Effect of nitrogen nutrition on endosperm protein synthesis in wild and cultivated barley grown in spike culture

In normal growth conditions, total protein percent (salt soluble plus hordein fractions) in the endosperm at maturity in barley cultivar Hordeum vulgare L. cv `Ruth' was about 14%, whereas in an accession of wild barley, Hordeum spontaneum Koch line 297, it was about 28%. Spike culture experiments were conducted to ascertain whether there were basic differences between the two genotypes under conditions of widely different nitrogen supply. Spikes of each genotype were grown from 8 to 25 days after flowering in in vitro culture in a growth medium containing 0 to 4 grams per liter nitrogen supplied as NH<sub>4</sub>NO<sub>3</sub>. Spikes were pulse-labeled at intervals from 12 to 24 days after flowering with 3.7 megabecquerel of [<sup>3</sup>H]leucine to determine relative rates of synthesis of hordein-1 and hordein-2 polypeptides. At low nitrogen levels `Ruth' had a lower protein content than 297, but at increasing nitrogen levels its protein content increased rapidly and reached a maximum (35%) higher than 297 (30%). The relative contribution of the hordein fraction to total protein increased mainly with time, and hordein-1 to total hordein increased mainly with nitrogen level, in both genotypes. There appeared to be no fundamental limitations in the capacity of `Ruth' to accumulate protein; 297 appears to have a greater basal level of nitrogen availability under normal conditions.     

Synthesis of salt-soluble proteins in barley. Pulse-labeling study of grain filling in liquid-cultured detached spikes

The accumulation of salt-soluble proteins in the endosperm of developing barley (Hordeum vulgare L.) grains was examined. Detached spikes of barley were cultured at different levels of nitrogen nutrition and pulse-labeled with [¹⁴C] sucrose at specific times after anthesis. Proteins were extracted from isolated endosperms and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and crossed immunoelectrophoresis. Fluorography revealed an early, middle and late synthesis of specific proteins during grain filling. Synthesis of proteins appearing at the later stages responded to increased nitrogen nutrition. Two major components, -amylase and protein Z in particular, had a synthesis profile almost identical to that of the endosperm storage protein, hordein.Abbreviations CIE Crossed immunoelectrophoresis - SDSPAGE Sodium dodecyl sulphate polyacrylamide gel electrophoresis     

A role for γ3 hordein in the transport and targeting of prolamin polypeptides to the vacuole of developing barley endosperm

Hordein synthesis, transport and deposition was analysed by immunocytochemistry in developing endosperm cells of wild-type (Carlsberg II) and mutant varieties deficient in B hordein ( hor2ca ), γ1 hordein (Donetsky), γ2 hordein and minor B hordein polypeptides (Haisa), or γ3 hordein (Nevsky). In all varieties, hordein polypeptides were detected both in the cytoplasm as globules, ranging in diameter from 50 nm to 1.24 µm, and in the vacuole as protein bodies. In the cytoplasmic globules B and C hordein polypeptides are assembled as a core and are surrounded by an outer layer of γ1 and γ2 hordein. The globules apparently fuse several times in the cytoplasm before entering the vacuole. Absence of γ3 hordein in the mutant Nevsky leads to a dramatic change in hordein polypeptide targeting, the hordein storage proteins being largely deposited in the lumen of the rough endoplasmic reticulum. γ3 Hordein is unique among the sulphur-rich hordein polypeptides, being monomeric and forming only intramolecular disulphide bridges, while the other B and γ hordein polypeptides are aggregated by intermolecular disulphide bridges. Retention of hordein in the rough endoplasmatic reticulum in the absence of γ3 hordein suggests that γ3 hordein may maintain the prolamin storage polypeptides in a transport competent state. The sequence of the mature γ3 hordein polypeptide was deduced from a cDNA clone, and compared with γ2 hordein. The epitope recognized by the γ1 + γ2 hordein-specific BX monoclonal antibody used for immunocytochemistry was mapped to include E¹⁹⁰ and K¹⁹³, by synthesizing overlapping oligopeptides.     

