CORTICAL ONTOGENY IN ROOTS. I. ZEA MAYS

Serial growth stages of young Zea mays primary roots were analyzed for patterns of ground meristem ontogeny. The number of cell layers in the cortex decreases from approximately 15 to 11 during early root growth. The cortex arises mostly by periclinal divisions in the outer portions of the ground meristem at levels 50–150 μm from the meristem tip, although some layers of outer cortex arise beyond 150 μm. The proendodermis contributes 3–5 cell layers to the cortex, but this contribution diminishes during early seedling growth as anticlinal divisions occur in the proendodermis. The relationship between the ground meristem and protoderm changes at the tip of the meristem during root elongation.     

Suberin lamellae in the hypodermis of maize (Zea mays) roots; development and factors affecting the permeability of hypodermal layers

Abstract The development of suberin lamellae in the hypodermis of Zea mays cv. LG 11 was observed by electron microscopy and the presence of suberin inferred from autoliuorescence and by Sudan black B staining in nodal (adventitious) and primary (seminal) root axes. Suberin lamellae were evident at a distance of 30–50 mm from the tip of roots growing at 20°C and became more prominent with distance from the tip. Both oxygen deficiency and growth at 13°C produced shorter roots in which the hypodermis was suberized closer to the root tip. There were no suberin lamellae in epidermal cells or cortical collenchyma adjacent to the hypodermis. Plasmodesmata were not occluded by the suberin lamellae: there were twice as many of them in the inner tangential hypodermal wall (1,14 μn^?2) as in the junction between the epidermis and hypodermis (0.54 μm^?2). Water uptake by seminal axes (measured by micropotometry) was greater at distances more than 100 mm from the root lip than in the apical zone where the hypodermis was unsuberized. In the more mature zones of roots grown at 13°C rates of water uptake were greater than in roots grown at 20°C even though hypodermal suberization was more marked. Sleeves of epidermal/hypodermal cells (plus some accessory collenchyma) were isolated from the basal 60 mm of nodal axes by enzymatic digestion (drisclase). The roots were either kept totally immersed in culture solution or had the basal 50 mm exposed to moist air above the solution surface. In both treatments the permeabilities to tritiated water and ₈₆Rb were low (circa 10^?5mms^?1) in sleeves isolated from the extreme base. In roots grown totally immersed, however, the permeability of sleeves increased 10 to 50-fold over a distance of 40 mm. In roots exposed to moist air the permeability remained at a low level until the point where the root entered the culture solution and then increased rapidly (> 50-fold in a distance of 8 mm). Growth of roots in oxygen depleted (5% O₂) solutions promoted the development of extensive cortical aerenchymas. These developments were not associated with any reduction in permeability of sleeves isolated from the basal 40 mm of the axis. It was concluded that the presence of suberin lamellae in hypodermal walls does not necessarily indicate low permeability of cells or tissues to water or solutes. The properties of the walls (lamellae?) can be greatly changed by exposure to moist air, perhaps due to increased oxygen availability.     

Metabolism and binding of C-maleic hydrazide

Maleic hydrazide (MH) is taken up by corn and pea seedling roots and bound to some material which is insoluble in 80% ethanol or 5% trichloroacetic acid. ¹⁴C-MH is stable metabolically; chromatography of the 80% ethanol-soluble ¹⁴C from treated corn roots and tobacco pith gives no indication of degradation. Very little ¹⁴C-MH is bound in the zone of cell division (where MH acts to inhibit root elongation) or even in the region of cell enlargement in corn roots and most is bound 1 or more centimeters behind the tip. Likewise, very little MH is bound in corn coleoptile and tobacco pith sections. About 90% of ¹⁴C-MH bound in corn roots is associated with large particles which may be cell wall fragments. The binding is blocked by azide and dinitrophenol, indicating a requirement for metabolic energy; however, inhibitors of protein synthesis (chloramphenicol, puromycin, cycloheximide) and DNA synthesis (fluorodeoxyuridine) do not inhibit binding. Only very small amounts of MH are bound in root homogenates, providing further evidence that the binding process is active. Once the MH is bound in the roots, the complex is stable for at least 1 week. Treatment with 2-aminoethanol releases MH.     

