Oxygen-Dependent Exclusion of Sodium Ions from Shoots by Roots of Zea mays (cv Pioneer 3906) in Relation to Salinity Damage

Using radio-tracers, we measured Na⁺ and K⁺ accumulation in roots and transport to shoots in Zea mays (cv Pioneer 3906) as a function of NaCl concentration and O₂ partial pressure in the nutrient solution. Under fully aerobic conditions, roots partially excluded Na⁺ from the shoots over a wide range of NaCl concentration (0.2-200 millimolar). With root anoxia, the exclusion mechanism broke down so that much greater amounts of Na⁺ reached the shoots, with simultaneous inhibition of K⁺ transport. The ratio Na⁺/K⁺ entering the shoot consequently increased 90 to 200 times. Increases in Na⁺ transport were first detected when the O₂ partial pressure was reduced from ambient (21% v/v) to 15%, whereas K⁺ transport was not inhibited until O₂ concentrations were <5%. Since soil O₂ deficiency can often accompany high salinity in irrigation agriculture, failure of the Na⁺ exclusion mechanism may be a contributory factor in salinity damage of salt-sensitive glycophytes.     

The Role of the Mesocotyl in Sodium Exclusion from the Shoot of Zea mays L. (cv. Pioneer 3906)

Drew, M. C. and Lauchli, A. 1986. The role of the mesocotylin sodium exclusion from the shoot of Zea mays L. (cv. Pioneer3906).—J. exp. Bot. 38: 409–418. The mesocotyl, located between the root and shoot, can stronglyaccumulate Na⁺ from the ascending transpiration stream, therebypotentially acting as a sink to protect the shoot from excessNa⁺. To determine the quantitative importance of the mesocotylas a Na⁺ sink, we grew plants with either short (9·0mm) or long(21 mm) mesocotyls, the latter resembling the sizefound in field-grown plants. At 13 d, plants were transferredfrom Na ⁺ -free nutrient solution to a ²²Na⁺ labelled solutionin which the concentration of NaCl was (mol m^–3) 1·0,10 or 100. The concentration of Na⁺ accumulated in the mesocotylin 24 h (g^–1 fr. wt.) exceeded that in the roots thatwere directly exposed to the nutrient solution. The amountsof ²²Na⁺ retained in the long mesocotyl were about double thatin the short ones and increased with time of exposure and NaClconcentration. At 1·0 and 10 mol m^–3 NaCl, theamounts of ²²Na⁺ retained in the mesocotyl were 6–19%of those reaching the shoot in 24 h, but with 100 mol m^–3NaCl, a damaging concentration for maize, this declined to 3–8%.The mesocotyl, even as a fully elongated structure is, therefore,unlikely to provide an appreciable alternative sink for Na⁺when NaCl reaches injurious concentrations. Key words: Ion transport, potassium, roots, salinity     

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 Deseeding on the Indole-3-acetic Acid Content of Shoots and Roots of Zea mays Seedlings

We wished to determine the effect of endosperm removal on the amounts of free and esterified indole-3-acetic acid (IAA) in young Zea mays seedlings. The increases of IAA derived from endosperm and from biosynthesis, but without correction for catabolic losses, were 0.9 picomole of free IAA per shoot per hour, and 1.1 picomoles per shoot per hour of ester IAA. After deseeding, free IAA in the shoot declines by 40% following kernel removal and total (free + ester) IAA declines at a rate of about 1 picomole per shoot per hour. A slight, but insignificant increase of ester IAA occurs following endosperm removal. In the primary roots, the decreases of free IAA and total (free + ester) IAA are accelerated by seed removal. Thus, the endosperm appears to be a major source of IAA for the shoot and root.     