Storage Protein Synthesis during Oat (Avena sativa L.) Seed Development

Oat (Avena sativa L.) seeds harvested at 2-day intervals from anthesis to maturity were tested for their ability to incorporate [³⁵S]sulfate into protein. Incorporation of [³⁵S]sulfate into TCA-insoluble material began 2 to 4 days postanthesis (DPA), reached a peak 14 to 16 DPA, and was barely detectable by 24 DPA. Incorporation of label into globulin was parallel to total protein accumulation, and averaged about 85% of the total protein synthesis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total protein extracted from developing seeds indicated that some polypeptides coinciding with the α and β globulin subunits were present 2 to 4 DPA, but the full complement of globulin polypeptides was not present until 10 DPA. Immunoprecipitation of in vivo labeled seed extracts showed that globulin polypeptides and the 59 kilodalton precursor were present at early stages of development (4 DPA). Quantitation of dot blot analysis, using an oat globulin cDNA clone as a probe, indicated that one species of oat globulin mRNA was most abundant 15 DPA, which is during the peak time of storage protein synthesis.     

Temporal dynamics in wheat grain zinc distribution: is sink limitation the key?

Background and Aims

Enhancing the zinc (Zn) concentration in wheat (Triticum aestivum) grain is a breeding objective in order to improve human Zn nutrition. At enhanced plant Zn uptake, grain Zn levels do not increase proportionally and within the grain the endosperm Zn levels remain below grain Zn levels. This study analysed the temporal dynamics of Zn concentrations in grain tissues during grain filling to find major bottlenecks.

Methods

Plants of two cultivars were grown at 1 and 5 mg Zn kg⁻¹ soil. Individual panicles were harvested 7, 14, 24 or 34 d after their flowering or at maturity and seeds were dissected into constituting tissues, which were analysed for Zn and other minerals.

Key Results

The Zn concentration of the crease was found to increase five- to nine-fold between 7 and 34 d after anthesis, while that of the endosperm decreased by 7 and 45 % when grown at 1 or 5 mg Zn kg⁻¹, respectively. The Zn turnover rate (d⁻¹) in the crease tissues was either independent of the Zn application level or higher at the lower Zn application level, and the Zn concentration increased in the crease tissues with time during grain filling while the turnover rate gradually decreased.

Conclusions

There is significant within-seed control over Zn entering the seed endosperm. While the seed crease Zn concentration can be raised to very high levels by increasing external Zn supply, the endosperm Zn concentrations will not increase correspondingly. The limited transfer of Zn beyond the crease requires more research to provide further insight into the rate-determining processes and their location along the pathway from crease to the deeper endosperm     

Effects of timing and supply of nitrogen on nitrogen remobilization from vegetative organs and redistribution to developing seeds of sunflower

A glasshouse study was made of the distribution of ¹⁵N among vegetative organs of sunflower and its later remobilization and redistribution to seeds, as influenced by the developmental stage at which ¹⁵N was provided, and by the N status of the plants. Plants of Hysun 30 sunflower were grown in sand culture and provided with K¹⁵NO₃ for a 3-day period at: (a) 3 days before the end of floret initiation; (b) 3 days before anthesis; (c) the start of anthesis; (d) full anthesis; and (e) 8 days after full anthesis. The plants were grown on a range of N supply rates, from severely deficient to more than adequate for maximum growth. Nitrogen-15 was distributed to all parts of the plant at the end of the ¹⁵N uptake periods. With the exception of the most N-stressed plants, subsequent remobilization of ¹⁵N from roots, stems and leaves occurred irrespective of the time the ¹⁵N was taken up. However, the percentage redistribution to seeds of ¹⁵N taken up at the end of floret initiation was less than for ¹⁵N taken up at anthesis. Remobilization of ¹⁵N from leaves and roots was higher (70%) for ¹⁵N taken up during and after anthesis than for ¹⁵N taken up at the end of floret initiation (45%), except for plants grown on the lowest N supply. By contrast, remobilization of ¹⁵N from the stem was lower for ¹⁵N taken up after full anthesis (40%) than before or during anthesis (>70%). The proportion of ¹⁵N remobilized from the top third of the stem was less than that from the bottom third, and decreased with increasing plant N status. Nitrogen-15 taken up over the 3-day supply periods during anthesis contributed from 2 to 11% of the total seed N at maturity; the contribution to seeds was greatest for plants grown on the highest N supply. Nitrogen taken up just before and during anthesis contributed most of the N accumulated in mature seeds of plants grown on an adequate N supply, but N taken up between the end of floret initiation and just before anthesis, or after full anthesis seemed to make an equally important contribution to mature seeds as N taken up during anthesis for plants grown on a very low N supply. It was concluded that the development of florets and seeds of sunflower is supported by N taken up by the plant between the end of floret initiation and anthesis, and by N redistributed from vegetative organs. Unless soil N is so low as to impair early growth, split applications of N fertilizer would be best made just before the end of floret initiation (‘star stage’) and just before anthesis.     