Laser ablation ICP-MS reveals patterns of copper differing from zinc in growth zones of cucumber roots

Laser ablation coupled with inductively coupled plasma-mass spectrometry was used to find Cu and Zn concentration in surface tissue along a longitudinal developmental gradient with meristem, rapidly elongating tissue, and nongrowing tissue in a model system of seedling roots of Cucumis sativus L. (cucumber). Tissue metal accumulation was determined for roots of seedlings growing on cellulosic germination paper treated with nutrient solution (controls), and also treated with concentrations of Zn (40 ppm) and Cu (10 ppm) that reduced growth. Cu content of all roots is highest at the apex and falls sharply to lower values by 2 mm from the root tip. In contrast, at moderate Zn availability (0.07 ppm), Zn content rises from the apex to 2 mm then falls throughout the remainder of the growth zone. At high external Zn the spatial pattern resembles that of Cu. Cucumber root growth zones accumulate more of each metal with higher external availability. Metal deposition rates were calculated using a continuity equation with data on local metal content and growth velocity. Deposition rates of both metals are generally highest in the rapidly elongating region, 1.5–3.5 mm, even where metal concentration is decreasing with position and root age and even when the accumulation is inhibitory to growth.     

Aerenchyma formation and associated oxygen movement in seminal and nodal roots of wheat

Abstract The present paper describes the effects of growth of roots of wheat (Triticum aestivum cv. Gamenya) in hypoxic nutrient solutions on acrenchyma formation and O₂ movement from shoots to roots. Two types of roots were investigated: (1) seminal roots of 4–7-d-old seedlings, and (2) seminal and nodal roots of 10–28-d-old plants. Gas-filled porosity of seminal and nodal roots increased from 3 to 12% and from 5–7 to 11–15%, respectively, when the roots emerged in stagnant or N₂-flushed solutions (0.003 mol m ^?3 O₂) compared with growth in continuously acrated solutions (0.26 mol m ^?3 O₂). However, neither root type increased in porosity when they were longer than 100–200 mm at the start of the exposure to these stagnant or N₂-flushed treatments. A vernier microscope and cylindrical platinum-electrode were used to examine the relationship between root extension and transport of O₂ from shoots to roots via the gas spaces. Measurements were made when the roots were in an anoxic medium and were dependent solely on O₂ supplied from the shoots. For seminal roots of 5–7-d-old seedlings raised in stagnant solutions (90–100 mm), internal O₂ transport was sufficient to support a rate of root elongation in the O₂-free medium of between 0.03 and 0.17 mm h^?1. When the O₂ pressure around the shoots was increased from 20 to 100 kPa O₂, the O₂ concentrations at the walls of the expanding zone (2–7 mm from the tip) of these roots increased from 0.006 mol m^?3 to between 0.04 and 0.26 mol m^?3, and the rate of root extension increased five-fold. Oxygen transport to roots grown continuously in acrated solutions was considerably less than for roots raised in stagnant solutions; this difference was greater for seminal than for nodal roots. When the acrated seminal roots were longer than 100 mm and transferred to an O₂-free root medium, O₂ concentration became zero at the root tip causing elongation to cease. After 24 h of anoxia, none of these roots were able to resume elongation following a return to acrated solutions.     

