Estimation of Carbon and Nitrogen Allocation during Stalk Elongation by C and N Tracing in Zea mays L

Zea mays L. (cv Dea) plants grown to the stage of stalk elongation, were allowed to assimilate ¹³CO₂ and ¹⁵N-nitrates from 45 to 53 days after sowing. Isotopic abundances in labeled nutrients were slightly enriched compared to natural abundances. The new C in plant was acropetally distributed and the new N was preferentially accumulated in the sheath and stalk in the medium region. C input was 25-fold higher than N input. The new C in total plant C was 20%, whereas it was 10% for N. The stalk acted as a major sink because it accumulated, respectively, 27.5 and 47.5% of the C and N inputs. The new C in soluble carbohydrates was 76% in growing organs (upper stalk) and only 39% in source leaves, whereas it was 43% and 13% in starch, respectively. New N in nitrates+amino-acids spanned in the range from 20% (leaf) to 50% (stalk). New C and N in soluble proteins were, respectively, 13.4 and 3.8% in leaves, 8.8 and 9.6% in stalk, and 8.7 and 14.3% in roots. In the middle stalk and leaves, the proteins and carbohydrates represent an equivalent C and N source for remobilization.     

Nitrogen metabolism in the stalk tissue of maize

During ear development in maize (Zea mays L.), nitrogenous compounds are translocated from vegetative organs to the kernels. At anthesis, the stalk contains approximately 40% of the total plant N, and contributes 45% of the N remobilized to the ear. Therefore, the stalk has an important function as a temporary reservoir for N. Little is known of the metabolism of maize stalks, and this paper describes initial studies of enzymes of N metabolism. High in vitro activity of glutamine synthetase (GS) in maize stalk samples throughout ear development contrasted with a peak in activity of glutamate synthase soon after anthesis and negligible nitrate reductase. With fresh sections of stalk tissue collected at anthesis, ¹⁵N-feeding experiments confirmed high GS and low nitrate reductase activities. Two isoforms of GS were separated from extracts from stalk tissue: GS1, the cytoplasmic form, increased to maximum levels at 2 weeks postanthesis and remained fairly high thereafter; whereas the plastidic form, GS2, declined progressively during kernel development. Western blot analysis confirmed the presence of constantly high levels of GS protein after anthesis. The levels of GS proteins decreased after transfer of N-starved, hydroponically grown plants to N-rich conditions in order to restrict remobilization of N. In contrast, transfer of plants grown under abundant N conditions to N-free medium, which encourages N remobilization, resulted in a relative increase in GS protein. Because glutamine is the major form of N transported in maize, the results indicate that GS, specifically the GS1 isoform, has a central role in the remobilization on nitrogenous compounds from the stalk to the ear.     

A proposed role of zein and glutelin as N sinks in maize

Zea mays grown with high levels of N fertilizer transports more sucrose into kernels than with low N. Sucrose translocation was greatest in genotypes with the highest capacity to deposit nitrogenous compounds as zein and glutelin in the kernel. These two proteins combined contain about 80% of the total N in the kernel and about 60% of the total N in the plant at maturity. They appear to serve as a functional N sink for the deposition of nitrogenous compounds. As the N sink capacity increases with additional available N fertilizer, more sucrose is transported into the kernel, resulting in increased kernel weight and grain yield. Zein functions as a more dynamic N sink than glutelin because the synthesis of zein is readily manipulated by N fertilization and genetic means. Increases in N deposition in the normal endosperm induced by N fertilizer are confined primarily to zein. Early termination of zein accumulation in the opaque-2 mutant results in a reduction of sucrose movement into kernels. By using plants heterozygous for normal and opaque-2 in these studies, interplant variability was eliminated and the hypothesis relating the kernel N sink capacity to productivity was strengthened.     