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.     

Protein Synthesis during the Initial Phase of the Temperature-Induced Bleaching Response in Euglena gracilis

Growing cultures of photoheterotrophic Euglena gracilis experience an increase in chlorophyll accumulation during the initial phase of the temperature-induced bleaching response suggesting an increase in the synthesis of plastid components at the bleaching temperature of 33°C. A primary goal of this work was to establish whether an increase in the synthesis of plastid proteins accompanies the observed increase in chlorophyll accumulation. In vivo pulse-labeling experiments with [³⁵S]sodium sulfate were carried out with cells grown at room temperature or at 33°C. The synthesis of a number of plastid polypeptides of nucleocytoplasmic origin, including some presumably novel polypeptides, increased in cultures treated for 15 hours at 33°C. In contrast, while synthesis of thylakoid proteins by the plastid protein synthesis machinery decreased modestly, synthesis of the large subunit of the enzyme ribulosebisphosphate carboxylase was strongly affected at the elevated temperature. Synthesis of novel plastid-encoded polypeptides was not induced at the bleaching temperature. It is concluded that protein synthesis in plastids declines during the initial phase of the temperature response in Euglena despite an overall increase in cellular protein synthesis and an increase in chlorophyll accumulation per cell.     

Evolutionary relationship of the members of the sulphur-rich hordein family revealed by common antigenic determinants

Summary Five monoclonal antibodies raised against an enriched C hordein fraction have been characterized in detail and were found to be specific for the members of the sulphur-rich hordein family. Two antibodies specific for B hordein polypeptides were identified, one of which reacted predominantly with CNBr cleavage class III polypeptides. 1 hordein was recognized by two antibodies, of which one also reacted with 2 hordein and several members of the CNBr cleavage class II B hordein polypeptides. One antibody recognized 3 hordein but cross-reacted at higher antibody concentration with almost all of the B and C hordein polypeptides. The specificity of the monoclonal antibodies was confirmed by Western blotting of one- or two-dimensionally separated hordein from the B hordein-deficient mutant hor2ca and its wild-type Carlsberg II and the 3 hordein-deficient genotype Nevsky. The identification of the hordein-specific monoclonal antibodies was further supported by immune precipitation of in-vitro transcribed and translated 2 hordein, and hor2ca and Carlsberg II mRNA translation products. The monoclonal antibodies were used to screen for mutants in hordein synthesis. Two mutants, one deficient in 1 hordein synthesis and a second in 2 or closely related B hordein polypeptides were identified. A model is proposed for the evolution of the sulphur-rich hordein loci Hor5 and Hor2.     