STUDIES ON THE GERMINATION OF SEEDS OF THE ROOT PARASITES,ALECTRA VOGELII AND STRIGA GESNERIOIDES. I. ANATOMICAL CHANGES IN THE EMBRYOS

Anatomical changes in the radicle and shoot meristems of embryos of germinating seeds of the obligate root parasites, Alectra vogelii and Striga gesnerioides were studied. When germination of seeds was stimulated by cowpea (Vigna unguiculata) root exudate, growth occurred mainly in the radicular pole of embryos and minimally in the plumular pole, resulting in seedlings with elongated radicles. Maximum radicle elongations of about 3 mm in A. vogelii and 2 mm in S. gesnerioides were recorded during a period of 8 and 11 days, respectively. Analysis of the radicular tip during the course of seed germination revealed that the activity of the meristematic tissue progressively decreased until it completely disappeared. When germinated seeds were cultured on nutrient agar media, the radicle meristem of A. vogelii continued to grow producing a normal root with a root cap. On the other hand, the radicles of cultured S. gesnerioides seeds elongated only slightly before meristematic activity ceased. During continued growth of seedlings of both species on agar media, lateral roots whose tips had typical angiosperm root topography, were initiated from the radicle.     

Cell Division Heterogeneity in the Primary Meristems of Pisum sativum Root Tips after Excision

Root tips (1.0 cm) were excised, transferred to culture medium,sampled at intervals, embedded in paraffin and sectioned. Themitotic index in each primary meristem in the 2.0 mm tips wasreduced immediately after excision. After 8–12 h mitosisresumed. The position of mitotic figures in the protoderm, groundmeristem, procambium, and root cap was determined. The greatestnumber of mitotic figures occurred in the procambium, and uponexcision the smallest percent reduction also occurred in thisregion. The ground meristem showed the greatest reduction inmitotic figures. After excision, the relative contribution tothe total number of mitotic figures increased in the procambiumtissue and decreased in the ground meristem. Cell division inall primary meristems of the root tip is reduced after tip excision,but the ground meristem (cortical region) is the primary targettissue.     

Relationships among root branch order, carbon, and nitrogen in four temperate species

The objective of this study was to examine how root length, diameter, specific root length, and root carbon and nitrogen concentrations were related to root branching patterns. The branching root systems of two temperate tree species, Acer saccharum Marsh. and Fraxinus americana L., and two perennial herbs from horizontal rhizomes, Hydrophyllum canadense L. and Viola pubescens Ait., were quantified by dissecting entire root systems collected from the understory of an A. saccharum-Fagus grandifolia Ehrh. forest. The root systems of each species grew according to a simple branching process, with laterals emerging from the main roots some distance behind the tip. Root systems normally consisted of only 4–6 branches (orders). Root diameter, length, and number of branches declined with increasing order and there were significant differences among species. Specific root length increased with order in all species. Nitrogen concentration increased with order in the trees, but remained constant in the perennial herbs. More than 75% of the cumulative root length of tree seedling root systems was accounted for by short (2–10 mm) lateral roots almost always <0.3 mm in diameter. Simple assumptions suggest that many tree roots normally considered part of the dynamic fine-root pool (e.g., all roots <2.0 mm in diameter) are too large to exhibit rapid rates of production and mortality. The smallest tree roots may be the least expensive to construct but the most expensive to maintain based on an increase in N concentration with order. Received: 25 November 1996 / Accepted: 27 March 1997     

A model for dynamics of cell division cycle in onion roots

Summary The study of the cell division cycle by means of caffeine labelling inAllium roots, at 15° C, employing intact root and decapitated roots at several levels (0.5, 1.0, 1.5, 2.0, and 2.5 mm) has shown that the number of cycles developed by the cells is constant at each meristem level. This number and the durations of the cycles are not affected by the decapitation. It is suggested that the cell cycle is controlled in the meristematic cells by an intracellular programme which would be developed throughout the meristem.However, the larger the region decapitated is, the more decreases the growth rate of the roots. The removal of the root cap (about 0.5 mm) did not modify the rate of root growth, although it blocked the geotropic response. The quiescent center is proposed as a source of auxin controlling cell elongation.     

