Nitrate Reductase Activity in Maize (Zea mays L.) Leaves: I. Regulation by Nitrate Flux

The roles that leaf nitrate content and nitrate flux play in regulating the levels of nitrate reductase activity (NRA) were investigated in 8- to 14-day old maize (Zea mays L.) plants containing high nitrate levels while other environmental and endogenous factors were constant. The nitrate flux of intact plants was measured from the product of the transpiration rate and the concentration of nitrate in the xylem. NRA decreased when the seedlings were deprived of nitrate. The nitrate flux and the leaf nitrate content also decreased. When nitrate was resupplied to the roots, all three parameters increased.     

Regulation of Nitrate Reductase Activity in Corn (Zea mays L.) Seedlings by Endogenous Metabolites

Primary and secondary metabolites of inorganic nitrogen metabolism were evaluated as inhibitors of nitrate reductase (EC 1.6.6.1) induction in green leaf tissue of corn seedlings. Nitrite, nitropropionic acid, ammonium ions, and amino acids were not effective as inhibitors of nitrate reductase activity or synthesis. Increasing α-amino nitrogen and protein content of intact corn seedlings by culture techniques significantly enhanced rather than decreased the potential for induction of nitrate reductase activity in excised seedlings.     

Nitrate Reductase Activity in Maize (Zea mays L.) Leaves: II. Regulation by Nitrate Flux at Low Leaf Water Potential

Experiments were conducted to determine whether the nitrate flux to the leaves or the nitrate content of the leaves regulated the nitrate reductase activity (NRA) in leaves of intact maize (Zea mays L.) seedlings having low water potentials (ψ_w) when other environmental and endogenous factors were constant. In seedlings that were desiccated slowly, the nitrate flux, leaf nitrate content, and NRA decreased as ψ_w decreased. The decrease in nitrate flux was caused by a decrease in both the rate of transpiration and the rate of nitrate delivery to the transpiration stream. Upon rewatering, the recovery in NRA was correlated with the nitrate flux but not the leaf nitrate content.     

Variation in Nitrate Accumulation, Nitrate Reductase Activity and Nitrite Reductase Activity along Primary and Nodal Roots of Corn (Zea mays L.) Seedlings

Significant differences in NO₃ accumulation and nitrate reductaseactivity (NRA) were noted in the successive segments of developingyoung primary and nodal roots. This variation was also foundto be a function of root age. Nitrite reductase activity (NiRA)on the other hand had little variation among various segmentsof primary and nodal roots and also as a function of root age.These data suggest root NO₃ accumulation and root NRA are twoprocesses which are not directly linked. ¹ Present address: Division of Plant Physiology, Indian AgriculturalResearch Institute, New Delhi-110012, India. (Received December 3, 1983; Accepted June 18, 1983)     

Nitrate Reductase Activity and Polyribosomal Content of Corn (Zea mays L.) Having Low Leaf Water Potentials

Desiccation of 8- to 13-day-old seedlings, achieved by withholding nutrient solution from the vermiculite root medium, caused a reduction in nitrate reductase activity of the leaf tissue. Activity declined when leaf water potentials decreased below −2 bars and was 25% of the control at a leaf water potential of −13 bars. Experiments were conducted to determine whether the decrease in nitrate reductase activity was due to reduced levels of nitrate in the tissue, direct inactivation of the enzyme by low leaf water potentials, or to changes in rates of synthesis or decay of the enzyme.     

Acetolactate Synthase Activity in Developing Maize (Zea mays L.) Kernels

Acetolactate synthase (EC 4.1.3.18) activity was examined in maize (Zea mays L.) endosperm and embryos as a function of kernel development. When assayed using unpurified homogenates, embryo acetolactate synthase activity appeared less sensitive to inhibition by leucine + valine and by the imidazolinone herbicide imazapyr than endosperm acetolactate synthase activity. Evidence is presented to show that pyruvate decarboxylase contributes to apparent acetolactate synthase activity in crude embryo extracts and a modification of the acetolactate synthase assay is proposed to correct for the presence of pyruvate decarboxylase in unpurified plant homogenates. Endosperm acetolactate synthase activity increased rapidly during early kernel development, reaching a maximum of 3 micromoles acetoin per hour per endosperm at 25 days after pollination. In contrast, embryo activity was low in young kernels and steadily increased throughout development to a maximum activity of 0.24 micromole per hour per embryo by 45 days after pollination. The sensitivity of both endosperm and embryo acetolactate synthase activities to feedback inhibition by leucine + valine did not change during kernel development. The results are compared to those found for other enzymes of nitrogen metabolism and discussed with respect to the potential roles of the embryo and endosperm in providing amino acids for storage protein synthesis.     