氮肥运筹对晚播冬小麦氮素和干物质积累与转运的影响

氮素平衡对干物质积累与分配的影响是农业生态系统研究的重要内容,在保障产量前提下减少氮肥施用量可减少环境污染与温室气体排放。以晚播冬小麦为研究对象,设置4个施氮量水平:0 kg/hm2(N0)、168.75 kg/hm2(N1)、225 kg/hm2(N2)、281.25 kg/hm2(N3),每个施氮量水平下设置2个追氮时期处理:拔节期(S1)、拔节期+开花期(S2),研究了氮肥运筹对晚播冬小麦氮素和干物质积累与转运及氮肥利用率的影响。结果表明:拔节期追施氮肥(S1)条件下,在225 kg/hm2(N2)基础上增施25%氮肥(N3)对开花期氮素积累总量和营养器官氮素转运量无显著影响;拔节期+开花期追施氮肥(S2)条件下,随施氮量增加,开花期氮素积累总量和花后营养器官氮素转运量升高;S2较S1显著提高成熟期籽粒及营养器官氮素积累量、花后籽粒氮素积累量及其对籽粒氮素积累的贡献率。同一施氮量条件下,S2较S1提高了成熟期的干物质积累量、开花至成熟阶段干物质积累强度和花后籽粒干物质积累量。同一追氮时期条件下,籽粒产量N2与N3无显著差异,氮肥偏生产力随施氮量增加而降低;同一施氮量条件下,S2较S1提高了晚播冬小麦的籽粒产量和氮肥吸收利用率。拔节期+开花期追施氮肥,总施氮量225kg/hm2为有利于实现晚播冬小麦高产和高效的最优氮肥运筹模式。     

Understanding Dry Matter and Nitrogen Accumulation with Time-Course for High-Yielding Wheat Production in China

Understanding the time-course of dry matter (DM) and nitrogen (N) accumulation in terms of yield–trait relationships is essential to simultaneously increase grain yield and synchronize N demand and N supply. We collected 413 data points from 11 field experiments to address patterns of DM and N accumulation with time in relation to grain yield and management of winter wheat in China. Detailed growth analysis was conducted at the Zadok growth stages (GS) 25 (regreening), GS30 (stem elongation), GS60 (anthesis), and GS100 (maturity) in all experiments, including DM and N accumulation. Grain yield averaged 7.3 Mg ha⁻¹, ranging from 2.1 to 11.2 Mg ha⁻¹. The percent N accumulation was consistent prior to DM accumulation, while both DM and N accumulation increased continuously with growing time. Both the highest and fastest DM and N accumulations were observed from stem elongation to the anthesis stage. Significant correlations between grain yield and DM and N accumulation were found at each of the four growth stages, although no positive relationship was observed between grain yield and harvest index or N harvest index. The yield increase from 7–9 Mg ha⁻¹ to >9 Mg ha⁻¹ was mainly attributed to increased DM and N accumulation from stem elongation to anthesis. Although applying more N fertilizer increased N accumulation during this stage, DM accumulation was not improved, indicating that N fertilizer management and related agronomic management should be intensified synchronously across the wheat growing season to simultaneously achieve high yields and match N demand and N supply.     

Assessing post-anthesis nitrogen uptake, distribution and utilisation in grain protein synthesis in barley (Hordeum vulgare L.) using 15N fertiliser and 15N proteinogenic and non-proteinogenic amino acids

