Hypoxic Induction of Anoxia Tolerance in Root Tips of Zea mays

When root tips of fully aerobic, intact maize (Zea mays L.) seedlings are made anaerobic, viability normally is only 24 hours or less at 25°C. We find that viability can be extended to at least 96 hours if seedlings are given a hypoxic pretreatment for 18 hours by sparging the solution with 4% O₂ in nitrogen (v/v) before anoxia. Fully aerobic root tips (sparged with 40% O₂) had very low alcohol dehydrogenase (ADH) activity (per gram root fresh weight), and the level remained low under anoxia. In hypoxically pretreated roots, however, high levels of ADH activity were induced, and activity rose further during the initial 24 hours of anoxia, and then remained high at about 20 times that of controls in 40% O₂. ADH activity in roots in solution sparged with air (21% O₂) was about three times that in 40% O₂. Improved viability of hypoxically pretreated root tips was associated with maintenance of a high energy metabolism (ATP concentration, total adenylates, and adenylate energy charge). Roots that were not pretreated lost 94% of the total adenylates and ATP at 24 hours of anoxia. The relation between induced ADH activity, energy metabolism, and improved anoxia-tolerance in acclimated maize root tips is discussed.     

Root Growth Inhibitors from Root Cap and Root Meristem of Zea mays L.

A micro-assay based on the growth inhibition of root segmentsof the seminal roots of Zea mays has been used to investigatethe root-growth-inhibiting substances in root caps and meristemsrespectively of the roots of Zea mays. This micro-assay is sensitiveto 50 pg of IAA or less. Paper chromatography of the acid fractionof methanolic extracts shows the presence of one main inhibitorin root caps and a different main inhibitor in root meristems.Neither is IAA, whose presence in meristems is sometimes indicatedby small inhibitions (or stimulations) at the characteristicRf of IAA. A Commelina leaf-epidermis assay shows the presenceof one stomata-closing ABA-like substance in root caps and onein meristems, one corresponding in Rf to the main root-growthinhibitor from the root cap. The implications of these findingsfor the geotropic responses of roots is briefly discussed.     

Root Growth, Secondary Root Formation and Root Gravitropism in Carotenoid-deficient Seedlings of Zea mays L.

The effect of ABA on root growth, secondary-root formation androot gravitropism in seedlings of Zea mays was investigatedby using Fluridone-treated seedlings and a viviparous mutant,both of which lack carotenoids and ABA. Primary roots of seedlingsgrown in the presence of Fluridone grew significantly slowerthan those of control (i.e. untreated) roots. Elongation ofFluridone-treated roots was inhibited significantly by the exogenousapplication of 1 mM ABA. Exogenous application of 1 µMand 1 nM ABA had either no effect or only a slight stimulatoryeffect on root elongation, depending on the method of application.The absence of ABA in Fluridone-treated plants was not an importantfactor in secondary-root formation in seedlings less than 9–10d old. However, ABA may suppress secondary-root formation inolder seedlings, since 11-d-old control seedlings had significantlyfewer secondary roots than Fluridone-treated seedlings. Rootsof Fluridone-treated and control seedlings were graviresponsive.Similar data were obtained for vp-9 mutants of Z. mays, whichare phenotypically identical to Fluridone-treated seedlings.These results indicate that ABA is necessary for neither secondary-rootformation nor for positive gravitropism by primary roots. Zea mays, gravitropism, carotenoid-deficient, Fluridone, root growth, vp-9 mutant     

The Effects of Mechanical Impedance to Growth on the Levels of ABA and IAA in Root Tips of Zea mays L.

Extraction and analytical methods have been refined and newones devised to allow precise determinations by GC-EC of thelevels of abscisic acid (ABA) and indol-3ylacetic acid (IAA)in samples of maize root tips as small as 1.0 g fr. wt. Seminalroots of 5-d-old maize seedlings grown in normal (bulk density1200 kg m^–3) and compacted (bulk density 1600 kg m^–3)sand/garden loam mixtures have been examined. Seminal rootsfrom compacted soil had an average length of about 40% of thatof control roots and were much thicker. The ABA levels in 10mm tips of impeded roots (c. 25–35 ng g^–1 fr.wt.)did not differ significantly from those of normal root tipson both a fresh and dry weight basis. The levels in 0–1mm tips were approximately double those in the remaining 1–10mm zones. IAA levels were increased by about 3 times in impededroots (176.3 as compared with 52.4 ng g^–1 fr.wt) and itis concluded that this response is likely to be the main causeof the morphological and growth changes brought about by soilcompaction.     

