Root growth responses to lead in young maize seedlings

This work was undertaken to follow the appearance and development of symptoms of lead toxicity in growing roots of seedlings. The effects of lead nitrate (10^-2–10⁵ M) were studied on the roots of maize (Zea mays) seedlings, cvs. Diamant and Sterling. The roots were grown on filter paper either on glass in trays or in large Petri dishes. The following characteristics of root growth were studied: seed germination, length of primary and seminal roots, number of seminal and lateral roots, length of branching zone, length of meristem and fully-elongated cells and the number of fully-elongated cells along the daily length increment. 10^-2 M lead nitrate exerted a clear toxic effect on root elongation just after radicle emergence; its influence on shoot growth was weak. However 10^-2 M Pb solution did not affect either radicle emergence itself or seminal root emergence, which can be explained by the impermeability of seed testa to lead salt. The inhibitory effect of 10^-3 M lead nitrate appeared a day later and was not as toxic: the growth of primary and seminal roots proceeded at lower rate due to a partial inhibition of cell division and cell elongation in them. 10^-3 M lead nitrate modified the root system morphology: it exerted no effect on the emergence of lateral roots and their number, but induced a more compact distribution of lateral roots along a shorter branching zone due to a reduced length of mature cells in the primary root. As a result of the more prominent inhibition of primary root growth, a shorter branching zone with more compactly located lateral roots occupied a position much closer to the root tip than in roots grown without the influence of lead.     

Acetylcholine promotes the emergence and elongation of lateral roots of Raphanus sativus

Radish (Raphanus sativus L.) was grown on four layers of paper towel moistened with distilled water with and without acetylcholine (ACh) for five days in the dark after sowing. ACh at 1 nM promoted the growth (emergence and elongation) of lateral roots of radish plants, but had no effect on the stems and main roots. Moreover, ACh enhanced the dry weight of roots [main (primary) + lateral roots]. Neostigmine, an inhibitor of acetylcholinesterase (AChE) also promoted the emergence and elongation of lateral roots, and atropine, a competitive inhibitor of ACh receptor, suppressed the emergence and elongation. ACh promoted the activities of glyceraldehyde-3-phosephate dehydrogenase (G-3-PD), nicotinamide adenine dinucleotide-specific isocitrate dehydrogenase (NAD-ICDH), succinate dehydrogenase (SDH) and cytochrome-c oxidase (Cyt-c OD) in seedlings. Moreover, ACh suppressed the activity of AChE and increased the amount of proteins and pyridine nucleotides (NAD and NADH) in the roots of the seedlings. It also increased the activities of NAD-forming enzymes [NAD synthetase and ATP-nicotinamide mononucleotide (ATP-NMN) adenyltransferase], and enhanced the amount of DNA in the roots of the seedlings. The relationship between ACh and the emergence and growth of lateral roots was discussed from a biochemical viewpoint.Key words: acetylcholine, acetylcholine esterase, DNA, lateral root, NAD, NAD-forming enzyme, protein, Raphanus sativus     

Effect of salinisation of soil on growth, water status and general nutrient accumulation in seedlings of Delonix regia (Fabaceae)

Greenhouse experiments were conducted to assess the effect of salinisation of soil on emergence, growth, water content, proline content and mineral accumulation of seedlings of Delonix regia (Hook.) Raf. (Fabaceae). Sodium chloride (NaCl) was added to the soil and salinity was maintained at 0.3, 1.9, 3.9, 6.0 and 7.9 dS m⁻¹. A negative relationship between seedling emergence and salt concentration was obtained. Salinity caused reduction in water content and water potential of tissues (leaves, stems, tap roots and lateral roots) that resulted in internal water deficit to plants. Consequently, shoot and root elongation, leaf expansion and dry matter accumulation in leaves, stems, tap roots and lateral root tissues of seedlings significantly decreased in response to increasing concentration of salt. Proline content in tissues was very low. There were no effective mechanisms to control net uptake of Na on root plasma membrane and subsequently its transport to shoot tissues. Potassium content significantly decreased in tissues in response to salinisation of soil. This tree species is a moderate salt-tolerant glycophytic plant. Nitrogen and calcium content in tissues significantly decreased as soil salinity increased. Phosphors content in tissues exhibited a declining trend with increase in soil salinity. Changes in tissues and whole-plant accumulation pattern of other elements tested, as well as possible mechanisms for avoidance of Na toxicity in this tree species in response to salinisation, are discussed.     