Utilization of Previously Accumulated and Concurrently Absorbed Nitrogen during Reproductive Growth in Maize : Influence of Prolificacy and Nitrogen Source

A prolific maize (Zea mays L.) genotype was grown to physiological maturity under greenhouse conditions to examine the effects of reproductive sink demand on (a) the remobilization of N accumulated during vegetative growth, and (b) the partitioning of N accumulated concurrent with ear development. One- and two-eared plants were treated with either a NO₃⁻ or NH₄⁺ source of ¹⁵N-labeled N during reproductive growth. Plants with two ears enhanced grain production, N remobilization from the stalk and roots, and N translocation to the grain from concurrently assimilated N. But, remobilization of leaf-N was unaffected by ear number. In addition, N uptake and total dry matter accumulation during the reproductive period were also unaffected, although P uptake was greater in the two-eared plants. Less than 15% of the total K⁺ uptake was accumulated after silking while during this time more than 40% of the total N and more than 50% of the total P were absorbed. The data also indicate that with NO₃⁻ nutrition, internal recirculation of K⁺ between shoots and roots may play a prominent role in the transport of nitrogenous solutes during grain development. N source had no effect on dry matter production and N uptake of both one- and two-eared plants. However, slightly greater partitioning of labeled-N from the NH₄⁺ source to the grain was observed in the two-eared plants.     

Remobilization patterns of C and N in soybeans with different sink-source ratios induced by various night temperatures

The effects of increased sink-source ratios, induced by elevating night temperatures, on remobilization of ¹⁴C-assimilates and N within field-grown soybeans (Glycine max [L.] Merr.) was investigated from preflowering to maturity. Raising the mean minimum night temperature for the entire growing season from 10 (check, uncontrolled) to 16°C increased seed growth without appreciable effect on final leaf area. Increasing this temperature to 24°C increased seed growth and reduced final leaf area. Leaves, stems, petioles, and pods acted as intermediate storage sites for ¹⁴C assimilates. Only plants with higher night temperatures remobilized some of the stored assimilates during the period of rapid seed growth. Even the seeds in the 24°C plants with the largest sink-source ratios did not utilize all the C-assimilates potentially available for remobilization. Nitrogen was readily remobilized from petioles, stems, and pods of all treatments as early as the beginning of seed development, but from the leaves only during late seed-filling. However, only plants with elevated night temperatures tended to remobilize all of the available N from vegetative tissues and pods. We concluded that a larger portion of stored assimilates may be remobilized to the seed if a strong seed sink can be sustained. It also appeared that with increasing sink-source ratios, N shortage might limit seed yield before a lack of C-assimilates would. A proposed model for soybean assimilate demand, distribution, partitioning, and remobilization is presented.     

Interaction of carbon and nitrogen metabolism in the productivity of maize

Five maize (Zea mays L.) hybrids, FS854, B73 × Mo17, B84 × Mo17, B73 × B77, and P3382, grown under field conditions, were sampled at intervals during the grain-filling period. Plants were subdivided into stalks (including sheaths), leaves, and kernels. These parts were assayed for dry weight, reduced nitrogen, and extractable nonstructural carbohydrates. The duration and rates of net nitrate reduction and photosynthesis were approximated by the changes over time in the accumulation of reduced nitrogen and dry weight by the plant (total, above ground), respectively.

Data on the accumulation of reduced nitrogen and dry weight by the plant show that decreases in nitrate reduction preceded (in time and extent for four of the hybrids and in extent for FS854) decreases or cessation of photosynthesis. FS854 continued to accumulate reduced nitrogen and dry matter throughout the grain-filling period.

The patterns of change in stalk carbohydrate and reduced nitrogen during the early stages of ear development show the stalk serves as a storage reservoir and that these reserves were remobilized during the final stages of grain development. The marked increase and maintenance of dry weight and carbohydrate content of stalks until 34 days after anthesis, shows the capacity of the leaves to produce photosynthate through the first half of the grain-filling period exceeds the needs of the ear and/or the transport system. In contrast, stalk nitrogen content shows a slight increase up to 12 days after anthesis and decreases continually thereafter. Leaf nitrogen was lost continuously throughout grain development. The potential capacity of the plant to supply newly reduced nitrogen was inadequate to support initiation and early development of the kernels without remobilization of vegetative nitrogen. Of the two hybrids having delayed leaf senescence, FS854 with its initially higher concentration and content of reduced nitrogen in the stalk, initiated and developed a bigger ear than P3382, which had lower levels of stalk nitrogen.

Three of the five hybrids had `near linear' rates of accumulation of kernel dry weight, whereas none of the hybrids had linear rates of gain in kernel nitrogen. All hybrids had maximum or near maximum rates of gain of kernel nitrogen between 26 and 34 days after anthesis and a marked reduction (41-52%) of rates in the following sampling interval. These decreases are concurrent with decreases in rates of nitrate reduction (nitrogen accumulation) by the whole plant for four of the hybrids and with decreases in remobilization of nitrogen from the vegetation of FS854. Data for the ratio of rates of accumulation of dry weight/reduced nitrogen by the kernels versus time after anthesis, show that the accumulation of dry weight and reduced nitrogen are independent of each other. The variations in the ratio values appear best related to variations in the availability of nitrogen from the vegetation.