The Exchange of sodium Ions in the Root of Zea mays

The apparent cation-exchange capacity of maize roots has beenestimated in 100 mM NaCL and I mM NaCl by observing the effluxkinetics of isotopically labelled sodium from this tissue. Thisapproach indicated that the apparent capacity was 70 m.equiv/kgof tissue (fresh weight) and 2 m.equiv/kg of tissue (fresh weight)in 100 mM and I mM NaCl respectively; the pH of the medium was5.8 in each case.Similar sodium efflux experiments were performedon isolated stelar tissue from these roots and such experimentsgave an apparent exchange capacity of 86 m.equiv/kg of tissue(fresh weight) and 6 m.equiv/kg of tissue (fresh weight) in100 mM and I mM NaCl respectively, both at pH 5.8.The dependenceof the magnitude of the apparent exchange capacity upon theelectrolyte concentration of the external medium is discussed.     

Effects of Salinity on Water Transport of Excised Maize (Zea mays L.) Roots

The root pressure probe was used to determine the effects of salinity on the hydraulic properties of primary roots of maize (Zea mays L. cv Halamish). Maize seedlings were grown in nutrient solutions modified by additions of NaCl and/or extra CaCl₂ so that the seedlings received one of four treatments: Control, plus 100 millimolar NaCl, plus 10 millimolar CaCl₂, plus 100 millimolar NaCl plus 10 millimolar CaCl₂. The hydraulic conductivities (Lp_r) of primary root segments were determined by applying gradients of hydrostatic and osmotic pressure across the root cylinder. Exosmotic hydrostatic Lp_r for the different treatments were 2.8, 1.7, 2.8, and 3.4·10⁻⁷ meters per second per megapascals and the endosmotic hydrostatic Lp_r were 2.4, 1.5, 2.7, and 2.3·10⁻⁷ meters per second per megapascals, respectively. Exosmotic Lp_r of the osmotic experiments were 0.55, 0.38, 0.68, and 0.60·10⁻⁷ meters per second per megapascals and the endosmotic Lp_r were 0.53, 0.21, 0.56, and 0.54·10⁻⁷ meters per second per megapascals, respectively. The osmotic Lp_r was significantly smaller (4-5 times) than hydrostatic Lp_r. However, both hydrostatic and osmotic Lp_r experiments showed that salinization of the growth media at regular (0.5 millimolar) calcium levels decreased the Lp_r significantly (30-60%). Addition of extra calcium (10 millimolar) to the salinized media caused ameliorative effects on Lp_r. The low Lp_r values may partially explain the reduction in root growth rates caused by salinity. High calcium levels in the salinized media increased the relative availability of water needed for growth. The mean reflection coefficients of the roots using NaCl were between 0.64 and 0.73 and were not significantly different for the different treatments. The mean values of the root permeability coefficients to NaCl of the different treatments were between 2.2 and 3.5·10⁻⁹ meters per second and were significantly different only in one of four treatments. Cutting the roots successively from the tip and measuring the changes in the hydraulic resistance of the root as well as staining of root cross-sections obtained at various distances from the root tip revealed that salinized roots had mature xylem elements closer to the tip (5-10 millimeters) compared with the controls (30 millimeters). Our results demonstrate that salinity has adverse effects on water transport and that extra calcium can, in part, compensate for these effects.     

Effect of Low Root Medium pH on Net Proton Release, Root Respiration, and Root Growth of Corn (Zea mays L.) and Broad Bean (Vicia faba L.)

The effect of low pH on net H⁺ release and root growth of corn (Zea mays L.) and broad bean (Vicia faba L.) seedlings was investigated in short-term experiments at constant pH. Broad bean was more sensitive to low pH than corn: the critical values (pH values below which net H⁺ release and root growth ceased) were pH 4.00 (broad bean) and pH 3.50 (corn) at 1 millimolar Ca²⁺. Both proton release and root growth were progressively inhibited as the medium pH declined. Additional Ca²⁺ in the root medium helped to overcome the limitations of low pH for net H⁺ release and root growth. Potassium (for corn) and abscisic acid (for broad bean) increased both net H⁺ release and root growth rate at the critical pH value. It is concluded that poor root growth at low pH is caused by a lack of net H⁺ release that may decrease cytoplasmic pH values. Inhibited net H⁺ release at high external H⁺ activity is not due to a shortage of energy supply to the H⁺ ATPase. Instead, a displacement of Ca²⁺ by H⁺ at the external side of the plasmalemma may enhance reentry of H⁺ into root cells.     