Effect of salinisation of soil on growth, water status and general nutrient accumulation in seedlings .of Delonix regia (Fabaceae)

Greenhouse experiments were conducted to assess the effect of salinisation of soil on emergence, growth, water content, proline content and mineral accumulation of seedlings of Delonix regia (Hook.) Raf. (Fabaceae). Sodium chloride (NaCl) was added to the soil and salinity was maintained at 0.3, 1.9, 3.9, 6.0 and 7.9 dS m^?1. A negative relationship between seedling emergence and salt concentration was obtained. Salinity caused reduction in water content and water potential of tissues (leaves, stems, tap roots and lateral roots) that resulted in internal water deficit to plants. Consequently, shoot and root elongation, leaf expansion and dry matter accumulation in leaves, stems, tap roots and lateral root tissues of seedlings significantly decreased in response to increasing concentration of salt. Proline content in tissues was very low. There were no effective mechanisms to control net uptake of Na on root plasma membrane and subsequently its transport to shoot tissues. Potassium content significantly decreased in tissues in response to salinisation of soil. This tree species is a moderate salt-tolerant glycophytic plant. Nitrogen and calcium content in tissues significantly decreased as soil salinity increased. Phosphors content in tissues exhibited a declining trend with increase in soil salinity. Changes in tissues and whole-plant accumulation pattern of other elements tested, as well as possible mechanisms for avoidance of Na toxicity in this tree species in response to salinisation, are discussed.     

Phosphate Availability Alters Lateral Root Anatomy and Root Architecture of Fraxinus mandshurica Rupr. Seedlings

Plants have evolved some mechanisms to maximize the efficiency of phosphorus acquisition.Changes in root architecture are one such mechanism. When Fraxinus mandshurica Rupr. seedlings were grown under conditions of low phosphorus availability, the length of cells in the meristem zone of the lateral roots was longer, but the length of cells in the elongation and mature zones of the lateral roots was shorter,compared with seedlings grown under conditions of high phosphorus availability. The elongation rates of primary roots increased as phosphorus availability increased, but the elongation rates of the branched zones of the primary roots decreased. The number of lateral root primordia and the length of the lateral roots decreased as phosphorus availability increased. The topological index (altitude slope) decreased as phosphorus availability increased, suggesting that root architecture tended to be herringbone-like when seedlings were grown under conditions of low phosphate availability. Herringbone-like root systems exploit nutrients more efficiently, but they have higher construction costs than root systems with a branching pattern.     

Maximum axial growth pressures of the lateral roots of pea and eucalypt

Although lateral roots may contribute significantly towards growth and nourishment of plants, the mechanics of their elongation behaviour in strong soils is not well known. The aim of this study is to report maximum axial growth pressures (p) and maximum elongation rates (E) of the lateral roots of an annual herbaceous plant (pea) and a woody perennial (eucalypt). As such measurements have not been reported previously, measurements of P and E for lateral roots were compared with the primary roots of pea for which reports are widespread. Values of P were estimated from the measured maximum values of axial force and root diameter on single, intact roots of seedlings in the laboratory. Additional measurements of both P and E were made for the lateral roots of pea when the growth of the remaining root axes was stopped (with removal of tips) to determine the overall effects of root-growth-inhibition on P and E of single roots.Values of P and E for lateral roots of pea were significantly greater than those for the lateral roots of eucalypt. Although root diameter for the primary roots of pea were similar to those for the lateral roots of eucalypt, the former exerted nearly twice as much pressure as the latter. The lateral roots of pea elongated significantly slower than the primary roots; however, P of lateral roots was significantly lower than the primary roots when elongation of all other roots was inhibited during the measurements. Production and/or development of lateral roots increased when elongation of the remaining roots (both primary and lateral roots) of pea seedlings was restricted due to the removal of root tips and exposure of one of the lateral roots to high strength. In general, maximum axial force exerted by primary and lateral roots was similar for roots of <1 mm diameter. However, primary roots exerted greater maximum axial force than the lateral roots when root diameter was >1 mm. As axial pressure of lateral roots was independent of root diameter, thickening of root tips is less likely to assist penetration of lateral roots in strong soils.     

