Nitrogen transfer from nodulating soybean to maize or to nonnodulating soybean in intercrops: the 15N dilution method

In 1985, 1986 and 1988, maize (Zea mays L.) was monocropped or intercropped with nodulating or nonnodulating soybean (Glycine max [L.] Merr.). In addition, nodulating soybean and nonnodulating soybean were each monocropped and grown as a mixture. In 1985 and 1986, treatments were grown at 0 and 60 kg N ha^–1 and in 1988, the treatments were grown without N fertilizer, on N-depeted soil and on non-N-depleted soil. ¹⁵N enriched N was applied to soil in all the aforementioned treatments to test for N transfer from nodulating soybean to non-N₂-fixing crops by the ¹⁵N dilution method.The ¹⁵N dilution method did not show the occurrence of N transfer in 1985 and 1986, but the N sparing effect was evident from the total N uptake of nonnodulating soybean, dwarf maize and tall maize, in 1986. In 1988, maize and nonnodulating soybean seed yields and seed N yields were higher on non-N-depleted soil than on N-depleted soil. On N-depleted soil, the ¹⁵N dilution method indicated N transfer from nodulating soybean to maize and to nonndulating soybean. At a population ratio of 67% nodulating soybean to 33% nonnodulating soybean, N transfer was also seen on non-N-depleted soil in 1988.     

Nitrogen accumulation in sole and mixed stands of sweet-blue lupin (Lupinus angustifolius L.), ryegrass and oats

Nitrogen fixation was measured in monocropped sweet-blue lupin (Lupinus angustifolius), lupin intercropped with two ryegrass (Lolium multiflorum) cultivars or with oats (Avena sativa) on an Andosol soil, using the ¹⁵N isotope dilution method. At 117 days after planting and at a mean temperature below 10°C, monocropped lupin derived an average of 92% or 195 kg N ha⁻¹ of its N from N₂ fixation. Intercropping lupin with cereals increased (p<0.05) the percentage of N derived from atmospheric N₂ (% Ndfa) to a mean of 96%. Compared to the monocropped, total N fixed per hectare in intercropped lupin declined approximately 50%, in line with the decrease in seeding rate and dry matter yield. With these high values of N₂ fixation, selection of the reference crop was not a problem; all the cereals, intercropped or grown singly produced similar estimates of N₂ fixed in lupin. It was deduced from the ¹⁵N data that significant N transfer occurred from lupin to intercropped Italian ryegrass but not to intercropped Westerwoldian ryegrass or to oats. Doubling the ¹⁵N fertilizer rate from 30 to 60 kg N ha⁻¹ decreased % Ndfa to 86% (p<0.05), but total N fixed was unaltered. These results indicate that lupin has a high potential for N₂ fixation at low temperatures, and can maintain higher rates of N₂ fixation in soils of high N than many other forage and pasture legumes.     

Rate of differentiation and emergence of nodal maize roots

The timing of root production is one of the parameters required for modelling the root system architecture. The objectives of this study are (1) to describe the rate of appearance of adventitious root primordia of maize and their rate of emergence out of the stem; (2) to test equations for the prediction of the rank of the phytomer on which root emergence occurs, in a wide range of field situations.Maize, cultivar Dea, was grown in controlled conditions and in the field in 1987, 1988, 1989 and 1991. Plants were regularly sampled and the following data were recorded: foliar stage, number of root primordia and number of emerged roots per phytomer. Root primordia were counted in transverse thin sections in the stem.At a single plant level, root primordia differentiation occurred sequentially on the successive phytomers, with no overlapping between two phytomers. The same was true for root emergence. Roots belonging to the same phytomer emerged at approximately the same time.At a plant population level, there was a linear relationship between the rank of the phytomer on which root primordia were differentiated and cumulated degree-days after sowing. A linear relationship was also observed between the rank of the phytomer on which roots were emerging and cumulated degree-days or foliar stage. In the range of field situations tested (several years, sowing dates and planting densities), both equations gave an accurate prediction of the timing of root emergence during the plant cycle.     

