There is an increasing need for diagnostic tools that can assess the crop nitrogen (N) nutrition status during the growth cycle. In addition to the leaf chlorophyll (Chl) content, we proposed here ...the use of the leaf content of polyphenolics (Phen) as a potential indicator of crop N status. Because of their absorption features in the visible and in the UV part of the spectrum, both Chl and Phen can be measured by rapid and non-destructive optical methods. Therefore, we used two leaf-clip devices, the Minolta SPAD-502 for Chl, and the Dualex for Phen. The latter is a prototype (patent pending) that measures the UV absorbance of the leaf epidermis, which is related to the leaf Phen content. Dynamics of Phen and Chl were measured on the last fully developed leaves of two winter wheat cultivars subjected to different levels of N availability, from tillering to flowering, in 2001, 2002 and 2003. Both Phen and Chl contents were found to increase along the leaf, starting from the ligula, regardless of the stage of development. Both variables were highly correlated with the N concentration of leaves. The average Chl content of the leaf increased, and the average Phen content decreased, with the increased application of N to the field, irrespective of the growth stage, the cultivar and the year of experiment. Therefore, both Phen and Chl can be considered as probes of the crop N nutrition status. Still, the relationship between Chl and the nitrogen nutrition index (NNI), used as a reference indicator of N deficiency, was influenced by the growth stage, whereas the year of experiment affected the relationship between Phen and the NNI. We also propose the use of the simple Chl/Phen ratio as an indicator of leaf N content at the canopy level, for future application in precision agriculture. This ratio would alleviate, at least partially, the problem of gradients along leaves, and would even accentuate the differences among levels of crop N deficiencies because of the Chl and Phen inverse dependence on the crop N nutrition status.
Between flowering and maturity, leaf senescence reduces green leaf area while grains are filled from photosynthesis, nitrogen (N) uptake by roots and remobilisation from shoots. The question arises ...of possible effects of leaf senescence on photosynthesis, N uptake and remobilisation and their modification through genetic variations in senescence. To address this question, we compared three cultivars showing different behaviours. Tarro and Nicco are two modern hybrids presenting the same important grain sink but showing different time course of senescence, the former being normally senescent the latter “stay-green”. Déa hybrid, very well known by many previous experiments was the reference hybrid.
The purpose was to monitor precisely the carbon (C) and N repartition in the main organs of the plant in connection with the progress of leaf senescence (experiment 1) and to measure the changes created by variations of N supply and soil-climate conditions (experiment 2).
In experiment 1, time course of leaf senescence of the different leaf stages and weight of leaves, shoots and grains and N concentration of shoots and grain were measured weekly. In experiment 2, the same three cultivars were grown at limiting and non-limiting N supply in four locations in France during 2 years. At flowering, silage and harvest stage, dry matter and N concentration were measured in the main parts of the plant, and leaf senescence was evaluated at silage stage.
On soil well supplied with N, Tarro had an identical rate of leaf senescence than Nicco for leaves below the ear, but higher for leaves above the ear. After flowering, Nicco accumulated more biomass than Tarro, but kept a larger part of this total biomass in the stem. In contrast, Tarro accumulated a larger part of total biomass in the grain.
After flowering, N uptake was larger in Nicco than Tarro. Shoot N concentration decreased earlier and more completely in Tarro than Nicco, indicating a larger remobilisation to the grain. Though Tarro began and finished grain filling simultaneously with Nicco, it reached 70% of final grain weight 25
°C
d sooner than Nicco.
The senescence due to N stress differed from the physiological senescence of a senescent hybrid. The time course of the reproductive phase and the N repartition in the plant suggest that a threshold in grain filling or in shoot nitrogen decrease could be the internal signal triggering senescence of uppermost leaves.
A two-dimensional reference map of maize endosperm proteins was established, with the identification of 496 spots sorted in 15 functional categories. The three most abundant categories were ...metabolism, protein destination and protein synthesis. This map is the first step for investigating maize endosperm development.