Reactive oxygen species localization in roots of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> seedlings grown under phosphate deficiency

Arabidopsis plants responding to phosphorus (P) deficiency increase lateral root formation and reduce primary root elongation. In addition the number and length of root hairs increases in response to P deficiency. Here we studied the patterns of radical oxygen species (ROS) in the roots of Arabidopsis seedlings cultured on media supplemented with high or low P concentration. We found that P availability affected ROS distribution in the apical part of roots. If plants were grown on high P medium, ROS were located in the root elongation zone and quiescent centre. At low P ROS were absent in the elongation zone, however, their synthesis was detected in the primary root meristem. The proximal part of roots was characterized by ROS production in the lateral root primordia and in elongation zones of young lateral roots irrespective of P concentration in the medium. On the other hand, plants grown at high or low P differed in the pattern of ROS distribution in older lateral roots. At high P, the elongation zone was the primary site of ROS production. At low P, ROS were not detected in the elongation zone. However, they were present in the proximal part of the lateral root meristem. These results suggest that P deficiency affects ROS distribution in distal parts of Arabidopsis roots. Under P-sufficiency ROS maximum was observed in the elongation zone, under low P, ROS were not synthesized in this segment of the root, however, they were detected in the apical root meristem.     

Limitations of the use of plant material grown in Petri dishes for physiological experiments

Roots of plants growing "aeroponically" (AP) on moistened filter paper in Petri dishes for a few days are fairly often used for physiological experiments (e.g. measurement of root growth), for ion or herbicide uptake tests, before the establishment of hydroponic or aseptic cultures although their hormonal status is markedly different from that of the hydroponic (HP) control. On the 4th day of germination the ethylene production of cucumber (Cucumis sativus L. cv. Budai csemege) roots growing in AP under controlled conditions increased considerably and exhibited a maximum curve, HP roots evolved ethylene much more constantly. The morphological changes in AP roots (e.g. inhibited elongation and swelling of primary roots, and increased formation of root hairs), resembling those caused by exogenously applied ethylene, can be prevented with 10(-5) M Ag+, an inhibitor of ethylene action. In roots of one-week-old AP seedlings, the amount of an acidic inhibitor, which as judged from the Rf values is likely to be abscisic acid (ABA), is about twice as high as in HP seedlings. An elevated ethylene or ABA level of AP roots may result in a reduced elongation of the primary roots. Counteraction of this inhibition by Ag+ suggests that the effect of ethylene is the primary event in the reduction of root length. When using plant material grown in Petri dishes the possibility of similar changes in hormonal status of the roots must be taken into consideration.     

Phosphate Availability Alters Lateral Root Anatomy and Root Architecture of Fraxinus mandshurica Rupr. Seedlings

Plants have evolved some mechanisms to maximize the efficiency of phosphorus acquisition. Changes in root architecture are one such mechanism. When Fraxinus mandshurica Rupr. seedlings were grown under conditions of low phosphorus availability, the length of cells in the meristem zone of the lateral roots was longer, but the length of cells in the elongation and mature zones of the lateral roots was shorter,compared with seedlings grown under conditions of high phosphorus availability. The elongation rates of primary roots increased as phosphorus availability increased, but the elongation rates of the branched zones of the primary roots decreased. The number of lateral root primordia and the length of the lateral roots decreased as phosphorus availability increased. The topological index (altitude slope) decreased as phosphorus availability increased, suggesting that root architecture tended to be herringbone-like when seedlings were grown under conditions of low phosphate availability. Herringbone-like root systems exploit nutrients more efficiently, but they have higher construction costs than root systems with a branching pattem.     