Tissue-specific subcellular immunolocalization of a myosin-like protein in maize root apices

Summary Using a heterologous myosin antibody raised against the whole molecule of bovine muscle myosin, we have identified a myosin-like protein in maize. Immunoblots of subcellular fractions isolated from roots identified one distinct band at about 210 kDa in the microsomal protein fraction and one band at about 180 kDa in the soluble protein fraction. Indirect immunofluorescence was performed using maize root apex sections to reveal endocellular distributions of the myosin-like protein. Both diffuse and particulate labelling patterns were observed throughout the cytoplasm of all root cells. In mitotic cells, myosin-like protein was excluded from spindle regions. Amyloplast surfaces were labelled prominently in cells of the root cap statenchyma and in all root cortex cells. On the other hand, myosin-like protein was prominently enriched at cellular peripheries in cells of the pericycle and outer stele in the form of continuous peripheral labelling. From all root apex tissues, phloem elements showed the most abundant presence of myosinlike protein.Abbreviations AFs actin filaments - MTs microtubules Dedicated to Professor Walter Gustav Url on the occasion of his 70th birthday     

Reversion of Texas male-sterile cytoplasm maize in culture to give fertile,T-toxin resistant plants

Summary Plants carrying Texas male-sterile (Tms) cytoplasm are normally sensitive to Drechslera maydis T-toxin. Tissue cultures were initiated from immature embryos of maize carrying Tms-cytoplasm, and plants were regenerated after selection for resistance to T-toxin. Fertile, T-toxin resistant plants were obtained from the unselected control cultures as well as from the selected material. In addition, one regenerant from an unselected culture was fertile and T-toxin sensitive. The progeny of the regenerants showed the phenotype of the female parent with respect to pollen-fertility, and T-toxin resistance. The data are consistent with the heritable changes observed being the result of the expression of an altered mitochondrial genome.     

Determination of lead in white lupin by anodic stripping voltammetry

An anodic stripping voltammetry method for the simultaneous determination of trace heavy metals in nutrient solutions, soils and plants has been developed at a hydrodynamic electrochemical sensor. Several parameters were optimized in order to enhance sensitivity. Calibration curves in different media are presented. The study of the uptake of lead by white lupin (Lupinus albus L.) was carried out. Toxicity symptoms were observed and compared with levels of lead measured in roots and leaves.     

Depletion of macro-nutrients from rhizosphere soil solution by juvenile corn,cottonwood, and switchgrass plants

In situ sampling of rhizosphere solution chemistry is an important step in improving our understanding of soil solution nutrient dynamics. Improved understanding will enhance our ability to model nutrient dynamics and on a broader scale, to develop effective buffers to minimize nutrient movement to surface waters. However, only limited attention has been focused on the spatial heterogeneity and temporal dynamics of rhizosphere solution, and still less is known about how rhizosphere solution chemistry varies among plant species. Nutrients in rhizosphere soil solution and changes in root morphology of juvenile corn (Zea mays L. cv. Stine 2250), cottonwood (Populus deltoids L.), and switchgrass (Panicum virgatum L.) were monitored using mini-rhizotron technology. Plants were grown for 10 days in a fine-silty, mixed, superactive, mesic Cumulic Hapludoll (Kennebec series). Micro-samples (100–200 μL) of rhizosphere and bulk soil solution were collected at 24-h intervals at a tension of −100 kPa and analyzed for P, K, Ca, and Mg concentration using Capillary Electrophoresis techniques. Plants were harvested at the end of the 10-day period, and tissue digests analyzed for nutrient content by Inductively Coupled Plasma Spectroscopy. Corn plants produced roots that were 1.3 times longer than those of cottonwood, and 11.7 times longer than those of switchgrass. Similar trends were observed in number of root tips and root surface area. At the end of 10 days, rhizosphere solution P and K concentrations in the immediate vicinity of the roots (<1 mm) decreased by approximating 24 and 8% for corn, and 15 and 5% for cottonwood. A rhizosphere effect was not found for switchgrass. After correction for initial plant nutrient content, corn shoot P, K, and Mg were respectively 385, 132, and 163% higher than cottonwood and 66, 37, and 10% higher than switchgrass. Cottonwood shoot Ca concentration, however, was 68 to 133% higher than that of corn or switchgrass. There was no difference in root P concentration among the three species. Nutrient accumulation efficiency (μg nutrient mm⁻¹ root length) of cottonwood was 26 to 242% higher for P, 25 to 325% higher for Ca, and 41 to 253% higher for Mg than those of corn and switchgrass. However, K accumulation efficiency of corn was four to five times higher than that of the cottonwood and switchgrass. Nutrient utilization efficiency (mg of dry weight produced per mg nutrient uptake) of P, K, and Mg was higher in cottonwood than in corn and switchgrass. These differences are element-specific and depend on root production and morphology as well as plant nutrient status. From a practical perspective, the results of this study indicate that potentially significant differences in rhizosphere solution chemistry can develop quickly. Results also indicate that cottonwood would be an effective species to slow the loss of nutrients in buffer settings. An erratum to this article can be found at     

Quantitative estimation of root exudation of maize plants

Summary The rate of root exudation of maize plants was estimated by measuring the rate of denitrification in a hermetically sealed root system. While CO₂ production measured in the rhizosphere results both from root respiration and microbial respiration N₂O production during nitrate respiration is solely related to the amount of root exudates available for bacterial degradation. With 4 week old plants growing in quartz sand or soil root exudation amounted to 7% of the net photosynthates. Calculations revealed that about 25% of the organic matter flowing into the root system was excreted into the rhizosphere.     