We have established a proteome reference map for maize (
Zea mays L.) endosperm by means of two-dimensional gel electrophoresis and protein identification with LC–MS/MS analysis. This investigation focussed on proteins in major spots in a 4–7 p
I range and 10–100 kDa
M
r range. Among the 632 protein spots processed, 496 were identified by matching against the NCBInr and ZMtuc-tus databases (using the SEQUEST software). Forty-two per cent of the proteins were identified against maize sequences, 23% against rice sequences and 21% against
Arabidopsis sequences. Identified proteins were not only cytoplasmic but also nuclear, mitochondrial or amyloplastic. Metabolic processes, protein destination, protein synthesis, cell rescue, defense, cell death and ageing are the most abundant functional categories, comprising almost half of the 632 proteins analyzed in our study. This proteome map constitutes a powerful tool for physiological studies and is the first step for investigating the maize endosperm development.
One hundred to 120 maize recombinant inbred lines at the mature fourth leaf stage derived from F-2 and Io parental lines were grown in a glasshouse and were deprived of water for 9 days in order to ...detect pertinent markers of the physiological response to water stress which may be used for breeding. Carbohydrate metabolism QTLs were compared to photosynthesis gas exchange QTLs. The locations of these QTLs were further compared with those of morphological trait QTLs when water availability varied. The traits ranged from three enzyme activities (invertase, sucrose-P synthase, ADP glucose pyrophosphorylase) and hexose, sucrose, starch content to CO2 uptake and stomatal conductance, water status, leaf size, root/shoot ratio, and ABA (leaf, root and xylem sap). Four main results were obtained (1) only 14 % of QTLs were common to both drought and watered treatments, confirming the existence of stress specific chromosome regions, (2) the QTLs tended to form clusters, frequently consisting of QTLs from different classes (growth, photosynthesis, water status, carbohydrate metabolism and ABA), (3) carbohydrate metabolism trait QTLs were more frequently co-located with growth trait QTLs than photosynthesis related ones, especially in control conditions, (4) one co-location was observed between the three enzyme activities implied in sucrose and starch metabolism and a corresponding structural gene, which can be considered as a candidate gene for explaining part of the variability of each enzymatic trait (invertase, sucrose-P synthase, ADPglucose pyrophosphorylase). It is concluded that, carbohydrate metabolism provides valuable traits for understanding and improving maize responses to water stress.
The expression of invertases was analyzed in vegetative organs of well-watered and water-stressed maize (Zea mays) plants. Early changes in sucrose metabolism and in acid soluble invertase expression ...were observed in vegetative sink and source organs under mild water stress. The organ-specific induction of acid invertase activity was correlated with an increase in the Ivr2 gene transcripts and in the vacuolar invertase proteins. In addition diurnal changes in activity and Ivr2 transcripts for vacuolar invertase were noted in shoots. Hexoses (glucose and fructose) accumulated in all organs examined from water-stressed plants. In situ localization studies showed that glucose accumulation, vacuolar invertase activity, invertase protein, and the Ivr2 transcripts colocalized specifically in bundle sheath and vascular tissue cells of mature stressed leaf; in primary roots the stress-induced increase of Ivr2 transcripts was detected only in root tips. Based on these results different regulatory roles are proposed in sink and source organs for the stress induced Ivr2 vacuolar invertase.
ABSTRACT
The time course of the modifications induced by a mild water stress has been examined for photosynthesis and several traits of carbohydrate metabolism in adult leaves of two inbred maize ...lines of North American and European origins, respectively. An early response was a sharp increase of the acid soluble invertase activity in adult leaves, 3–4 d after initiation of water shortage. Accordingly, correlated accumulations of fructose, glucose and to a lesser extent sucrose were observed. In the most responsive genotype, invertase activity finally reached a value > 3 times larger than the control value. By contrast, sucrose phosphate synthase activity, measured either under saturating or limiting substrate conditions, was progressively reduced by 20–40% on the 5th day and by 50–80% on the 7th day, depending on the genotype. Leaf photosynthetic rate was affected at approximately the same time as carbohydrate metabolism and stomatal conductance. Leaf water status, as measured by relative water content, declined afterwards. For all the observed responses, the two genotypes behaved very differently.