     

C and N allocation in soil under ryegrass and alfalfa estimated by 13C and 15N labelling

Background and Aims

Below-ground translocated carbon (C) released as rhizodeposits is an important driver for microbial mobilization of nitrogen (N) for plants. We investigated how a limited substrate supply due to reduced photoassimilation alters the allocation of recently assimilated C in plant and soil pools under legume and non-legume species.

Methods

A non-legume (Lolium perenne) and a legume (Medicago sativa) were labelled with ¹⁵N before the plants were clipped or shaded, and labelled twice with ¹³CO₂ thereafter. Ten days after clipping and shading, the ¹⁵N and ¹³C in shoots, roots, soil, dissolved organic nitrogen (DON) and carbon (DOC) and in microbial biomass, as well as the ¹³C in soil CO₂ were analyzed.

Results

After clipping, about 50 % more ¹³C was allocated to regrowing shoots, resulting in a lower translocation to roots compared to the unclipped control. Clipping also reduced the total soil CO₂ efflux under both species and the ¹³C recovery of soil CO₂ under L. perenne. The ¹⁵N recovery increased in the shoots of M. sativa after clipping, because storage compounds were remobilized from the roots and/or the N uptake from the soil increased. After shading, the assimilated ¹³C was preferentially retained in the shoots of both species. This caused a decreased ¹³C recovery in the roots of M. sativa. Similarly, the total soil CO₂ efflux under M. sativa decreased more than 50 % after shading. The ¹⁵N recovery in plant and soil pools showed that shading has no effect on the N uptake and N remobilization for L. perenne, but, the ¹⁵N recovery increased in the shoot of M. sativa.

Conclusions

The experiment showed that the dominating effect on C and N allocation after clipping is the need of C and N for shoot regrowth, whereas the dominating effect after shading is the reduced substrate supply for growth and respiration. Only slight differences could be observed between L. perenne and M. sativa in the C and N distribution after clipping or shading.     

Effects of N-deficiency and timing of N supply on the recovery and distribution of labeled 15N in contrasting maize hybrids

Little information exists on the pattern of nitrogen (N) uptake, remobilization and N use efficiency (NUE) in Leafy and stay-green (SG) maize (Zea mays L.) genotypes. A pot experiment was conducted under controlled nutrition and growing conditions to determine the response of Leafy and SG maize genotypes to different levels of N-deficiency and timing of N supply. Three contrasting maize hybrids, Pioneer 3905 (a conventional hybrid with moderate SG characteristics), Pioneer 39F06 Bt (with a high score of SG trait) and Maizex LF850-RR (with a Leafy trait) were grown in 6 L plastic pots. Five different N treatments [no supply of N until V8 (N₁), no supply of N after V8 (N₂), no supply of N after silking (N₃), no supply of N beyond 3 weeks after silking (N₄), and continuous N supply from emergence to physiological maturity (N₅; standard check)] were imposed through modified Hoagland solution applied manually. Labeled ¹⁵N of 5% ¹⁵N₂–NH₄NO₃fertilizer was applied at 3 g per pot at the start of each schedule N treatment. Total amounts of N applied in different treatments were 3.13, 1.32, 1.90, 2.63 and 3.40 g, respectively in N₁, N₂, N₃, N₄ and N₅. Dry matter, N concentration, ¹⁵N (atom% enrichment) and NUE were determined in roots, stalk, leaves and grains at crop maturity. The three contrasting hybrids did not differ in grain yield, total N acquisition, partitioning of ¹⁵N and NUE. Restriction of N supply until V8, and from V8 to physiological maturity significantly reduced grain yield and N-uptake in all hybrids. Irrespective of the level of N-deficiency in plant and timing when the labeled fertilizer was applied, the amount of ¹⁵N recovered in the matured plant was the same in all N treatments. It has been evident that maize continued to take up N beyond 3 weeks after silking and the later N was applied during the development, the higher proportion of it was partitioned to grains. Of the total ¹⁵N uptake, 78% was partitioned to kernels in the N₄ treatment compared to only 61% in the control. Our data showed no evidence of differential N uptake, remobilization and NUE in the SG or Leafy hybrids tested, but the timing of N application and level of N-deficiency in plant significantly influenced N uptake, remobilization and N-dynamics in maize.     