The Effects of Rootzone Salinity and Hypoxia on Shoot and Root Growth in Trifolium Species

The effect of concurrent salinity (0-60 mM NaCl) and rootzonehypoxia (flooding for up to 15 d) on shoot and root growth andshoot ion concentrations of six species of Trifolium (T. subterraneumL., T. fragiferum L., T. michelianum Savi., T. isthmocarpumBot., T. purpureum Lois., and T. repens L.), was studied intwo greenhouse experiments. There was a significant salinityx flooding effect for shoot yield but no significant salinityx flooding x species interaction although individual speciesdiffered significantly (P < 0·001) in their growthresponse to the saline or flooded conditions separately. Concentrationsof Na and Cl in the shoots of all species increased with increasingperiods of saline flooding and there was a significant salinityx flooding interaction. Sodium and Cl concentrations were significantlyhigher (P < 0·001) in T. purpureum, the species inwhich shoot growth was most depressed by saline flooding, thanother species. In T. michelianum, T. fragiferum and T. repens,fresh and dry weight of roots increased with flooding underboth saline and non-saline conditions while in T. subterraneumroot growth decreased. A significant proportion of the increasedroot growth in the first three species occurred as new adventitiousroots. These roots had higher percentages of internal gas spaceswithin the root tissue even in the presence of salinity comparedwith roots from non-flooded conditions. There were also significantlymore gas spaces in the total root tissue in T. fragiferum andT. repens under saline-flooding than in roots of T. subterraneum.Electron micrographs of the root cross sections illustratedthe presence of these gas spaces or aerenchyma. Trifolium fragiferum, T. repens and T. michelianum are morelikely to be suited to growth in soils prone to high salinityand to flooding than are T. subterraneum, T. purpureum and T.ishmocarpum.Copyright 1993, 1999 Academic Press Trifolium subterraneum, Trifolium fragiferum, Trifolium michelianum, Trifolium isthmocarpum, Trifolium purpureum, Trifolium repens, salinity, flooding, hypoxia, adventitious roots, aerenchyma, subterranean clover, white clover, strawberry clover, purple clover, balansa clover     

The Effect of Light Intensity and Relative Humidity on Growth Rate and Root Respiration of Plantago lanceolata and Zea mays

Plants of Plantago lanceolata L. and Zea mays L., cv. ‘Campo’were grown at two levels of light intensity. Especially in theroots, the rate of dry matter accumulation decreased at lowlight intensity. The carbohydrate content of both roots andshoots of P. lanceolata was not affected by light intensity.The relative contribution of SHAM¹-sensitive respiration, thealternative chain, to total root respiration of both P. lanceolataand Z. mays, was not affected by light intensity during thedaytime. The alternative pathway was somewhat decreased at theend of the dark period, but not in the root tips (0–5mm) where it still contributed 56% in respiration. It was, therefore,concluded that photosynthesis is not a major factor in regulationof root growth in the species investigated. To see whether the effect of light intensity on root growthrate was via transpiration, plants of Z. mays were grown atdifferent air humidities. Both high humidity and low light intensityaffected the root morphology in such a way that the distancebetween the apex and the first laterals on the primary rootaxis increased. It is suggested that this effect on root morphologyis due to transpiration and the subsequent removal of root-producedinhibitors of lateral root growth; although light intensityalso affected the rate of dry matter accumulation of roots andthe rate was not affected by the humidity of the air. It is,therefore, concluded that the effect of light intensity on therate of dry matter accumulation of roots of Z. mays is not viaan effect on transpiration.     

Effects of Cytokinins on the Growth of Primary Roots of Zea mays

Intact primary roots of Zea mays seedlings, apical 6-mm segmentsisolated from the intact primary roots and 5-mm detipped segments,prepared from the 6-mm apical segments by removal of the apical1-mm meristematic region, were incubated in potassium-phosphatebuffer that contained various concentrations of kinetin, 6-benzylaminopurine(BAP) or zeatin. These cytokinins inhibited the growth of intactprimary roots but they promoted the growth of both tipped anddetipped apical segments. In other words, they promoted thegrowth of root segments irrespective of the presence or absenceof apical meristematic regions. Detipped segments were stoodvertically, with their apical or basal cutends in contact withan agar plate that contained the abovementioned buffer and variousconcentrations of kinetin, BAP or zeatin so that cytokininswere supplied either from the apical or basal cut-ends. Cytokininssupplied from the top promoted the growth of the segments, whilethose supplied from the base did not. These results indicate that the response of roots to the exogenouslyapplied cytokinins is not influenced by the presence of theroot meristem but is significantly affected by the way in whichcytokinins are supplied. (Received October 17, 1995; Accepted August 28, 1995)     