Estimating the proportion of N remobilization and postsilking N uptake allocated to kernels may help to improve N-use efficiency in maize (Zea mays L.). In this study, we show theoretically and ...experimentally, that 15N labeling at the beginning of stem elongation can be used in the field to estimate the proportion of N remobilized from the vegetative parts to the kernels of maize by measuring 15N distribution only at maturity. In the same way, 15N labeling at silking allows a determination of the proportion of postsilking N uptake allocated to the kernels by measuring 15N distribution only at maturity. Two 1-yr experiments with three and four genotypes and one 2-yr experiment with testcross progenies from 66 recombinant inbred lines were developed. Nitrogen-15 labeling during vegetative growth provided an estimate of the proportion of N remobilized with a greater accuracy compared with the "balance" method, which computes the difference between the amount of N present in the stover at silking and the amount of N present in the stover at maturity. The validity of our 15N method is mainly based on the assumption that less than 15 to 20% of 15N is taken up after silking. The determination of the proportion of postsilking N uptake allocated to kernels requires assumptions that are more difficult to fulfill. Nitrogen-15 labeling in the field at the beginning of rapid stem growth appears to be a useful tool for studying the genetic variability of N remobilization using a large number of genotypes.
Plastic deformation in irradiated zirconium alloys occurs in a very heterogeneous manner at the grain scale by the clearing up of radiation-induced prismatic loops by gliding dislocations and the ...channeling of these dislocations inside narrow bands of the grain. A statistical TEM investigation of this mechanism has been performed on neutron irradiated recrystallized Zr alloys tested at 350 °C. Due to the strong anisotropy of plastic deformation, different loading conditions have been investigated. It is shown that for transverse tensile tests and closed end burst tests, only basal channels are observed, whereas for axial tensile tests, prismatic and pyramidal channels are observed. This phenomenon can be understood in terms of texture and interactions between dislocations and irradiation induced loops. From Schmid factor calculations, we have also been able to prove that irradiation leads to a higher increase of critical resolved shear stresses for prismatic and pyramidal slip systems than for basal slip system.
The Nicotiana sylvestris mutant, CMS, lacks the mitochondrial gene nad7 and functional complex I, and respires using low-affinity NADH (alternative) mitochondrial dehydrogenases. Here, we show that ...this adjustment of respiratory pathways is associated with a profound modification of foliar carbon-nitrogen balance. CMS leaves are characterized by abundant amino acids compared to either wild-type plants or CMS in which complex I function has been restored by nuclear transformation with the nad7 cDNA. The metabolite profile of CMS leaves is enriched in amino acids with low carbon/nitrogen and depleted in starch and 2-oxoglutarate. Deficiency in 2-oxoglutarate occurred despite increased citrate and malate and higher capacity of key anaplerotic enzymes, notably the mitochondrial NAD-dependent isocitrate dehydrogenase. The accumulation of nitrogen-rich amino acids was not accompanied by increased expression of enzymes involved in nitrogen assimilation. Partitioning of ¹⁵N-nitrate into soluble amines was enhanced in CMS leaf discs compared to wild-type discs, especially in the dark. Analysis of pyridine nucleotides showed that both NAD and NADH were increased by 2-fold in CMS leaves. The growth retardation of CMS relative to the wild type was highly dependent on photoperiod, but at all photoperiod regimes the link between high contents of amino acids and NADH was observed. Together, the data provide strong evidence that (1) NADH availability is a critical factor in influencing the rate of nitrate assimilation and that (2) NAD status plays a crucial role in coordinating ammonia assimilation with the anaplerotic production of carbon skeletons.
Plant ADP-glucose pyrophosphorylase (AGP) is a heterotetrameric enzyme composed of two large and two small subunits. Here, we report the structures of the maize (Zea mays) genes encoding AGP small ...subunits of leaf and endosperm. Excluding exon 1, protein-encoding sequences of the two genes are nearly identical. Exon 1 coding sequences, however, possess no similarity. Introns are placed in identical positions and exhibit obvious sequence similarity. Size differences are primarily due to insertions and duplications, hallmarks of transposable element visitation. Comparison of the maize genes with other plant AGP small subunit genes leads to a number of noteworthy inferences concerning the evolution of these genes. The small subunit gene can be divided into two modules. One module, encompassing all coding information except that derived from exon 1, displays striking similarity among all genes. It is surprising that members from eudicots form one group, whereas those from cereals form a second group. This implies that the duplications giving rise to family members occurred at least twice and after the separation of eudicots and monocot cereals. One intron within this module may have had a transposon origin. A different evolutionary history is suggested for exon 1. These sequences define three distinct groups, two of which come from cereal seeds. This distinction likely has functional significance because cereal endosperm AGPs are cytosolic, whereas all other forms appear to be plastid localized. Finally, whereas barley (Hordeum vulgare) reportedly employs only one gene to encode the small subunit of the seed and leaf, maize utilizes the two genes described here.
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