The effect of oxygen deficiency on uptake and distribution of nutrients in maize plants

Young maize (Zea mays L.) plants, 7 days after germination were exposed to nutrient solutions which were either aerated or not aerated for 14 days. Nutrients were supplied as 50% strength Hoagland’s solution or, in the case of the four ‘low nutrient’ treatments, N, P, K or Ca were supplied at the equivalent of 10% strength Hoagland’s solution. Shoot fresh weight was decreased by 25% due to lack of aeration; O₂ deficiency also impaired leaf elongation but not dry weights, suggesting that lack of O₂ in the roots impaired cell expansion in shoots more than dry weight accumulation. The distribution of N, P, K and Ca within shoots was consistent with their relative mobilities in the phloem; at least 7% of Ca in plants after 14 days of treatments was found in the oldest leaf whereas N, P and K were rapidly remobilised to younger tissues. Between 33 and 49% of the total N, P and K in the shoot was found in the 40 mm of tissue at the base of the growing leaves in plants grown for 14 days at low nutrient concentrations. Concentrations (dry weight basis) of phloem-mobile nutrients were also greatest in the growing zones of the leaves, especially in the case of N and P. Calcium, on the other hand, was found in relatively low concentrations in the youngest tissue and as with the other nutrients, concentrations declined due to low external supply, non-aeration or a combination of both. In spite of the failure of Ca to move from old to young leaves, the effect of the deficiencies of N, P and K was probably as severe as that of Ca in the youngest tissues of treated plants. Calcium uptake by the whole shoot appeared to be slightly less sensitive to O₂ deficits than that of N, P and K. This compensated for the failure of Ca to move to growing tissues during periods of low external Ca supply.     

Production of transgenic maize plants and progeny by bombardment of hi-II immature embryos

Summary Production of transgenic maize (Zea mays L.) callus, plants, and progeny from microprojectile bombardment of 2–5-d cultured Hi-II immature embryos is described. Histological evidence indicates that these tissues are amenable to transformation due to surface layer cell division of the scutellum. Two out of every 100 bombarded embryos produced transgenic callus and R₀ transgenic plants were both male and female fertile. Expected segregation of transgenes was observed in progeny. The primary advantage of bombarding these tissues is increased male and female fertility of transgenic plants compared with those produced using long-term callus or suspension cultures.     

Tillage and N fertilization effects on maize root growth and root:shoot ratio

Two methods for estimating the size of the maize (Zea mays l.) root system from soil cores taken in the field were compared. The spatially weighed block method of estimation accounted for variation in root density by using 18 samples per plant which varied in distance from plant and soil depth. This method was compared to an estimation which averaged all of the 18 samples together. Both methods gave surprisingly similar estimates for total root growth. Increased root growth in the surface soil layers, due to tillage and N fertilization, did not impact on the estimation of total root growth. Total root length remained unchanged or increased with N fertilization, while root weight remained the same or decreased. Root mass per length decreased with N fertilization. The estimated size of the root system was used to calculate root:shoot weight ratios. The largest root:shoot ratio was found in the vegetative stage and decreased throughout the rest of the season. In this field experiment, the estimated size of the root system at 8 weeks after planting was not significantly different from the size at silking or harvest. Nitrogen fertilization significantly decreased the root:shoot weight ratio. However, tillage did not significantly change the ratio.     

The maximum axial growth pressure of roots of spring and autumn cultivars of lupin

The maximum axial growth pressure of roots of Lupinus albus cv Lublanc and Lupinus albus cv Lunoble (spring and autumn-sown cultivars respectively) were measured. The mean values were not significantly different with an overall mean value of 645 kPa. This value is not unusually large for plant roots and is surprising because lupins are known for their ability to penetrate strong soils. The autumn cultivar had a significantly greater maximum elongation rate under zero mechanical impedance than the spring cultivar. The impeded diameters were also larger in the autumn cultivars.     

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.     