Proliferation of Maize (Zea mays L.) Roots in Response to Localized Supply of Nitrate

Maize (Zea mays L.) plants with two primary nodal root axeswere grown for 8 d in flowing nutrient culture with each axisindependently supplied with . Dry matter accumulation by roots was similar whether 1.0 mol m^–3 was supplied to on( or both axes. When was supplied to only one axis, however, accumulationof dry matter within the root system was significantly greaterin the axis supplied with . The increased dry matter accumulation by the +N-treated axis was attributableentirely to increased density and growth of lateral branchesand not to a difference in growth of the primary axis. Proliferation of lateral branches for the + N axis was associatedwith the capacity for in situ reduction and utilization of aportion of the absorbed , especially in the apical region where lateral primordia are initiated. Althoughreduced nitrogen was translocated to the –N axis, concentrationsin the –N axis remained significantly lower than in the+N axis. The concentratio of reduced nitrogen, as well as invitro reductase activity, was greater in apical than in more basal regions of the +N axis. The enhancedproliferation of lateral branches in the + N axis was accompaniedby an increase in total respiration rate of the axis. Part ofthe increased respiration was attributable to increased massof roots. The specific respiration rate (umol CO₂ exolved perhour per gram root dry weight) was also greater for the +N thanfor the –N axis. If respiration rate is taken as representativeof sink demand, stimulation of initiation and growth of lateralsby in situ utilization of a localized exogenous supply of establishes an increased sink demand through enhancedmetabolic activity and the increased partitioning of assimilatesto the + N axis responds to the difference in sink demand between+N and –N axes. Key words: NO₃^- reduction, NO₃^- uptake nitrogen partitioning, root respiration, sink demand     

Endopolyploidy in Diploid and Tetraploid Maize (Zea mays L.)

Nuclei from different tissues such as stem, mesocotyl, nodalroot and root tip of diploid and tetraploid maize were isolated,stained with propidium iodide and passed through an EPICS-751flow-cytometer cell sorter. Variations in flow histograms wereobserved in different tissues. Stem tissues of both the diploidand tetraploid had two peaks representing G1 and G2 somaticnuclei. The remaining tissues in both the diploids and tetraploidsexhibited three peaks. The first peak observed in these tissuesrepresents G1 somatic nuclei of the lowest ploidy level. Thesecond peak represent G2 somatic nuclei of the lowest ploidylevel+G1 somatic nuclei of the next ploidy level. The thirdpeak represents G2 of the higher ploidy level+G1 somatic nucleiof the next higher ploidy level. Statistically significant differenceswere observed between the diploid and tetraploid maize tissueswith respect to nuclei distribution in the higher ploidy levelpeaks implying variation in the degree of endopolyploidy inthe diploid and tetraploid maize. The results of this studysuggest that the amount of endopolyploid observed in maize genotypeshas an effect on their overall agronomic performance under thefield conditions.Copyright 1993, 1999 Academic Press Zea mays L., maize, endopolyploidy, diploid, tetraploid, flow cytometry     

Fertility of Deep-frozen Maize (Zea mays L.) Pollen

On average, 50 per cent of maize pollen grains can be kept viableand almost 30 per cent remain fertile for up to a year whenthe water content on a fresh weight basis is reduced to about30 per cent of the original and the pollen is stored at –76or –196 °C. Over this period no significant differencewas found between storage at either temperature. Zea mays L., maize, pollen, fertility, viability     

Ethylene Evolution from Maize (Zea mays L.) Seedling Roots and Shoots in Response to Mechanical Impedance