K+-Na+ Exchange and Selectivity in Barley Root Cells: Effect of Na+ on the Na+ Fluxes

Using the compartmental analysis the unidirectional Na⁺ fluxesin cortical cells of barley roots, the cytoplasmic and vacuolarNa⁺ contents Q_c and Q_v, and the trans-root Na⁺ transport R'have been studied as a function of the external Na⁺ concentration.Using the re-elution technique the effect of low K⁺ concentrationson the plasmalemma efflux _co of Na⁺ (K+-Na+ exchange) and onR' was investigated at different Na+ concentrations and correspondinglydifferent values of the cytoplasmic sodium content Qc. The relationof the K+-dependent Na+ efflux coK+-dep to Qc or to the cytoplasmicNa+ concentration obeyed Michaelis-Menten kinetics. This isconsistent with a linkage of co, K+-dep to K+ influx by a K⁺-Na⁺exchange system. The apparent Km corresponded to a cytoplasmicNa⁺ concentration of 28 mM at 0·2 mM K⁺ and about 0·2mM Na⁺ in the external solution. 0·2 mM K⁺ stimulatedthe plasma-lemma efflux of Na⁺ and inhibited Na⁺ transport selectivelyeven in the presence of 10 mM Na⁺ in the external medium showingthe high efficiency of the K⁺-Na⁺ exchange system. However,_co, K⁺-dep was inhibited at 10 mM Na¹ compared to lower Na¹concentrations suggesting some competition of Na¹ with K¹ atthe external site of the exchange system. The effect of theNa+ concentration on Na¹ influx _oc is discussed with respectto kinetic models of uuptake.     

The Effect of High Concentrations of Heavy Water on Root Morphogenesis in Zea mays

The effect of high concentrations of heavy water on young rootsof Zea mays was investigated. Submersion for 24 hrs. in solutionsof 80–90 per cent. D₂O will temporarily stop growth ofthe primary root. During the treatment period the root-tip regionswill swell as the result of radial cell enlargement, primarilyin the cortical region. Upon removal from D₂O growth will resume,initially at a slower rate than controls but ultimately at similarmaximum rates. Lateral root production is inhibited in thatportion of the primary root which is the region of elongationat time of treatment. However, in the swollen region lateralroot formation is accentuated. The mitotic index drops sharplyupon introduction of the primary roots into 80 per cent. D₂O.Within 12 hrs. the index drops to zero and remains at this leveluntil the roots are removed from the heavy water. The mitoticindex then rises again, reaching control values within 24 hrs.It is suggested that the gentle, temporary inhibitory actionof D₂O makes this substance a useful tool in morphogenetic investigations.     

Studies on the Growth and Indole-3-Acetic Acid and Abscisic Acid Content of Zea mays Seedlings Grown in Microgravity

Measurements were made of the fresh weight, dry weight, dry weight-fresh weight ratio, free and conjugated indole-3-acetic acid, and free and conjugated abscisic acid in seedlings of Zea mays grown in darkness in microgravity and on earth. Imbibition of the dry kernels was for 17 h prior to launch. Growth was for 5 d at ambient orbiter temperature and at a chronic accelerational force of the order of 3 × 10⁻⁵ times earth gravity. Weights and hormone content of the microgravity seedlings were, with minor exceptions, not statistically different from seedlings grown in normal gravity. The tissues of the shuttle-grown plants appeared normal and the seedlings differed only in the lack of orientation of roots and shoots. These findings, based upon 5 d of growth in microgravity, cannot be extrapolated to growth in microgravity for weeks, months, and years, as might occur on a space station. Nonetheless, it is encouraging, for prospects of bioregeneration of the atmosphere and food production in a space station, that no pronounced differences in the parameters measured were apparent during the 5 d of plant seedling growth in microgravity.     

Free Radicals in the Root System and Stem of Zea mays

The concentration of free radicals in different parts of theroot system and the stem of 130, 158 and 165 d old maize plantswere measured. The highest concentrations of free radicals werefound in the tissue of the primary root and in the adventitiousroots of the 1st and 2nd nodes. In the stem tissues, the highestconcentration of free radicals was found in the tissues of the14th to 15th internode. The concentration of free radicals inthe tissues of the nodal roots decreased with increasing nodenumber, whereas in the tissues of the overground stem it increasedwith internode number. This pattern may be correlated with themetabolic activity of tissues.     