The Release of Potassium and Sodium from Young Excised Roots of Zea mays under Various Efflux Conditions

The release of potassium and sodium from excised roots of Zea mays having similar contents of potassium and sodium was studied. At low temperature (2 C) the efflux rates of both cations were very similar, but at higher temperature (20 C) the potassium release was reduced considerably, whereas the sodium release was hardly affected. Also, under anaerobic conditions the potassium efflux rate was nearly as high as the sodium efflux rate, but with normal O₂ supply the potassium release was reduced to about one-fifth. Since a changing efflux medium compared with a constant efflux medium had no great influence upon the sodium release but influenced the potassium release very much, it is assumed that the low potassium release under normal metabolic conditions is due to a reabsorption of effluxed potassium from free space. For sodium this reabsorption is of minor significance, as the uptake potential of maize roots for sodium is very poor. It is concluded that the release of potassium and sodium is a diffusion process and that the cell membranes have rather similar diffusivities for these two cations.     

Electrotropism of Maize (Zea mays L.) Roots (Facts and Artifacts)

Intact and decapped primary roots of maize (Zea mays L.) were exposed to DC electric fields of 0.5 to 8.0 V/cm in low-salinity media to resolve conflicting results about the direction of electrotropism. In DC fields of 0.5 V/cm or 1.0 V/cm, intact roots always curved toward the cathode. In a field of 8.0 V/cm, intact roots curved toward the anode and stopped growth. Decapped roots also curved toward the anode both in weak and strong fields. The results indicate that growth toward the cathode is the true response of healthy roots.     

Defective Secretion of Mucilage is the Cellular Basis for Agravitropism in Primary Roots of Zea mays cv. Ageotropic

Root caps of primary, secondary, and seminal roots of Z. mayscv. Kys secrete large amounts of mucilage and are in close contactwith the root all along the root apex. These roots are stronglygraviresponsive. Secondary and seminal roots of Z. mays cv.Ageotropic are also strongly graviresponsive. Similarly, theircaps secrete mucilage and closely appress the root all alongthe root apex. However, primary roots of Z. mays cv. Ageotropicare non-responsive to gravity. Their caps secrete negligibleamounts of mucilage and contact the root only at the extremeapex of the root along the calyptrogen. These roots become graviresponsivewhen their tips are coated with mucilage or mucilage-like materials.Peripheral cells of root caps of roots of Z. mays cv. Kys containmany dictyosomes associated with vesicles that migrate to andfuse with the plasmalemma. Root-cap cells of secondary and seminal(i.e. graviresponsive) roots of Z. mays cv. Ageotropic are similarto those of primary roots of Z. mays cv. Kys. However, root-capcells of primary (i.e. non-graviresponsive) roots of Z. mayscv. Ageotropic have distended dictyosomal cisternae filled withan electron-dense, granular material. Large vesicles full ofthis material populate the cells and apparently do not fusewith the plasmalemma. Taken together, these results suggestthat non-graviresponsiveness of primary roots of Z. mays cv.Ageotropic results from the lack of apoplastic continuity betweenthe root and the periphery of the root cap. This is a resultof negligible secretion of mucilage by cells along the edgeof the root cap which, in turn, appears to be due to the malfunctioningof dictyosomes in these cells. Corn, dictyosomes, mucilage, root gravitropism, Zea mays cv. Ageotropic, Zea mays cv. Kys     

Ion Distribution in Salt-stressed Mature Zea mays Roots in Relation to Ultrastructure and Retention of Sodium