The effect of mechanical impedance on ethylene evolution and growth of preemergent maize (Zea mays L.) seedlings was investigated by pressurizing the growth medium in triaxial cells in a controlled environment. Pressure increased the bulk density of the medium and thus the resistance to growth. The elongation of maize primary roots and preemergent shoots was severely hindered by applied pressures as low as 10 kilopascals. Following a steep decline in elongation at low pressures, both shoots and roots responded to additional pressure in a linear manner, but shoots were more severely affected than roots at higher pressures. Radial expansion was promoted in both organs by mechanical impedance. Primary roots typically became thinner during the experimental period when grown unimpeded. In contrast, pressures as low as 25 kilopascals caused a 25% increase in root tip diameter. Shoots showed a slight enhancement of radial expansion; however, in contrast to roots, the shoots increased in diameter even when growing unimpeded. Such morphological changes were not evident until at least 3 hours after initiation of treatment. All levels of applied pressure promoted ethylene evolution as early as 1 hour after application of pressure. After 1 hour, ethylene evolution rates had increased 10, 32, 70, and 255% at 25, 50, 75, and 100 kilopascals respectively, and continued to increase linearly for at least 10 hours. When intact corn seedlings were subjected to a series of hourly cycles of pressure, followed by relaxation, ethylene production rates increased or decreased rapidly, illustrating tight coupling between mechanical impedance and tissue response. Seedlings exposed to 1 microliter of ethylene per liter showed symptoms similar to those shown by plants grown under mechanical impedance. Root diameter increased 5 times as much as the shoot diameter. Pretreatment with 10 micromolar aminoethoxyvinyl glycine plus 1 micromolar silver thiosulfate maintained ethylene production rates of impeded seedlings at basal levels and restored shoot and root extension to 84 and 90% of unimpeded values, respectively. Our results support the hypothesis that ethylene plays a pivotal role in the regulation of plant tissue response to mechanical impedance.     

Effect of Exogenous Abscisic Acid on Proline Dehydrogenase Activity in Maize (Zea mays L.)

Plant responses to drought stress include proline and abscisic acid (ABA) accumulation. Proline dehydrogenase (PDH) (EC 1.4.3) is the first enzyme in the proline oxidation pathway, and its activity has been shown to decline in response to water stress (PJ Rayapati, CR Stewart [1991] Plant Physiol 95: 787-791). In this investigation, we determined whether ABA treatment affects PDH activity in a manner similar to drought stress in maize (Zea mays L.) seedlings. Four exogenous ABA treatments (0, 11, 33, and 100 micromolar ABA) were applied to well-watered maize seedlings. Mitochondria were isolated and PDH was solubilized using Nonidet P-40. PDH activity was measured by the reduction of iodonitrotetrazolium violet under proline-dependent conditions. There was no effect of ABA on PDH activity at 33 and 100 micromolar ABA, but there was a 38% decline at 11 micromolar. This decline was less than the 69% reduction in activity under drought stress. Endogenous ABA determinations and plant growth rate showed that ABA entered the plant and was affecting metabolic processes. ABA treatments had a small effect on shoot and root proline concentration, whereas drought stress caused a 220% increase in root tissues. We conclude that ABA is not part of the pathway linking drought stress and decreased PDH activity.     

Sugar Efflux from Maize (Zea mays L.) Pedicel Tissue

Sugar release from the pedicel tissue of maize (Zea mays L.) kernels was studied by removing the distal portion of the kernel and the lower endosperm, followed by replacement of the endosperm with an agar solute trap. Sugars were unloaded into the apoplast of the pedicel and accumulated in the agar trap while the ear remained attached to the maize plant. The kinetics of ¹⁴C-assimilate movement into treated versus intact kernels were comparable. The rate of unloading declined with time, but sugar efflux from the pedicel continued for at least 6 hours and in most experiments the unloading rates approximated those necessary to support normal kernel growth rates. The unloading process was challenged with a variety of buffers, inhibitors, and solutes in order to characterize sugar unloading from this tissue.

Unloading was not affected by apoplastic pH or a variety of metabolic inhibitors. Although p-chloromercuribenzene sulfonic acid (PCMBS), a nonpenetrating sulfhydryl group reagent, did not affect sugar unloading, it effectively inhibited extracellular acid invertase. When the pedicel cups were pretreated with PCMBS, at least 60% of sugars unloaded from the pedicel could be identified as sucrose. Unloading was inhibited up to 70% by 10 millimolar CaCl₂. Unloading was stimulated by 15 millimolar ethyleneglycol-bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetic acid which partially reversed the inhibitory effects of Ca²⁺. Based on these results, we suggest that passive efflux of sucrose occurs from the maize pedicel symplast followed by extracellular hydrolysis to hexoses.

     

Effects of Moniliformin on Mitosis in Maize (Zea mays L.)