Amino acid content in xylem sap of regrowing alfalfa (Medicago sativa L.): Relations with N uptake,N2 fixation and N remobilization

During vegetative regrowth of Medicago sativa L., soil N, symbiotically fixed N₂ and N reserves meet the nitrogen requirements for shoot regrowth. Experiments with nodulated or non-nodulated plants were carried out to investigate the changes in N flows originating from the different N sources and in xylem transport of amino acids during regrowth. Exogenous N uptake, N₂ fixation and endogenous N remobilization were estimated by ¹⁵N labelling and amino acids in xylem sap were analysed. Removal of shoots resulted in great declines of exogenous N flows derived either from N₂ or from NH₄NO₃ during the first week of regrowth, thereafter recovery increased linearly. Mineral N uptake as well as N₂ fixation occurred mainly between the 10th and 18th day after removal of shoots while exogenous N assimilation in intact plants remained at a steady level. Nitrogen remobilization rates in defoliated plants increased by at least three to five-fold, especially during the first 10 days following shoot removal. Compared to control plants, contents of amino acids in xylem sap, during the first 10 days of regrowth, were reduced by about 72% and 82% in NH₄NO₃ grown and in N₂ fixing plants, respectively. Asparagine was the main amino acid transported in xylem sap of both treated plants. Its relative contents during this period significantly decreased from 75% to 59% and from 67% to 36% respectively in non-nodulated plants and in nodulated ones. This decline was accompanied by compensatory increase in the relative contents of aspartate and glutamine.     

Decorrelating source and sink determinism of nitrogen remobilization during grain filling in wheat

Background and Aims

Nitrogen (N) remobilization is the major source of N for grain filling in wheat, the other being N uptake after anthesis (N_up); however, variations in remobilization efficiency are not fully understood. It is hard to tell whether the source or the sink effects predominate, because N in the culm at anthesis (N_ant) correlates strongly with both N remobilization (N_rem) and grain number (G_n), respectively the main source and the main sink.

Methods

A pot experiment was thus designed to assess the relative contributions of the source and sink to N_rem regulation. Using two cultivars of winter wheat (Triticum aestivum, ‘Apache’ and ‘Autan’), three pre-anthesis and two post-anthesis N fertilization levels were applied in order to vary the N sources, while ear trimming at anthesis reduced sink size.

Key Results

Unlike results observed at a scale of m², the equation binding N_ant to N_rem exhibited a negative intercept, challenging the concept of nitrogen remobilization efficiency. Before ear trimming, G_n fitted well to N_ant, with a slope dependent on genotype. To obtain a sink variable that was less correlated with N_ant, the difference δG_n was calculated between actual grain number and that which could be predicted from culm N before trimming. A multiple regression then predicted N_rem (r² = 0·95) from N_ant, N_up and δG_n, with fitting unbiased by fertilization treatment, trimming or genotype.

Conclusions

In untrimmed culms, δG_n had a negligible effect, so that N_rem could be fitted to N_ant and N_up only: grain N filling appeared to be determined by sources only (N_ant and N_up), not by sink, and the reduction of N_rem by N_up was quantified. In these ‘normal’ cases, the regulation of N_rem should thus be located within the N sources themselves. In contrast, ear-trimming needs to be considered with caution as it introduced a sink limitation on N_rem; moreover one with an important genotype effect.Key words: Triticum aestivum, winter wheat, source/sink, grain filling, nitrogen uptake, grain number, nitrogen harvest index, nitrogen remobilization efficiency, genotype × environment     

N and C NMR determination of allantoin metabolism in developing soybean cotyledons