Ultrastructural features and the distribution of soluble ionshave been examined in mature roots of Zea mays plants grownin both NaCl and Na₂SO₄ salinities. When the plants were grown in either salt, the Na concentrationincreased proximally along the root with a concomitant declinein the K concentration. Both trends were reversed in the shoot. X-ray microanalysis of deep-frozen, fully hydrated specimensshowed that in salt-treated roots Na, and Cl, or S were distributedabout stoichiometrically in the cortex and endodermis. Na wasusually less concentrated than the anion in the lumens of thevessels, but was concentrated markedly relative to either Clor S in the adjoining xylem parenchyma cells. In the older, proximal parts of seminal roots of plants grownboth without salt (controls) and in the presence of either NaClor Na₂SO₄, wall developments occurred in xylem parenchyma cellsat the half-bordered pits in which the cell wall became markedlythicker and possessed a loosely packed fibrillar structure.These structures were not comparable with the transfer-celltype of protuberances reported in the roots of other species. In the xylem parenchyma of plants grown in the presence of Na₂SO₄there were dramatic increases in the quantities of rough endoplasmicreticulum, ribosomes, and mitochondria relative both to controlsand NaCl treatments. The results are discussed in relation to the possible functionof the xylem parenchyma of the mature root in the reabsorptionof Na from the xylem sap, which may mitigate adverse effectsof salinity in salt-sensitive glycophytes.     

Biochemical Characteristics of Membrane Fractions Isolated from Maize (Zea mays L.) Roots

Membrane fractions have been isolated from maize (Zea mays L.)roots by discontinuous density gradient centrifugation and phaseseparation methods. A number of approaches were tried with theaim of identifying specific membrane types, especially the plasmamembrane. These included the use of enzymic markers, determinationof glucose and leucine incorporation, separation of membraneproteins by SDS-PAGE, and attempts to identify the plasma membranefraction by cell surface labelling. The results are discussedin relation to the usefulness of membrane markers and the difficultiesof isolating surface membranes from higher plant tissues.     

Meiotic pairing in the hybrid (Zea diploperennis x Zea perennis) x Zea mays and its reciprocal

Genomic formulae, fertility, chromosome pairing, and the cryptic intergenomic pairing (induced by using diluted colchicine solution) were analysed in the tri-hybrid (MDP), obtained by crossing DP40 (2n=40, which was inferred in previous studies to have originated from the fusion of an unreduced gamete of Zea diploperennis with a normal gamete of Z. perennis) with the maize inbred line Zm40 (2n=40). MDP (2n=40) showed a higher fertility (90% of the seeds are viable) than Zm40 (60%) and DP40 (80%). A regular migration of 20 chromosomes to each pole occurred in 92% of the cells in anaphase I, while bridges were observed in the other 8% of the cells. When Zm40 was used as female of the crossing (Zm40 x DP40), ears were similar to corn. Conversely, ears resembled those of the wild species when cytoplasm was donoured by Zd. Then, it can be stated the existence of cytoplasmic influence on MDP ear type. MDP had almost no I or III, with an average of 0.04I + 10.90II + 0.01III + 4.50IV. The most frequent meiotic configuration was 10II + 5IV (43.93% of the cells). On average, 33.81 chiasmata/cell were observed (17.34, 0.05 and 16.42 average numbers of chiasmata/cell in bivalents, trivalents and tetravalents, respectively). It can be inferred that the 5IV were the product of homoeologous chromosome pairing of A genomes from the three species. On the other hand, the 10II configuration suggests separate pairing of the 5 homologous B chromosomes from maize and the 5 homoeologous B chromosomes from Zp and Zd.     

Collection of Gravitropic Effectors from Mucilage of Electrotropically-stimulated Roots of Zea mays L.

We placed agar blocks adjacent to tips of electrotropicallystimulated primary roots of Zea mays. Blocks placed adjacentto the anode-side of the roots for 3 h induced significant curvaturewhen subsequently placed asymmetrically on tips of vertically-orientedroots. Curvature was always toward the side of the root ontowhich the agar block was placed. Agar blocks not contactingroots and blocks placed adjacent to the cathode-side of electrotropicallystimulated roots did not induce significant curvature when placedasymmetrically on tips of vertically-oriented roots. Atomicabsorption spectrophotometry indicated that blocks adjacentto the anode-side of electrotropically-stimulated roots containedsignificantly more calcium than (1) blocks not contacting roots,and (2) blocks contacting the cathode-side of roots. These resultsdemonstrate the presence of a gradient of endogenous Ca in mucilageof electrotropically-stimulated roots (i.e. roots undergoinggravitropic-like curvature). Zea mays, corn, mucilage, root gravitropism, electrotropism     