Maize (Zea mays L. ‘Norfolk White’) roots were treatedwith solutions of moniliformin (a metabolite of Fusarium moniliformeSheldon) at 0.0001 M and 0.001 M for 8, 24, and 48 h. Only aslight inhibition of division was noted in root tips treatedwith the lower concentration. The higher concentration causeda disruption of the spindle apparatus and consequent C-mitosis,and metaphase accumulation. (Received February 14, 1984; Accepted May 18, 1984)     

Development of Endopeptidase Activities in Maize (Zea mays L.) Endosperms

An activity stain was used after native polyacrylamide gel electrophoresis, and at least 17 different endopeptidase activities were detected in maize (Zea mays L.) endosperm extracts prepared during the first 6 d after imbibition. The enzymes detected were classified into four groups based on their time of appearance and on their mobility in polyacrylamide gels. The first group, which included two enzymes present in dry endosperms, disappeared soon after imbibition. The second group, comprising five activity bands, appeared during the first 2 to 3 d after imbibition and then disappeared. The third set of enzymes increased continuously throughout the experimental period. The fourth group appeared after d 3 and remained at a constant level after that time. The endopeptidase activities were characterized by the effect of specific inhibitors on their activities. The two enzymes of the first group are metalloendopeptidases based on their sensitivity to ethylenediaminetetracetate (EDTA). Enzymes of the second, third, and fourth groups are sulfhydryl-endopeptidases as judged by their sensitivity to antipain, chymostatin, leupeptin, and E-64 and by their requirement for 2-mercaptoethanol. Pepstatin, phenylmethylsulfonyl fluoride, or EDTA had no effect on these enzymes. Many of the second, third, and fourth group enzymes cleaved [alpha]-zein-rich proteins as well as such easily obtained proteins as gelatin (used in our standard assay) and hemoglobin. The second group had a high affinity for [gamma]-zein, whereas none of the bands in the fourth group of enzymes cleaved this type of zein. The two metalloenzymes of the first group cleaved neither [alpha]- nor [gamma]-zeins.     

Anatomy of Seedling Roots of Tropical Maize (Zea mays L.) Cultivars at Low Water Supply

Establishment of maize seedlings can be difficult at low soilmoisture content. Anatomy of root metaxylem vessels may influencethe capacity for water transport and respective genotypic differencesmight be useful for selection purposes. To test this, six tropicalmaize (Zea mays L.) cultivars were grown in large PVC tubescontaining a sandy substrate at 5% (M5) and 10% (M10) moisturecontents for 2 weeks. The percentage changes in root diametersdue to M5 was similar for most cultivars but differed for mainroot types. Root diameters were not consistently related tometaxylem structure, but in a few cases, thin roots had smallerdiameter metaxylem vessels. The M5 treatment reduced the numberof late metaxylem vessels of primary roots by about 0 to 20%,while effects on nodal roots were slight. Generally, the ratioof cross-sectional areas between late and early metaxylem vesselsincreased from primary to seminal and nodal roots. Within thecultivar Tuxpefio this ratio was much reduced by M5. A few cultivarsmaintained the combined cross-sectional areas of metaxylem vesselsat M5 in some main root types, but only one cultivar could achievethis for the total of cross-sectional areas of metaxylem vessels,calculated over all root axes, by increasing the number of seminaland nodal roots. These anatomical traits seemed to be mostlyconstitutive with limited response to an actual environment,but they could be decisive for the suitability of a cultivarto an environment with frequent water shortages during seedlingestablishment. Key words: Metaxylem vessels, water stress, tropical maize     

Purification and Partial Characterization of Maize (Zea mays L.) beta-Glucosidase