Suberin lamella development in maize seedling roots grown in aerated and stagnant conditions

Hypoxia can stimulate the development of a suberized exodermis in aquatic plants; however, its influence on this aspect of terrestrial root development is sparsely documented. To determine the effects of hypoxia on maize (Zea mays cv. Seneca Horizon) roots, seedlings were grown in vermiculite (VERM), aerated hydroponics (AER), stagnant hydroponics with agar (STAG), or aerated hydroponics with agar (AERAG). The endo- and exodermis were examined for wall modifications. Lateral root emergence and aerenchyma formation were documented qualitatively. The endodermal Casparian band formation was unaffected by treatment. Endodermal and exodermal suberin lamella formation was earliest and most extensive in VERM. Suberization, especially in the exodermis of aerated treatments, was depressed in all hydroponic media. In comparison with AER, STAG exodermal lamellae were increased, but endodermal lamellae were decreased. Since the suberized exodermis forms a barrier to radial oxygen loss from roots to the medium, its stimulation in STAG roots (which also developed extensive aerenchyma) would help retain oxygen in the root. The reduction of endodermal lamellae should facilitate oxygen diffusion into the stele. Clearly, the response to environmental conditions is variable within individual cortical cell layers. Additionally, the observed patterns of lamellae, aerenchyma and lateral root development indicate a tight radial co-ordination of root development.     

Chilling Sensitivity of Photosynthetic Oil-Seedlings

Seed germination and seedling emergence are particularly sensitiveto low temperatures in oil-seed genera in which lipids are themain seed storage material, e g cotton, corn, cucumber, andwatermelon Growth of photosynthetic seedlings of oil-seed generais also significantly affected by suboptimal temperatures. Aphotosynthetic seedling growth system used to demonstrate suboptimaltemperature stress effects in cotton was modified to monitorthe effects of temperatures from 10 to 35 ? upon the roots andaerial portions of cucumber, cantaloupe, and watermelon seedlings.Non-destructive measurements of fresh weights and the relativewater contents of the roots and shoots of seedlings grown at30 ? and at non-optimal temperatures, as well as chilled seedlingsreturned to 30 ? for one day, indicated significant intra-genencdifferences in temperature sensitivity and relative root andshoot water status of both the chilled seedlings and those recoveringfrom thermal stress The capacity to return to normal water statusand resume growth differed between and within species and wascorrelated with recommendations for chilling tolerance Key words: Temperature, water stress, photosynthetic seedling     

Changes in hormonal balance and meristematic activity in primary root tips on the slowly rotating clinostat and their effect on the development of the rapeseed root system

The morphometry of the root system, the meristematic activity and the level of indole-3-acetic acid (IAA), abscisic acid (ABA) and zeatin in the primary root tips of rapeseed seedlings were analyzed as functions of time on a slowly rotating clinostat (1 rpm) or in the vertical controls (1 rpm). The fresh weight of the root system was 30% higher throughout the growth period (25 days) in clinorotated seedlings. Morphometric analysis showed that the increase in biomass on the clinostat was due to greater primary root growth, earlier initiation and greater elongation of the secondary roots, which could be observed even in 5-day-old seedlings. However, after 15 days, the growth of the primary root slowed on the clinostat, whereas secondary roots still grew faster in clinorotated plants than in the controls. At this time, the secondary roots began to be initiated closer to the root tip on the clinostat than in the control. Analysis of the meristematic activity and determination of the levels in IAA, ABA and zeatin in the primary root tips demonstrated that after 5 days on the clinostat, the increased length of the primary root could be the consequence of higher meristematic activity and coincided with an increase in both IAA and ABA concentrations. After 15 days on the clinostat, a marked increase in IAA, ABA and zeatin, which probably reached supraoptimal levels, seems to cause a progressive disturbance of the meristematic cells, inducing a decrease of primary root growth between 15 and 25 days. These modifications in the hormonal balance and the perturbation of the meristematic activity on the clinostat were followed by a loss of apical dominance, which was responsible for the early initiation of secondary roots, the greater elongation of the root system and the emergence of the lateral roots near the tip of the primary root.     