We investigated the effects of nitrogen (N) availability during the vegetative phase on (a) post‐anthesis N uptake and (b) its translocation into ears in barley plants grown in a greenhouse at two levels of N: low (50 mg N kg^?1 sand) and optimal N supply (150 mg N kg^?1 sand). Plants in the two N treatments were fertilised with the same amount of labelled ¹⁵N [50 mg ¹⁵N kg^?1 sand at 10% ¹⁵N_exc (N_excess, i.e. N_exc, is defined as the abundance of enriched stable isotope minus the natural abundance of the isotope) applied as ¹⁵NH₄¹⁵NO₃] 10 days after anthesis (daa). In a separate experiment, the uptake and transport into ears of proteinogenic and non‐proteinogenic amino acids were studied to determine whether a relationship exists between amino acid transport into ears and their proteinogenic nature. Plants were fed with either ¹⁵N‐α‐alanine, a proteinogenic amino acid, or ¹⁵N‐α‐aminoisobutyric acid, a non‐proteinogenic amino acid. Both these amino acids were labelled at 95.6% ¹⁵N_exc. Results showed that N accumulations in stems, leaves and especially in ears were correlated with their dry matter (dm) weights. The application of 150 mg N kg^?1 sand significantly increased plant dm weight and total N accumulation in plants. During their filling period, ears absorbed N from both external (growth substrate) and internal (stored N in plants) sources. Nitrogen concentration in ears was higher in optimal N‐fed plants than in low N‐fed plants until 10 daa, but from 21 to 35 daa, differences were not detected. Conversely, ¹⁵N_exc in ears, leaves and stems was higher in low N‐fed plants than in optimal N‐fed plants. Ears acted as strong sink organ for the post‐anthesis N taken up from the soil independently of pre‐anthesis N nutrition: on average, 87% of the N taken up from the soil after anthesis was translocated and accumulated in ears. Low N‐fed plants continued to take up N from the post‐anthesis N fertiliser during the later grain‐filling period. The increase of pre‐anthesis N supply rate led to a decrease in the contribution of nitrogen derived from post‐anthesis ¹⁵N‐labelled fertiliser (N_dff) to total N in all aboveground organs, especially in ears where 44% and 22% of total N originated from post‐anthesis N uptake in low N‐fed and optimal N‐fed plants, respectively. The experiment with labelled amino acids showed that there was greater transport of proteinogenic amino acid into the ear (50% of total ¹⁵N) than non‐proteinogenic amino acid (39%). However, this transport of the non‐proteinogenic amino acids into ear suggested that the transport of N compounds from source (leaves) to sink organs (ear) might not be intrinsically regulated by their ability to be incorporated into storage protein of ears.     

The Extraction, Solubility, and Characterization of Two Groups of Barley Storage Polypeptides

This paper reports further studies on the characteristics ofthe storage protein fraction (hordein) of barley. Hordein consistsof two groups of polypeptides (termed ‘B’ and ‘C’)coded by two separate but linked loci. Whereas the ‘C’polypeptides are readily soluble and extracted in 60% (v/v)ethanol at room temperature, the ‘B’ group is moresoluble in, and therefore more efficiently extracted by, 50%(v/v) propan-1-ol or 45% (v/v) propan-2-ol at elevated temperaturesand in the presence of 2-mercaptoethanol. However, the mostefficient conditions for hordein extraction (50% propan-1-ol+ 2% (v/v) 2-mercaptoethanol at 60 °C) also extract somecontaminating non-hordein polypeptides resulting in an apparentlyincreased lysine content of the hordein fraction. Amino acid analysis of the purified ‘B’ and ‘C’hordein groups shows that, whereas ‘C’ hordein containsmore glutamate + glutamine, proline, and phenylalanine than‘B’ hordein, it contains only traces of lysine andsulphur amino acids in contrast to ‘B’ hordein whichcontains 0·5% lysine 0·6% methionine, and 2·5%cysteine. Equilibrium sedimentation analyses carried out on the purified‘B’ and ‘C’ groups indicates that thepreparations were reasonably monodisperse with molecular weightsof approximately 32 000 and 52 000 respectively. These valuesare considerably lower than those previously determined by SDS-PAGE.     