The metabolism of allantoin by immature cotyledons of soybean (Glycine max L. cv Elf) grown in culture was investigated using solid state ¹³C and ¹⁵N nuclear magnetic resonance. All of the nitrogens of allantoin were incorporated into protein in a manner similar to that of each other and to the amide nitrogen of glutamine. The C-2 of allantoin was not incorporated into cellular material; presumably it was lost as CO₂. About 50% of the C-5 of allantoin was incorporated into cellular material as a methylene carbon; the other 50% was presumably also lost as CO₂. The ¹³C-¹⁵N bonds of [5-¹³C;1-¹⁵N] and [2-¹³C;1,3-¹⁵N]allantoin were broken prior to the incorporation of the nitrogens into protein. These data are consistent with allantoin's degradation to two molecules of urea and one two-carbon fragment. Cotyledons grown on allantoin as a source of nitrogen accumulated 21% of the nitrogen of cotyledons grown on glutamine. Only 50% of the nitrogen of the degraded allantoin was incorporated into the cotyledon as organic nitrogen; the other 50% was recovered as NH₄⁺ in the media in which the cotyledons had been grown. The latter results suggests that the lower accumulation of nitrogen by cotyledons grown on allantoin was in part due to failure to assimilate NH₄⁺ produced from allantoin. The seed coats had a higher activity of glutamine synthetase and a higher rate of allantoin degradation than cotyledons indicating that seed coats play an important role in the assimilation and degradation of allantoin.     

Heat stress effects on sink activity of developing maize kernels grown in vitro

The objective of this study was to determine the effect of short-term (4 days) and long-term (8 days) heat stress (35°C) on sink activity of maize (Zea mays L.) kernels. Beginning at 3 days after pollination (DAP) kernels were grown in vitro at 25°C and 24 h later were transferred to 35°C for either 4 or 8 days. Each treatment had a control that was maintained continously at 25°C. Two experiments were designed to examine the uptake and distribution of ¹⁴C among hexoses, sucrose and starch in the pedicel placento-chalazal (pedicel/p-c). endosperm, and pericarp tissues of kernels exposed to heat stress for 4 or 8 days. Kernels cultured in vitro were placed in ¹⁴C-sucrose medium either during the period of heat stress (experiment 1; 5 to 13 DAP) or immediately following heat-stress treatments (experiment 2; 10 to 22 DAP). In both experiments no significant effect of heat stress was observed on the total radioactivity accumulated in the kernels until about 17 DAP, after which heat-stressed kernels accumulated less ¹⁴C than the control. During the linear fill period, the endosperm of kernels exposed to heat stress accumulated more radioactivity associated with hexoses and sucrose and less radioactivity incorporated into starch, as compared to the control. Kernels heat stressed for 4 days showed a partial recovery in starch synthesis by 21 DAP, but to levels of only 65% of that of the control. Kernels heat stressed for 8 days did not recover. When ¹⁴C-sucrose was supplied during the heat stress period (5–13 DAP). kernels from all treatments accumulated more hexoses that sucrose in the pedicel/p-c. However, during the period following heat stress (10–22 DAP), pedicel/p-c accumulated sucrose, but only in kernels exposed to long-term heat stress. Soluble invertase activity was inhibited by both short-term and long-term heat stress, whereas the activity of insoluble invertase was affected only by long-term heat stress. These results support the hypothesis that the disruption of kernel growth and more particularly endosperm starch biosynthesis, in response to heat stress, is mainly associated with changes in carbon utilization and partitioning between the different nonstructural carbohydrates within the endosperm rather than with a limitation in carbon supply to the kernel. Therefore, the effect on sink activity does not seem to be attributable to a thermal disruption of kernel uptake of sugars, but rather it is a consequence of heat perturbation of other physiological processes such as endosperm sugar metabolism and starch biosynthesis.     

Main stem sink manipulation in wheat : effects on nitrogen allocation to tillers

The role of main stem (MS) sink size on N use by field-grown soft red winter wheat (Triticum aestivum L. cv Hart) was determined. At Feeke's growth stage 8 (last leaf just visible), 100 micromoles of 99 atom% ¹⁵N-ammonium was injected into the lower MS. At anthesis, MS sink size was adjusted by removal of 0, 33, 66, and 100% of the MS spikelets; tiller spikes were left intact. The MS and tiller average kernel size was unaffected by MS sink manipulations. The MS kernel N concentration increased when MS spikelets were removed. Tiller kernel N concentrations were unaffected except when the entire MS reproductive sink was removed, which caused an increase in tiller kernel N concentration. Net losses of MS vegetative N during grain fill were similar for all treatments except for plants lacking MS spikelets, which mobilized 30% less N from the MS. Labeled N was predominately (>90%) associated with the insoluble reduced N fraction of plant tissues at anthesis. Allocation of labeled N to tillers was not proportional to reduction in MS sink size. These results indicate that the reproductive sink on an individual culm has first priority for vegetative N mobilized during grain fill even when sink demand is reduced substantially.     