Water deficit accelerates determinate developmental program of the primary root and does not affect lateral root initiation in a Sonoran Desert cactus (Pachycereus pringlei, Cactaceae)

Determinate root growth is an important adaptation feature for seedling establishment in some Cactaceae. We show that seedlings of Pachycereus pringlei have primary roots with a stable determinate developmental program. How water stress affects determinate root growth and lateral root development has not been studied. Here we address this question. Root growth was analyzed in plants growing in vitro under well-watered and water-deficient (created by polyethylene glycol) growth conditions. Under severe water stress roots terminated their growth earlier and the rate of growth was significantly decreased as a result of inhibition of both cell elongation and cell production. Under severe water stress the number of lateral roots and primordia per millimeter of primary root was 1.5-1.7 times greater than under well-watered conditions; however, the total number of lateral roots and primordia was the same under all conditions. Lateral roots resembled root spurs found in some Opuntioideae. Analysis of the dynamics of meristem exhaustion indicated that initial-cell activities are required for the maintenance of proliferation before meristem exhaustion. We conclude that lateral root formation is a stable developmental process resistant to severe water stress and that water stress accelerates the determinate developmental program of the primary root. Both of these features appear to be important for successful seedling establishment in a desert.     

A role for redox factors in shaping root architecture under phosphorus deficiency

The developmental response of the Arabidopsis root system to low phosphorus (P) availability involves the reduction in primary root elongation accompanied by the formation of numerous lateral roots. We studied the roles of selected redox metabolites, namely, radical oxygen species (ROS) and ascorbic acid (ASC) in the regulation of root system architecture by different P availability. Rapidly growing roots of plants grown on P-sufficient medium synthesize ROS in root elongation zone and quiescent centre. We have demonstrated that the arrest of root elongation at low P medium coincides with the disappearance of ROS from the elongation zone. P-starvation resulted in a decrease in ascorbic acid level in roots. This correlated with a decrease in cell division activity. On the other hand, feeding P-deficient plants with ASC, stimulated mitotic activity in the primary root meristem and partly reversed the inhibition of root growth imposed by low P conditions. In this paper, we discuss the idea of the involvement of redox agents in the regulation of root system architecture under low P availability.Key words: ascorbic acid, phosphate deficiency, primary root, radical oxygen species, root growth, root system architecture     

The Influence of Steep Slopes on Root System Development

Mechanical forces active on steep slopes tend to overturn plants, which respond by developing a specific asymmetrical architecture in the root system. This asymmetric architecture is the consequence of preferential lateral root emergence and elongation in the up-slope and down-slope directions. Root systems show a normal symmetrical architecture when the same species is grown on plane soil. The asymmetrical root architecture on steep slopes seems to increase the plant's stability by modifying the distribution of mechanical forces into the soil. This hypothesis is supported by the observation that lateral roots developing in the up-slope or down-slope directions present considerable anatomical modifications in shape and tissue-organization compared with lateral roots from plants growing on plane soil.     

The Influence of Steep Slopes on Root System Development

Mechanical forces active on steep slopes tend to overturn plants, which respond by developing a specific asymmetrical architecture in the root system. This asymmetric architecture is the consequence of preferential lateral root emergence and elongation in the up-slope and down-slope directions. Root systems show a normal symmetrical architecture when the same species is grown on plane soil. The asymmetrical root architecture on steep slopes seems to increase the plant's stability by modifying the distribution of mechanical forces into the soil. This hypothesis is supported by the observation that lateral roots developing in the up-slope or down-slope directions present considerable anatomical modifications in shape and tissue-organization compared with lateral roots from plants growing on plane soil.     

Effects of sodium tungstate on the ultrastructure and growth of pea (Pisum sativum) and cotton (Gossypium hirsutum) seedlings

Pea (Pisum sativum L. cv. Onmard) and cotton (Gossypium hirsutum L. cv. Campo) seedlings were treated with two concentrations (200 and 500 mg/l) of sodium tungstate (Na₂WO₄) and the developmental effects were investigated. Tungstate retarded seedling growth rate and stopped root elongation in both species. Seedling growth recovered when tungstate was removed, but primary roots continued to be stunted, while lateral root initiation and growth were stimulated. Tungstate induced premature vacuolation in cells of the root apical meristem, with vacuoles having an unusual semi-circular or cap-like shape around the nucleus. In control roots, the nuclei were spherical with prominent nucleoli bearing several randomly distributed fibrillar centres. In the tungstate-treated cells nuclei contained spherical nucleoli with a big nucleolar vacuole. Occasionally, cytoplasmic components, such as mitochondria, were entrapped in the nucleoplasm of interphasic cells of the treated roots. In these roots, most cell plates were fused to only one lateral parental wall suggesting a non-uniform centrifugal extension. The vesicles in these cell plates were dark and fused to each other at a much lower rate than in the dividing cells of the untreated seedlings. Phragmoplast and cortical microtubules were abundant in the untreated cells, but scarcely detected in the treated ones. All these observations are consistent with the view that tungstate causes considerable toxic effects to pea and cotton seedlings.     