Chimeras and the Origin of Lateral Root Primordia in Zea mays

The difficulty of determining the contribution made by the pericycleand endodermis of mother roots to lateral primordia in the Gramineaehas been solved by inducing polyploid chimeras at initiation. The endodermis forms a layer covering the primordium, but thislayer does not form the epidermis of the lateral. It does formthe root cap of the young primordium, but this is replaced ata variable stage of development by the quiescent centre donatinga new set of cap initials of pericyclic origin. Reasons forthe previous diversity of interpretations are presented. chimera, lateral root primordia, Zea mays     

Effects of a Magnetic Field on the Growth of Primary Roots of Zea mays

Caryopses with primary roots of Zea mays L. (cv. Golden CrossBantam 70) were incubated on agar-solidified distilled water(0.4% agar) in a magnetic field of 5 k gauss or 0.01 k gauss(control), the direction of root growth corresponding to thedirection of magnetic field from the north- to the south-seekingpole. The rate of growth of the roots exposed to 5 k gauss wasincreased by about 25% over that of the controls (0.01 k gauss).When caryopses with primary roots were incubated on agar-solidifieddistilled water that had previously been exposed to a magneticfield of 5 k gauss or 0.01 k gauss, no differences in ratesof root growth were observed. The growth rate of the primaryroot increased with increased magnetic flux density (from 0.01k to 5 k gauss). The orientation of the root in terms of thedirection of the magnetic field (from the north- to the south-seekingpole) affected the rate of growth of the root. When the directionof root growth was in line with the direction of the magneticfield of 5 k gauss or in the direction opposite to that of thefield, growth rates increased by 27% and 22%, respectively,of the growth rate of the controls (magnetic field of 0.01 kgauss). When the direction of growth was perpendicular to thedirection the field, the growth rate increased by 15% of thatof the control (0.01 k gauss). It appears that a magnetic stimulusmay induce an increase in the rate of root growth in some plantmaterials. (Received March 23, 1988; Accepted August 9, 1988)     

Responses by Coleoptiles of Intact Rice Seedlings to Anoxia: K+ Net Uptake from the External Solution and Translocation from the Caryopses

This study evaluated the effects of anoxia on K⁺ uptake andtranslocation in 3–4-d-old, intact, rice seedlings (Oryzasativa L. cv. Calrose). Rates of net K⁺ uptake from the mediumover 24 h by coleoptiles of anoxic seedlings were inhibitedby 83–91 %, when compared with rates in aerated seedlings.Similar uptake rates, and degree of inhibition due to anoxia,were found for Rb⁺ when supplied over 1·5–2 h,starting 22 h after imposing anoxia. The Rb⁺ uptake indicatedthat intact coleoptiles take up ions directly from the externalsolution. Monovalent cation (K⁺ and Rb⁺) net uptake from thesolution was inhibited by anoxia to the same degree for thecoleoptiles of intact seedlings and for coleoptiles excised,‘aged’, and supplied with exogenous glucose. Transportof endogenous K⁺ from caryopses to coleoptiles was inhibitedless by anoxia than net K⁺ uptake from the solution, the inhibitionbeing 55 % rather than 87 %. Despite these inhibitions,osmotic pressures of sap (_sap) expressed from coleoptiles ofseedlings exposed to 48 h of anoxia, with or without exogenousK⁺, were 0·66 ± 0·03 MPa; however,the contributions of K⁺ to _sap were 23 and 16 %, respectively.After 24 h of anoxia, the K⁺ concentrations in the basal10 mm of the coleoptiles of seedlings with or without exogenousK⁺, were similar to those in aerated seedlings with exogenousK⁺. In contrast, K⁺ concentrations had decreased in aeratedseedlings without exogenous K⁺, presumably due to ‘dilution’by growth; fresh weight gains of the coleoptile being 3·6-to 4·7-fold greater in aerated than in anoxic seedlings.Deposition rates of K⁺ along the axes of the coleoptiles werecalculated for the anoxic seedlings only, for which we assessedthe elongation zone to be only the basal 4 mm. K⁺ depositionin the basal 6 mm was similar for seedlings with or withoutexogenous K⁺, at 0·6–0·87 µmolg^–1 f. wt h^–1. Deposition rates in zones above6 mm from the base were greater for seedlings with, thanwithout, exogenous K⁺; the latter were sometimes negative. Weconclude that for the coleoptiles of rice seedlings, anoxiainhibits net K⁺ uptake from the external solution to a muchlarger extent than K⁺ translocation from the caryopses. Furthermore,K⁺ concentrations in the elongation zone of the coleoptilesof anoxic seedlings were maintained to a remarkable degree,contributing to maintenance of _sap in cells of these elongatingtissues.