Light Restriction Delays Leaf Senescence in Winter Oilseed Rape (Brassica napus L.)

Oilseed rape (Brassica napus L.) is a crop with a complex aerial architecture that can cause self-shading leading to a vertical light gradient over the foliage. Mutual shading between neighboring plants at a high sowing density also results in an alteration of photosynthetically active radiation (PAR) absorption by lower leaves. The aim of this study was to analyze the impact that light restriction on lower leaves has on shoot architecture, biomass production and allocation, nitrogen (N) fluxes, and progression of sequential senescence. Field-grown plants were collected at the end of the vegetative rest period and grown in hydroponic conditions until pod maturity. A shading treatment corresponding to a 43.4 % reduction of PAR was applied at the early flowering stage. N uptake and fluxes of N allocation and remobilization were determined by supplying K¹⁵NO₃ in the nutrient solution. Photosynthesis and expression of SAG12 and Cab genes (indicators of leaf senescence progression) were also analyzed on different leaf ranks. The results showed that shading enhanced leaf development on the main stem and ramifications to optimize light capture. The expression pattern of the SAG12/Cab molecular indicator suggested a delay in leaf senescence that allowed leaf life span to be extended resulting in a more efficient leaf compound remobilization, with lower N residual contents in fallen leaves under shading. N uptake increased and N remobilization fluxes were enhanced from source organs (leaves and stem) toward sink organs (flowers). Profuse branching and late senescing varieties would be of interest for further selection programs under high sowing densities.     

Robustness of central carbohydrate metabolism in developing maize kernels

The central carbohydrate metabolism provides the precursors for the syntheses of various storage products in seeds. While the underlying biochemical map is well established, little is known about the organization and flexibility of carbohydrate metabolic fluxes in the face of changing biosynthetic demands or other perturbations. This question was addressed in developing kernels of maize (Zea mays L.), a model system for the study of starch and sugar metabolism. ¹³C-labeling experiments were carried out with inbred lines, heterotic hybrids, and starch-deficient mutants that were selected to cover a wide range of performances and kernel phenotypes. In total, 46 labeling experiments were carried out using either [U-¹³C₆]glucose or [U-¹³C₁₂]sucrose and up to three stages of kernel development. Carbohydrate flux distributions were estimated based on glucose isotopologue abundances, which were determined in hydrolysates of starch by using quantitative ¹³C-NMR and GC-MS. Similar labeling patterns in all samples indicated robustness of carbohydrate fluxes in maize endosperm, and fluxes were rather stable in response to glucose or sucrose feeding and during development. A lack of ADP-glucose pyrophosphorylase in the bt2 and sh2 mutants triggered significantly increased hexose cycling. In contrast, other mutations with similar kernel phenotypes had no effect. Thus, the distribution of carbohydrate fluxes is stable and not determined by sink strength in maize kernels.     

Distribution of Abscisic Acid in Maize Kernel during Grain Filling

The distribution of abscisic acid (ABA) within maize (Zea mays L.) kernels was studied in kernels from nontreated plants, from plants in which assimilate supply had been altered by source/sink manipulations, and in kernels cultured in vitro on ABA-free media. Prior to growth of the embryo, both the pedicel/placento-chalazal complex and the endosperm contained high concentration of ABA; however, the quantity of ABA in these tissues declined as the concentration in the embryo increased during the early stages of embryo growth. Peaks in the levels of ABA appeared to occur prior to and not concurrent with physiological events during grain filling. During most of the grain filling period, ABA concentration in the embryo was higher than that found in other kernel components. Altering assimilate supply by partial defoliation at two stages of development resulted in variable and transient effects on the relative distribution and concentration of ABA in kernel components. The concentration and distribution of ABA among components of kernels grown in vitro was similar to that observed for field-grown kernels. On the basis of these findings, in situ synthesis of ABA by kernel components is implicated and the putative role of ABA in the regulation of kernel development is discussed.     