Phloem translocation of gibberellins in three species of higher plants

Phloem sap was collected from white lupin (Lupinus albus L.), cowpea (Vigna unguiculata L.) and castor bean (Ricinus communis L.) and analysed for gibberellins (GAs) using gas chromatography-mass spectrometry (GC-MS). A large number of GAs were found in the phloem exudate of all three species, particularly where the sap was collected from pods (white lupin and cowpea) and in these legumes GAs representing both the early C-13-hydroxylation and non-hydroxylation pathways of biosynthesis were identified. In the sap collected from the vegetative tissues of castor bean the number of GAs identified was fewer than that in the other species, representing mainly the non-hydroxylation pathway. Data from sap collected from the pedicel and stylar ends of pods and by making feeds of radiolabelled GAs to seeds in situ in white lupin indicate that the GAs present in the phloem are derived mainly from the vegetative tissues of the plant. No evidence for metabolism of GAs in the phloem could be found.     

Vertical distribution of maize roots in relation to permanent soil characteristics

The vertical distribution of maize roots was studied in four contrasting soils, (arenosols, luvisols, planosols and vertisols) by using in-situ root mapping on vertical planes. The relationship between root contact frequency and depth was different for each soil, with a relatively low field-to-field variability within each soil type. The general aspect of this relationship did not change appreciably for three years in arenosols, with a low colonization in sandy layers probably being due to mechanical barriers. The relationship was consistently non-monotonic in luvisols and planosols, because of the sparse colonization of sandy layers. In planosols, these layers were traversed by some primary roots, which were essentially clustered in animal burrows. The distribution of root contact frequency was closer to an exponential function in vertisols. In these soils rooting depth and colonization of deep soil layers showed a marked increase during two dry years compared with a wet year. This was probably due to a denser net of shrinkage cracks and slickensides, where roots were essentially located in dry years. These results raise the possibility of modelling the decrease in root distribution with depth using soil information and climatic characteristics.     

Nuclear DNA amount,growth, and yield parameters in maize

Extensive nuclear DNA content variation has been observed inZea mays. Of particular interest is the effect of this variation on the agronomic potential of maize. In the present study, yield and growth data were collected on 12 southwestern US maize open-pollinated populations. These populations, originally cultivated by the Indians of the southwestern US for both human and animal consumption and adapted to various altitudes, were grown in replicated plots at the University of Illinois Agronomy-Plant Pathology South Farm. All growth and yield parameters were found to be negatively correlated with nuclear DNA amount. The negative correlations of nuclear DNA amount and growth parameters were more pronounced at 60 days after planting (DAP) than 30 DAP. Agronomically-important yield parameters, such as ear or seed weight and seed number per plant, also exhibited a significant negative correlations with nuclear DNA amount. These correlations demonstrate how the nucleotype may exhibit a high degree of influence on the agronomic phenotype. Although the results presented here represent only three replications at one location in 1 year, the observations noted suggest that nucleotype plays an integral role in determining the agronomic performance of maize. Further studies are needed to fully document this role.     

Increasing maize seed weight by enhancing the cytoplasmic ADP-glucose pyrophosphorylase activity in transgenic maize plants

ADP-glucose pyrophosphorylase (AGPase) plays a key role in regulating starch biosynthesis in cereal seeds and is likely the most important determinant of seed strength. The Escherichia coli mutant glgC gene (glgC16), which encodes a highly active and allosterically insensitive AGPase, was introduced into maize (Zea mays L.) under the control of an endosperm-specific promoter. Developing seeds from transgenic maize plants showed up to 2–4-fold higher levels of AGPase activity in the presence of 5 mM inorganic phosphate (Pi). Transgenic plants with higher cytoplasmic AGPase activity under Pi-inhibitory conditions showed increases (13–25%) in seed weight over the untransformed control. In addition, in all transgenic maize plants, the seeds were fully filled, and the seed number of transgenic plants had no significant difference compared with that of untransformed control. These results indicate that increasing cytoplasmic AGPase activity has a marked effect on sink activity and, in turn, seed weight in transgenic maize plants.     