Enhanced Mineral Uptake by Zea mays and Sorghum bicolor Roots Inoculated with Azospirillum brasilense

Inoculation of corn (Zea mays) seeds with Azospirillum brasilense strain Cd or Sp 7 significantly enhanced (30 to 50% over controls) the uptake of NO₃⁻, K⁺, and H₂PO₄⁻ into 3- to 4-day- and 2-week-old root segments. No gross changes in root morphology were observed; altered cell arrangement in the outer four or five layers of the cortex was seen in photomicrographs of cross sections of inoculated corn roots. The surface activity involved in ion uptake probably increased, as shown by the darker staining by methylene blue of the affected area. Shoot dry weight increased 20 to 30% in inoculated plants after 3 weeks, presumably by enhancement of mineral uptake. Corn and sorghum plants grown to maturity on limiting nutrients in the greenhouse showed improved growth from inoculation approaching that of plants grown on normal nutrient concentrations. Enhanced ion uptake may be a significant factor in the crop yield enhancement reported for Azospirillum inoculation.     

The Diversity of Archaea and Bacteria in Association with the Roots of Zea mays L.

The diversity of bacteria and archaea associating on the surface and interior of maize roots (Zea mays L.) was investigated. A bacterial 16S rDNA primer was designed to amplify bacterial sequences directly from maize roots by PCR to the exclusion of eukaryotic and chloroplast DNA. The mitochondrial sequence from maize was easily separated from the PCR-amplified bacterial sequences by size fractionation. The culturable component of the bacterial community was also assessed, reflecting a community composition different from that of the clone library. The phylogenetic overlap between organisms obtained by cultivation and those identified by direct PCR amplification of 16S rDNA was 48%. Only 4 bacterial divisions were found in the culture collection, which represented 27 phylotypes, whereas 6 divisions were identified in the clonal analysis, comprising 74 phylotypes, including a member of the OP10 candidate division originally described as a novel division level lineage in a Yellowstone hot spring. The predominant group in the culture collection was the actinobacteria and within the clone library, the a-proteobacteria predominated. The population of maize-associated proteobacteria resembled the proteobacterial population of a typical soil community within which resided a subset of specific plant-associated bacteria, such as Rhizobium- and Herbaspirillum-related phylotypes. The representation of phylotypes within other divisions (OP10 and Acidobacterium) suggests that maize roots support a distinct bacterial community. The diversity within the archaeal domain was low. Of the 50 clones screened, 6 unique sequence types were identified, and 5 of these were highly related to each other (sharing 98% sequence identity). The archaeal sequences clustered with good bootstrap support near Marine group I (crenarchaea) and with Marine group II (euryarchaea) uncultured archaea. The results suggest that maize supports a diverse root-associated microbial community composed of species that for the first time have been described as inhabitants of a plant-root environment.     

Characterization of Nitrate Reductases from Corn Leaves (Zea mays cv W64AxW182E) and Chlorella vulgaris: Sensitivity to a Proteinase Extracted from Corn Roots

The sensitivity of the two forms of nitrate reductase, NR_I and NR_II, obtained from the primary leaf of corn, to a limited action corn root proteinase has been examined. The corn inactivating protein (CIP) inhibited the overall reaction (NADH-NR) and the two partial reactions, cytochrome c reductase and reduced methyl viologen NR (MV-NR) of both forms of NR. NADH-cytochrome c reductase was more sensitive to the protease than MV-NR. NR_II was less sensitive to inactivation than NR_I. When NR_I and NR_II were inactivated and then subjected to native gel electrophoresis the protein bands associated with MV-NR activity shifted from an R_m value of 0.32 to 0.61 for NR_I and from an R_m of 0.28 to 0.60 for NR_II. For Chlorella NR these values are 0.32 and 0.70. The initial cleavage of the 116 kilodalton subunit of NR_I yielded fragments of 84 and 80 kilodaltons after a 5 minute incubation with CIP. With longer incubation times smaller fragments were also identified. For the Chlorella NR the initial cleavage products are approximately 68 and 25 kilodaltons. Longer incubation times also led to smaller fragments. The products of hydrolysis by this limited action protease are quite different for the corn and Chlorella NRs.     