Structure and Biosynthesis of Cuticular Lipids: Hydroxylation of Palmitic Acid and Decarboxylation of C(28), C(30), and C(32) Acids in Vicia faba Flowers

The structure and composition of the cutin monomers from the flower petals of Vicia faba were determined by hydrogenolysis (LiAlH₄) or deuterolysis (LiAlD₄) followed by thin layer chromatography and combined gas-liquid chromatography and mass spectrometry. The major components were 10, 16-dihydroxyhexadecanoic acid (79.8%), 9, 16-dihydroxyhexadecanoic acid (4.2%), 16-hydroxyhexadecanoic acid (4.2%), 18-hydroxyoctadecanoic acid (1.6%), and hexadecanoic acid (2.4%). These results show that flower petal cutin is very similar to leaf cutin of V. faba. Developing petals readily incorporated exogenous [1-¹⁴C]palmitic acid into cutin. Direct conversion of the exogeneous acid into 16-hydroxyhexadecanoic acid, 10, 16-dihydroxy-, and 9, 16-dihydroxyhexadecanoic acid was demonstrated by radio gas-liquid chromatography of their chemical degradation products. About 1% of the exogenous [1-¹⁴C]palmitic acid was incorporated into C₂₇, C₂₉, and C₃₁n-alkanes, which were identified by combined gas-liquid chromatography and mass spectrometry as the major components of the hydrocarbons of V. faba flowers. The radioactivity distribution among these three alkanes (C₂₇, 15%; C₂₉, 48%; C₃₁, 38%) was similar to the per cent composition of the alkanes (C₂₇, 12%; C₂₉, 43%; C₃₁, 44%). [1-¹⁴C]Stearic acid was also incorporated into C₂₇, C₂₉, and C₃₁n-alkanes in good yield (3%). Trichloroacetate, which has been postulated to be an inhibitor of fatty acid elongation, inhibited the conversion of [1-¹⁴C]stearic acid to alkanes, and the inhibition was greatest for the longer alkanes. Developing flower petals also incorporated exogenous C₂₈, C₃₀, and C₃₂ acids into alkanes in 0.5% to 5% yields. [G-³H]n-octacosanoic acid (C₂₈) was incorporated into C₂₇, C₂₉, and C₃₁n-alkanes. [G-³H]n-triacontanoic acid (C₃₀) was incorporated mainly into C₂₉ and C₃₁ alkanes, whereas [9, 10, 11-³H]n-dotriacontanoic acid (C₃₂) was converted mainly to C₃₁ alkane. Trichloroacetate inhibited the conversion of the exogenous acids into alkanes with carbon chains longer than the exogenous acid, and at the same time increased the amount of the direct decarboxylation product formed. These results clearly demonstrate direct decarboxylation as well as elongation and decarboxylation of exogenous fatty acids, and thus constitute the most direct evidence thus far obtained for an elongation-decarboxylation mechanism for the biosynthesis of alkanes.     

Pollen source effects on growth of kernel structures and embryo chemical compounds in maize

Background and Aims

Previous studies have reported effects of pollen source on the oil concentration of maize (Zea mays) kernels through modifications to both the embryo/kernel ratio and embryo oil concentration. The present study expands upon previous analyses by addressing pollen source effects on the growth of kernel structures (i.e. pericarp, endosperm and embryo), allocation of embryo chemical constituents (i.e. oil, protein, starch and soluble sugars), and the anatomy and histology of the embryos.

Methods

Maize kernels with different oil concentration were obtained from pollinations with two parental genotypes of contrasting oil concentration. The dynamics of the growth of kernel structures and allocation of embryo chemical constituents were analysed during the post-flowering period. Mature kernels were dissected to study the anatomy (embryonic axis and scutellum) and histology [cell number and cell size of the scutellums, presence of sub-cellular structures in scutellum tissue (starch granules, oil and protein bodies)] of the embryos.

Key Results

Plants of all crosses exhibited a similar kernel number and kernel weight. Pollen source modified neither the growth period of kernel structures, nor pericarp growth rate. By contrast, pollen source determined a trade-off between embryo and endosperm growth rates, which impacted on the embryo/kernel ratio of mature kernels. Modifications to the embryo size were mediated by scutellum cell number. Pollen source also affected (P < 0·01) allocation of embryo chemical compounds. Negative correlations among embryo oil concentration and those of starch (r = 0·98, P < 0·01) and soluble sugars (r = 0·95, P < 0·05) were found. Coincidently, embryos with low oil concentration had an increased (P < 0·05–0·10) scutellum cell area occupied by starch granules and fewer oil bodies.