Acclimation of white lupin to phosphorus deficiency involves enhanced expression of genes related to organic acid metabolism

White lupin (Lupinus albus L.) acclimates to phosphorus deficiency (–P) by the development of short, densely clustered lateral roots called proteoid (or cluster) roots. These specialized plant organs display increased exudation of citric and malic acid. The enhanced exudation of organic acids from P stressed white lupin roots is accompanied by increased in vitro phosphoenolpyruvate carboxylase (PEPC) and malate dehydrogenase (MDH) activity. Here we report the cloning of full-length white lupin PEPC and MDH cDNAs. RNA blot analysis indicates enhanced expression of these genes in –P proteoid roots, placing higher gene expression at the site of organic acid exudation. Correspondingly, macroarray analysis of about 1250 ESTs (expressed sequence tags) revealed induced expression of genes involved in organic acid metabolism in –P proteoid roots. In situ hybridization revealed that PEPC and MDH were both expressed in the cortex of emerging and mature proteoid rootlets. A C₃ PEPC protein was partially purified from proteoid roots of P deficient white lupin. Native and subunit Mr were determined to be 440 kD and 110 kD, respectively. Citrate and malate were effective inhibitors of in vitro PEPC activity at pH 7. Addition of ATP partially relieved inhibition of PEPC by malate but had little effect on citrate inhibition. Taken together, the results presented here suggest that acclimation of white lupin to low P involves modified expression of plant genes involved in carbon metabolism.     

Spatial and temporal variation in citrate and malate exudation and tissue concentration as affected by P stress in roots of white lupin

Exudation of carboxylates represents one the most efficient strategies used by P-starved white lupin (Lupinus albus L.) to acquire phosphorus from sparingly soluble sources. This exudation occurs through proteoid root clusters, with citrate being the predominant organic acid released. The occasional detection of malate in whole root exudates suggests that this acid would also be released, but from tissues other than root clusters. To investigate the spatial and temporal pattern of exudation, citrate and malate exudation and concentration were measured in whole roots and root sections of white lupin, from seedling emergence to plant senescence due to P starvation. Both organic acids were detected in whole root exudates of P-stressed plants, and they were released at similar rates throughout the experiment. Malate was predominantly exuded from apices of both seedling taproots and proteoid roots, whereas citrate exudation was restricted to proteoid root clusters. Studies directed to address the association between carboxylate exudation and concentration in proteoid root clusters showed a non-linear response for citrate, within the range of 7 to 23 mol g^–1 fresh weight. This association was further assessed by altering citrate concentration in the whole root. Adding P to 24-day-old P-starved plants reduced citrate concentration and exudation to the level of the control P-fed plants, demonstrating that citrate exudation and concentration are associated. Malate exudation and concentration did not correlate significantly. Results indicate that citrate release by P-starved white lupin would occur whenever a certain threshold of citrate concentration is attained, and that the sites, the rates and the span of transient exudation depend on the physiological age of the tissue.     

Treatment with salicylic acid decreases the effect of cadmium on photosynthesis in maize plants

The present study investigated the possible mediatory role of salicylic acid (SA) in protecting photosynthesis from cadmium (Cd) toxicity. Seeds of maize (Zea mays L., hybrid Norma) were sterilized and divided into two groups. Half of the seeds were presoaked in 500muM SA solution for only 6h, after which both groups were allowed to germinate for 3d and were then grown for 14d in Hoagland solution at 22/18 degrees C in a 16/8-h light/dark period and 120mumolm(-2)s(-1) PAR. All seedlings (without H(2)O and SA controls) were transferred to Cd-containing solutions (10, 15, and 25muM) and grown for 14d. The rate of CO(2) fixation and the activity of ribulose 1,5-bisphosphate carboxylase (RuBPC, EC 4.1.1.39) and phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) were measured. Changes in the levels of several important parameters associated with oxidative stress, namely H(2)O(2) and proline production, lipid peroxidation, electrolyte leakage, and the activities of antioxidative enzymes (superoxide dismutase (SOD, EC 1.15.1.1), ascorbate peroxidase (APX, EC 1.11.1.11), catalase (CAT, EC 1.11.1.6), and guaiacol peroxidase (POD, EC 1.11.1.7)) were measured. Exposure of the plants to Cd caused a gradual decrease in the shoot and root dry weight accumulation, with the effect being most pronounced at 25muM Cd. Seed pretreatment with SA alleviated the negative effect of Cd on plant growth parameters. The same tendency was observed for the chlorophyll level. The rate of CO(2) fixation was lower in Cd-treated plants, and the inhibition was partially overcome in SA-pretreated plants. A drop in the activities of RuBPC and PEPC was observed for Cd-treated plants. Pretreatment with SA alleviated the inhibitory effect of Cd on enzyme activity. Proline production and the rates of lipid peroxidation and electrolyte leakage increased in Cd-treated plants, whereas the values of these parameters were much lower in SA-pretreated plants. Treatment of plants with Cd decreased APX activity, but more than doubled SOD activity. Pretreatment with SA caused an increase in both APX and SOD activity, but caused a strong reduction in CAT activity. The data suggest that SA may protect cells against oxidative damage and photosynthesis against Cd toxicity.