the carbon (C) reserves of organs remaining after defoliation were translocated to regrowing shoals. Activities of hydrolytic enzymes such as fructan-exo-hydrolase or proteinase therefore increased after defoliation in all remaining organs which acted as source organs. The second period of regrowth corresponded with an increase in carbohydrate synthesis, and was associated with (i) reconstitution of N and C reserves concomitant with a decrease of hydrolytic activities, (ii) increases of mineral N uptake and N2 fixation associated with higher enzyme activities

Root Nitrogen Cycling and Alfalfa Stress Tolerance

Our hypothesis is that certain root N pools are utilized preferentially during the early shoot regrowth. Our objective was to determine the relative contribution of crown N, root N, and specific root N pools to shoot regrowth after defoliation. We used 15N to follow N into and out of crowns, roots, and specific root N pools, to regrowing shoots after defoliation. The low molecular weight N pool (amino acids, inorganic N,...) acquired 15N rapidly within 2 d of N application. Movement of 15N into the protein-N and insoluble-N pools was delayed initially, but continued until 8 d after N application. Defoliation 30 d after 15N application resulted in N transfer from roots and crowns to regrowing shoots. All root N pools lost 15N label initially after defoliation, with a more extensive decline occurring for the protein-N and low molecular weight-N pools

Modelling Nitrogen Uptake in Winter Oilseed Rape by Using Influx Kinetics of Nitrate Transport Systems

A mechanistic model was proposed in order to predict nitrogen uptake by a culture of oilseed rape (Brassica napus L.), using independently measured characteristics of plants growing in hydroponic or under field conditions. Uptake kinetics of the different components (Constitutive and Inducible) of the Low and High Affinity Transport Systems of nitrate (CLATS, ILATS, CHATS and IHATS, respectively) were determined by 15NO3- labelling in controlled conditions. The use of kinetic equations of transport systems and the experimental field data from the INRA-Châlons rape databank allowed to model NO3- uptake during the plant growth cycle. The study of different factors such as root temperature, day/night cycle and ontogenetic stages on NO3- uptake rate has been undertaken in order to improve the model prediction. Model outputs show that the high affinity transport system (HATS) accounted for about 90 % of total NO3- uptake (20 and 70 % for CHATS and IHATS without fertilization, respectively). The low affinity transport system (LATS) accounted for a minor proportion of total N uptake, and its activity was restricted to the early phase of the growth cycle. However, N autumnal fertilization increased the duration of its contribution (from 67 to 100 days) to total N uptake

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