Conclusions

The effects of pollen source on both embryo/kernel ratio and allocation of embryo chemicals seems to be related to the early established sink strength (i.e. sink size and sink activity) of the embryos.Key words: Zea mays, maize, pollen, kernel, embryo, endosperm, oil, protein, starch, soluble sugars     

Dynamics of nitrogen remobilization in defoliated Phleum pratense and Festuca pratensis under short and long photoperiods

The impact of photoperiod on the rate and magnitude of N remobilization relative to uptake of inorganic N during the recovery of shoot growth after a severe defoliation was compared over 18 days in two temperate grass species, timothy (Phleum pratense L. cv. Bodin) and meadow fescue (Festuca pratensis Huds. cv. Salten). Plants were grown in flowing solution culture with N supplied as 20 mM NH₄NO₃ and pre-treated by extending the 11 h photosynthetically significant light period either by 1 h (short-day or SD plants) or 7 h (long-day or LD plants) of very low light intensity, during the 10 days prior to defoliation. Following a single severe defoliation, ¹⁵N-labelled NH₄⁺ or NH₄⁺+ NO₃^? was supplied over a 20-day recovery period under the same SD and LD conditions. Changes in the relative contributions of remobilized N and newly acquired mineral N to shoot regrowth were assessed by sequential harvests. Both absolute and relative rates of N remobilization from root and stubble fractions were higher in LD than SD plants of both species, with the enhancement more acute but of shorter duration in timothy than fescue. Remobilized N was the predominant source of N for shoot regrowth in all treatments between days 0 and 8 after cutting; on average more so for fescue than timothy, because the presence of NO₃^? reduced the proportional contribution of remobilized N to the regrowth of timothy but not of fescue. Net uptake of mineral N began to recover between days 4 and 6 after cutting, with NO₃^? uptake restarting 1 or 2 days earlier than NH₄⁺ uptake, even when NH₄⁺ was the only form of N supply. LD timothy plants supplied solely with NH₄⁺ were slowest to resume uptake of mineral N. Supplying NO₃^? in addition to NH₄⁺ after defoliation promoted shoot regrowth rate but not remobilization of N. Rates of regrowth (shoot dry weight production per plant) were not correlated with rates of N remobilization from stubble either over the short-term (days 0–8) or longer term (days 0–18), interpreted as evidence against a causal dependence of regrowth rate on N remobilization under these conditions. The results are discussed in relation to hypotheses for source/sink-driven rates of N remobilization and their interactions with mineral N uptake following defoliation.     

Fusarium species complex and mycotoxins in grain maize from maize hybrid trials and from grower's fields

Aims: To quantify and to compare the occurrence of Fusarium species in maize kernels and stalk pieces, to analyse mycotoxins in kernels and maize crop residues, to evaluate two approaches to obtain kernel samples and to compare two methods for mycotoxin analyses. Methods and Results: The occurrence of Fusarium species in maize kernels and stalk pieces from a three‐year maize hybrid trial and 12 kernel samples from grower’s fields was assessed. Nine to 16 different Fusarium species were detected in maize kernels and stalks. In kernels, F. graminearum, F. verticillioides and F. proliferatum were the most prevalent species whereas in stalks, they were F. equiseti, F. proliferatum and F. verticillioides. In 2006, 68% of the kernel samples exceeded the recommended limit for pig feed for deoxynivalenol (DON) and 42% for zearalenone (ZON), respectively. Similarly, 75% of the samples from grower’s fields exceeded the limits for DON and 50% for ZON. In maize crop residues, toxin concentrations ranged from 2·6 to 15·3 mg kg^?1 for DON and from 0·7 to 7·4 mg kg^?1 for ZON. Both approaches to obtain maize kernel samples were valid, and a strong correlation between mycotoxin analysis using ELISA and LC‐MS/MS was found. Conclusions: The contamination of maize kernels, stalk pieces and remaining crop residues with various mycotoxins could pose a risk not only to animal health but also to the environment. With the hand‐picked sample, the entire Fusarium complex can be estimated, whereas combine harvested samples are more representative for the mycotoxin contents in harvested goods. Significance and Impact of the Study: This is the first multi‐year study investigating mycotoxin contamination in maize kernels as well as in crop residues. The results indicate a high need to identify cropping factors influencing the infection of maize by Fusarium species to establish recommendations for growers.