Phase-Specific Polypeptides and Poly(A) RNAs during the Cell Cycle in Synchronous Cultures of Catharanthus roseus Cells

This study shows an overall analysis of gene expression during the cell cycle in synchronous suspension cultures of Catharanthus roseus cells. First, the cellular cytoplasmic proteins were fractionated by two-dimensional gel electrophoresis and visualized by staining with silver. Seventeen polypeptides showed qualitative or quantitative changes during the cell cycle. Second, the rates of synthesis of cytoplasmic proteins were also investigated by autoradiography by labeling cells with [³⁵S]methionine at each phase of the cell cycle. The rates of synthesis of 13 polypeptides were found to vary during the cell cycle. The silverstained electrophoretic pattern of proteins in the G₂ phase in particular showed characteristic changes in levels of polypeptides, while the rates of synthesis of polypeptides synthesized during the G₂ phase did not show such phase-specific changes. This result suggests that posttranslational processing of polypeptides occurs during or prior to the G₂ phase. In the G₁ and S phases and during cytokinesis, several other polypeptides were specifically synthesized. Finally, the variation of mRNAs was analyzed from the autoradiograms of in vitro translation products of poly(A)⁺ RNA isolated at each phase. Three poly(A)⁺ RNAs increased in amount from the G₁ to the S phase and one poly (A)⁺ RNA increased preferentially from the G₂ phase to cytokinesis.     

Differential protein synthesis in response to sulphate and phosphate deprivation: Identification of possible components of plasma-membrane transport systems in cultured tomato roots

Isolated roots of Lycopersicon esculentum Mill., cultured in axenic conditions were starved of sulphate or phosphate, and uptake capacities for the respective oxyanion-transport systems were observed for several days after sulphate or phosphate withdrawal. Sulphate-uptake capacity of the intact roots, measured in a 20-min period, increased from a control level of 100 nmol · g^–1 · h^–1 to 1100 nmol · g^–1 · h^–1 in 10 d, and phosphate-uptake capacity increased from 500 to 1400 nmol · g^–1 · h^–1 over 4 d. Newly synthesised polypeptides of these root cultures were pulse-labelled in vivo for 2 h, by adding [³H]leucine to the culture medium. The tissue was immediately homogenised and soluble and membrane fractions were prepared. A highly purified plasma-membrane fraction was separated from the crude microsomal membrane fraction using an aqueous two-phase partitioning technique. All fractions were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and autoradiography. A 28-kilodalton (kDa) soluble polypeptide, and 36-, 43-, and 47-kDa plasma-membrane polypeptides were observed to have increased labelling after 4 d of sulphate deprivation. Longer periods resulted in additional polypeptides with increased [³H]leucine incorporation. The synthesis of a 25-kDa membrane polypeptide and a 65-kDa soluble polypeptide was increased after 4 d of phosphate deprivation. Two-dimensional electrophoresis afforded greater resolution of the plasmamembrane polypeptides, confirming increased synthesis of the 36-kDa polypeptide and the presence of the 28-kDa polypeptide in the plasma-membrane preparation from sulphate-starved roots. These polypeptides were also observed in protein-stained two-dimensional gels as low-abundant protein components of the plasmamembrane fraction. It is suggested that the 36-kDa polypeptide may be a component of the plasma-membrane sulphate-transport system and that the 25-kDa polypeptide may be a component of a phosphate-transport system.Abbreviations kDa kilodalton(s) - PAGE polyacrylamide gel electrophoresis - pI isoelectric point - SDS Sodium dodecyl sulphate This work was supported by the Agricultural and Food Research Council via grants-in-aid to Long Ashton Research Station. We are also grateful for discussions with our colleagues D.T. Clarkson (LARS) and J.-C. Davidian (ENSA/INRA, Montpellier).     

Metabolism of free sugars in relation to starch synthesis in the developing Sorghum vulgare grain

Accumulation of starch at expense of its free-sugar precursors was studied in the developing grains of the ‘SL-44’variety of Sorghum vulgare Pers. The content of starch gradually increased with the maturation of the grain and this increase was relatively fast until 18 days after anthesis. The daily rate of starch accumulation was at a maximum 15 days after anthesis. The content of total free sugars, reducing sugars, non-reducing sugars other than sucrose, total and non-sucrosyl fructose, and glucose also increased, reaching maximum values at 18 days after anthesis. Sucrose content gradually increased with a concomitant decrease in the activity of invertase, and sucrose was the major non-reducing sugar in the matured grains. Detached heads incubated in labelled sugars indicated that, compared to sucrose and fructose. ¹⁴C was more efficiently incorporated from glucose into grain starch, which was maximally synthesized at the mid-milky stage of grain development. Exogenous supply of NAD⁺ plus ATP stimulated the in vivo incorporation of ¹⁴C from sucrose to starch. The decline in the rate of starch accumulation did not synchronise with that of protein synthesis.     