一种简单有效的非损伤观测根瘤和根系形态的方法

实时观测根瘤及根系形态对于豆科植物研究具有重要意义, 但目前还缺乏一个便于观测根系、高效结瘤、适宜生长且经济实用的豆科植物培养体系。以蒺藜苜蓿(Medicago truncatula)为植物材料, 建立了一种可实时观测根瘤及根系形态的纸袋水培法, 并与其它常用方法进行对比。结果表明, 依赖于石英砂等固体介质栽培蒺藜苜蓿对根瘤和根系形态的实时观测造成障碍, 而水培和喷雾培养等方法的根瘤菌接种效率不高, 且不便观测侧根发育情况。采用纸袋水培法探讨了褪黑素对蒺藜苜蓿根系发育的影响, 发现褪黑素具有降低根瘤形成效率、抑制侧根伸长、增加侧根数目以及增大侧根与主根之间夹角等作用。因此, 纸袋水培法能够高效接种根瘤菌且为实时无损伤观测根瘤及根系形态提供了可能, 是一种适用于豆科植物简单有效的培养方法。     

Nitric oxide is involved in nitrate-induced inhibition of root elongation in Zea mays

BACKGROUND AND AIMS: Root growth and development are closely dependent upon nitrate supply in the growth medium. To unravel the mechanism underlying dependence of root growth on nitrate, an examination was made of whether endogenous nitric oxide (NO) is involved in nitrate-dependent growth of primary roots in maize. METHODS: Maize seedlings grown in varying concentrations of nitrate for 7 d were used to evaluate the effects on root elongation of a nitric oxide (NO) donor (sodium nitroprusside, SNP), a NO scavenger (methylene blue, MB), a nitric oxide synthase inhibitor (N(omega)-nitro-L-arginine, L-NNA), H(2)O(2), indole-3-acetic acid (IAA) and a nitric reducatse inhibitor (tungstate). The effects of these treatments on endogenous NO levels in maize root apical cells were investigated using a NO-specific fluorescent probe, 4, 5-diaminofluorescein diacetate (DAF-2DA) in association with a confocal microscopy. KEY RESULTS: Elongation of primary roots was negatively dependent on external concentrations of nitrate, and inhibition by high external nitrate was diminished when roots were treated with SNP and IAA. MB and L-NNA inhibited root elongation of plants grown in low-nitrate solution, but they had no effect on elongation of roots grown in high-nitrate solution. Tungstate inhibited root elongation grown in both low- and high-nitrate solutions. Endogenous NO levels in root apices grown in high-nitrate solution were lower than those grown in low-nitrate solution. IAA and SNP markedly enhanced endogenous NO levels in root apices grown in high nitrate, but they had no effect on endogenous NO levels in root apical cells grown in low-nitrate solution. Tungstate induced a greater increase in the endogenous NO levels in root apical cells grown in low-nitrate solution than those grown in high-nitrate solution. CONCLUSIONS: Inhibition of root elongation in maize by high external nitrate is likely to result from a reduction of nitric oxide synthase-dependent endogenous NO levels in maize root apical cells.     

The involvement of the root apex and cytokinins in the control of lateral root emergence in lettuce seedlings

Removal of the apical 3 mm of the primary root of hydroponically-grown lettuce seedlings 3 or 5 days after sowing, prevented further elongation of the root and increased both the number and total length of lateral roots. The length of the lateral zone, i.e. the distance from the base of the parent root to the lateral nearest the tip, except on one occasion, remained the same as the control in both 3 and 5 day treatments, until the length of the decapitated root (which had ceased elongating) became limiting.Zeatin applied via the roots, at a concentration range from 3 × 10^–10 M to 10^–8 M reduced tap root extension growth at all concentrations. Lateral root emergence was enhanced by low zeatin concentrations and retarded by higher ones. In general, the lateral zone length was the same in cytokinin-treated plants as in untreated controls.