Half-Life of sRNA from Primary Roots of Zea mays. A Contribution to the Cytokinin Production

Roots produce cytokinins, which could be generated from the catabolism of transfer RNA. To prove such a hypothesis, the half-life of sRNA from primary roots of corn was measured. The shortest half-life was estimated to be 3 days. Depending on growth, cells at the root tips are “moving” out of the growing region and become differentiated cells. Therefore the highest label at the root tip during the pulse incubation is moving backwards during the chase incubation. Prolonged chase increases the half-life of sRNA successively, possibly due to decreasing metabolic activities of the originally labeled cells.     

Gibberellin A(3) Is Biosynthesized from Gibberellin A(20) via Gibberellin A(5) in Shoots of Zea mays L

[17-¹³C,³H]-Labeled gibberellin A₂₀ (GA₂₀), GA₅, and GA₁ were fed to homozygous normal (+/+), heterozygous dominant dwarf (D8/+), and homozygous dominant dwarf (D8/D8) seedlings of Zea mays L. (maize). ¹³C-Labeled GA₂₉, GA₈, GA₅, GA₁, and 3-epi-GA₁, as well as unmetabolized [¹³C]GA₂₀, were identified by gas chromatography-selected ion monitoring (GC-SIM) from feeds of [17-¹³C, ³H]GA₂₀ to all three genotypes. ¹³C-Labeled GA₈ and 3-epi-G₁, as well as unmetabolized [¹³C]GA₁, were identified by GC-SIM from feeds of [17-¹³C, ³H]GA₁ to all three genotypes. From feeds of [17-¹³C, ³H]GA₅, ¹³C-labeled GA₃ and the GA₃-isolactone, as well as unmetabolized [¹³C]GA₅, were identified by GC-SIM from +/+ and D8/D8, and by full scan GC-MS from D8/+. No evidence was found for the metabolism of [17-¹³C, ³H]GA₅ to [¹³C]GA₁, either by full scan GC-mass spectrometry or by GC-SIM. The results demonstrate the presence in maize seedlings of three separate branches from GA₂₀, as follows: (a) GA₂₀ → GA₁ → GA₈; (b) GA₂₀ → GA₅ → GA₃; and (c) GA₂₀ → GA₂₉. The in vivo biogenesis of GA₃ from GA₅, as well as the origin of GA₅ from GA₂₀, are conclusively established for the first time in a higher plant (maize shoots).     

Effect of Endosperm Removal on 7 Normal NaOH-Labile Indole-3-acetic Acid Conjugates in Shoots and Roots of Zea mays Seedlings

The pool of amide-linked indole-3-acetic acid (amide IAA) in the shoot of growing etiolated seedlings of Zea mays increases between the 3rd and 5th day of germination to equal the amount of free IAA and two-thirds the amount of ester IAA. Deseeding the germinant changes the pool size of free and amide IAA in a manner suggestive of conversion of endogenous free IAA to amide IAA. Deseeding also caused an almost total disappearance of amide IAA from the root, demonstrating that the pool of amide IAA is not inert and can be actively metabolized in young Z. mays seedlings.     

Zinc Deficiency Affects the Levels of Endogenous Gibberellins in Zea mays L.

Zinc deficiency in Zea mays L. markedly reduced the level ofGA₁, but not GA₂₀, suggesting blockage of 3rß-hy-droxylation.The level of IAA was also decreased although not as markedly.Castasterone was affected less than IAA by zinc deficiency. (Received February 24, 1997; Accepted June 24, 1997)