Effects of variation in temperature and light intensity at different times on growth and yield of spring wheat

Spring wheat grown in pots outdoors was transferred to growth rooms for various periods to study the effect of increasing the temperature from 14-4 to 20-3 ^oC (Expt 1) or from day/night values of 15-0/15-2 to 20-0/15-2 ^oC (Expt 2) and of increasing the amount of visible radiation in a 16 h day from 424 to 792 J cm^-2 (Expt 1) or 374 to 740 J cm^-2 (Expt 2). There were no interactions between temperature and radiation. In Expt 1 neither the increase in temperature nor extra radiation, applied for 14 days immediately after the appearance of double ridges on the stem apex, or 14 days later, increased grain yield at maturity. Warmth early, but not late, increased dry weight, leaf area and the number of floret primordia immediately after treatment, but these effects had disappeared by anthesis, 30 days later. Dry weight but not leaf area was increased by extra radiation but the effects had disappeared 2 wk after treatment. An increase in temperature imposed for 16 days starting 5 or 21 days after anthesis (Expt 2) increased dry weight of the ear and decreased that of the rest of the plant immediately after treatment, and decreased leaf area at all times. When plants from the two temperatures were put together in the same conditions, ear growth of plants that had been in the warm was slower than that of plants from the cold treatment, so that the difference in ear weight observed after 16 days of treatment reversed and grain yield was decreased by warmth applied in either period; the component of yield decreased by warmth was grain size. Additional radiation in either post-anthesis period increased dry weight of all parts of the plant and had negligible effects on leaf area. Final grain yield was increased by c. 15% because the individual grains were larger. Early treatment also increased grain number slightly. The effects of treatment during the two post-anthesis periods were similar in size, and additive.     

Legume defoliation affects rhizosphere decomposers, but not the uptake of organic matter N by a neighbouring grass

Legume–grass interactions have a great influence on grassland primary production and it was recently shown how defoliation of a legume can increase the transfer of fixed N to a neighbouring grass. It has also been shown that defoliation of a plant can increase soil microbial activity and lead to better soil N availability in the rhizosphere of the defoliated plant. We combined these two perspectives and tested whether defoliation of a legume (Lotus corniculatus) can enhance N nutrition of the neighbouring grass (Holcus lanatus) by increasing growth of soil decomposer biota and the availability of soil organic matter N for grass uptake. We grew mixtures of L. corniculatus and H. lanatus in grassland soil that included ¹⁵N-labelled L. corniculatus litter. In half of the systems, we subjected L. corniculatus to a defoliation treatment mimicking insect larvae feeding. At destructive harvests 1, 3, 9 and 30 days after the last defoliation event, we determined how L. corniculatus defoliation affected decomposer microbes, protozoa and nematodes and whether these changes among decomposers created a feedback on the growth and ¹⁵N uptake of the neighbouring H. lanatus. Defoliation reduced the growth and litter-N uptake, but increased shoot N concentration of L. corniculatus. Of the soil variables measured, defoliation doubled the number of bacterial-feeding protozoa, but did not affect the abundance of decomposer microbes and bacterial- and fungal-feeding nematodes. Defoliation did not have statistically significant effects on H. lanatus shoot growth, shoot N concentration or litter-N uptake. Our results demonstrate how defoliation-induced changes in legume ecophysiology can affect the growth of decomposers in soil. However, these effects did not appear to lead to a significant change in the availability of soil organic N to the neighbouring grass. It seems that when positive effects of legume defoliation on grass N nutrition are found in grassland ecosystems, these are more likely to be explained by direct transfer of fixed N rather than changes in the availability of soil organic matter N.