Maize (Zea mays L.) β-glucosidase (β-d-glucoside glucohydrolase, EC 3.2.1.21) was extracted from the coleoptiles of 5- to 6-day-old maize seedlings with 50 millimolar sodium acetate, pH 5.0. The pH of the extract was adjusted to 4.6, and most of the contaminating proteins were cryoprecipitated at 0°C for 24 hours. The pH 4.6 supernatant from cryoprecipitation was further fractionated by chromatography on an Accell CM column using a 4.8 to 6.8 pH gradient of 50 millimolar sodium acetate, which yielded the enzyme in two homogeneous, chromatographically different fractions. Purified enzyme was characterized with respect to subunit molecular weight, isoelectric point, amino acid composition, NH₂-terminal amino acid sequence, pH and temperature optima, thermostability, and activity and stability in the presence of selected reducing agents, metal ions, and alkylating agents. The purified enzyme has an estimated subunit molecular mass of 60 kilodaltons, isoelectric point at pH 5.2, and pH and temperature optima at 5.8 and 50°C, respectively. The amino acid composition data indicate that the enzyme is rich in Glx and Asx, the sum of which approaches 25%. The sequence of the first 20 amino acids in the N-terminal region was H₂N-Ser-Ala-Arg-Val-Gly-Ser-Gln-Asn-Gly-Val-Gln-Met-Leu-Ser-Pro-(Ser?) -Glu-Ile-Pro-Gln, and it shows no significant similarity to other proteins with known sequence. The enzyme is extremely stable at 0 to 4°C up to 1 year but loses activity completely at and above 55°C in 10 minutes. Likewise, the enzyme is stable in the presence of or after treatment with 500 millimolar 2-mercaptoethanol, and it is totally inactivated at 2000 millimolar 2-mercaptoethanol. Such metal ions as Hg²⁺ and Ag⁺ reversibly inhibit the enzyme at micromolar concentrations, and inhibition could be completely overcome by adding 2-mercaptoethanol at molar excess of the inhibitory metal ion. The alkylating agents iodoacetic acid and iodoacetamide irreversibly inactivate the enzyme and such inactivation is accelerated in the presence of urea.     

不同基因型玉米间作的群体质量

采用大田试验,研究了不同基因型玉米间作的群体质量特征.结果表明,HF9‖XD20间作,植株中部叶片平均叶龄延长,而对下部和上部叶片影响不大;ZD958‖LD981间作,ZD958植株下部叶片平均叶龄延长,而中、上部叶片则缩短,LD981植株下、中、上部叶片平均叶龄均有所延长.吐丝前,群体叶面积指数(LAI)单间作无明显差异,吐丝后,HF9和LD981的LAI分别大于和显著大于单作群体,而ZD958和XD20则分别小于和显著小于单作群体.紧凑型品种和半紧凑型品种间作增加了群体透光率,吐丝后10d,4个品种棒三叶叶色值(SPADR)均有所增加,并且除ZD958外,其余3个品种棒三叶净光合速率均有所增加,其中LD981增加显著.间作对吐丝以前的群体干物质积累量影响不大,吐丝后,半紧凑型品种(HF9和LD981)的干物质积累量增加,其中LD981增加显著,而紧凑型品种(XD20和ZD958)的干物质积累量减少,其中ZD958显著减少;间作还提高了收获指数,并且两种间作群体的土地当量比(LER)均大于1.结果提示,紧凑型与半紧凑型玉米品种的间作可以提高群体质量,延长叶片功能期,提高光合效率,增加籽粒产量.     

Further Studies on Mosaic Disease of Maize (Zea mays L.)

Schizaphis graminum (Rondani) is proved to be an additional vector of maize mosaic virus (MMV). The pH range for the infectivity of the virus in extracted juice is found to be from 4.4 to 9.0, the optimum being 5.6 to 7.2. Effect of certain chemicals on the virusin vitro has also been studied. Cross protection between MMV and Sugar-cane mosaic virus (SMV) indicated positive results. It has been concluded on the basis of similar physical properties, tolerance towards certain chemicals, host range, symptomatology, aphid vectors and positive immunological tests, that MMV and SMV are related viruses.     

Bacterial Streak Disease of Maize (Zea mays L.) in South Africa

Bacterial streak disease of maize is currently causing some concern among breeders in South Africa. The causal organism of this previously undescribed disease was successfully isolated and its pathogenicity established using KoCH's postulates. Standard physiological and biochemical tests used to identify phytopathogenic bacteria indicated that the bacterium is a Xanthomonas campestris pathovar. Comparisons between this organism and other recognized X. campestris pathovars of the Poaceae indicated that apart from some minor differences the maize streak pathogen is physiologically similar to X. campestris pv. holcicola. However, in repeated reciprocal inoculation experiments all attempts to induce disease symptoms in sorghum with the maize streak pathogen were unsuccessful. Conversely, X. campestris pv. holcicola did produce symptoms in maize leaves. In all the maize cultivars tested the symptoms produced by the maize streak pathogen were, however, always considerably more severe than those caused by X. campestris pv. holcicola. Notwithstanding its physiological similarity to X. campestris pv. holicola it would appear that on the grounds of host specificity the maize streak pathogen warrants new pathovar status. The name X. campestris pv. zeae is proposed.