DISTRIBUTION AND RELATIONSHIP OF CELL DIVISION AND MATURATION EVENTS IN PISUM SATIVUM (FABACEAE) SEEDLING ROOTS

Pea roots have open apical organization, where discrete initial cells do not exist. Differentiation of all tissues occurs in cylinders and vascular sectors that blend gradually with each other. This study reports the distribution of dividing cells and their relationship to maturation events in the 2 mm root tip, and in the 8–10 and 18–20 mm segments. Up to 200 μm from the root body/cap junction, cell division is uniformly distributed throughout all meristem regions. By 350 to 500 μ, xylem tracheary elements and cells of the pith parenchyma and middle cortex have stopped dividing. At this level cell division is almost entirely restricted to two cylinders, one composed of the inner root cap, the epidermis, and the outer cortex (outer cortex cylinder) and another composed of cells of the inner cortex, the pericycle and vascular tissue (inner cortex cylinder). When the protophloem matures, all cells in the phloem sector of the inner cortex cylinder, including the 1 layered pericycle, the endodermis and the phloem parenchyma, stop dividing. The 3–4 layered pericycle in the xylem sectors continues dividing until about 10 mm from the body/cap junction following the maturation of the protoxylem tracheary elements.     

The improved Allium/Vicia root tip micronucleus assay for clastogenicity of environmental pollutants

The meristematic mitotic cells of plant roots are appropriate and efficient cytogenetic materials for the detection of clastogenicity of environmental pollutants, especially for in situ monitoring of water contaminants. Among several cytological endpoints in these fast dividing cells, such as chromosome/chromatid aberrations, sister-chromatid exchanges and micronuclei, the most effective and simplest indicator of cytological damage is micronucleus formation. Although the Allium cepa and Vicia faba root meristem micronucleus assays (Allium/Vicia root MCN) have been used in clastogenicity studies about 12 times by various authors in the last 25 years, there is no report on the comparison of the efficiency of these two plant systems and in different cell populations (meristem and F₁) of the root tip as well as under adequate recovery duration. In order to maximize the efficiency of these bioassays, the current study was designed to compare the Allium and the Vicia root MCN assays on the basis if chromosome length, peak sensitivity of the mitotic cells, and the regions of the root tip where the MCN are formed. The total length of the 2n complement of Allium chromosomes is 14.4 μm and the total length of the 2n complement of Vicia is 9.32 μm. The peak sensitivity determined by serial fixation at 12-h intervals after 100 R of X-irradiation is 44 h. The slope of the X-ray dose-response curve of Allium roots derived from the meristematic regions was lower than that derived from cells in the F₁ region. Higher efficiency was also demonstrated when the MCN frequencies were scored from the F₁ cells in both Allium and Vicia treated with formaldehyde (FA), mitonycin C (MMC), and maleic hydrazide (MH). The results indicated that scoring of MCN frequencies from the F₁ cell region of the root tip was more efficient than scoring from the meristematic region. The X-ray linear regression dose-response curves were established in both Allium and Vicia cell systems and the coefficients of correlations, slope values were used to verify the reliability and efficiency of these two plant cell systems. Based on the dose-response slope value of 0.894 for Allium and 0.643 for Vicia, the Allium root MCN was a more efficient test system. The greater sensitivity of the Allium roots is probably due to the greater total length of the diploid complement and the higher number of metacentric chromosomes. The Allium/Vicia root MCN test system was applied to determine the clastogenicity of saccharin (SC) and wastewater from Rio Queretaro and the Arena canal in the city of Queretaro, Mexico. The minimum effective dose (MED) is 10 R for X-rays, 50 mM for FA, 2.2 mM for MMC, 0.01 mM for MH, and 40 ppm for SC.     

ORIGIN AND DEVELOPMENT OF LATERAL ROOTS IN TYPHA GLAUCA

Lateral roots of Typha glauca arose from the pericycle of the parent adventitious root. Periclinal divisions of the pericycle gave rise to two layers; the outermost initially produced the ground meristem and protoderm, and the innermost produced the procambium. The immature endodermis of the parent root contributed to the early stages of the root tip as an endodermal covering. Prior to emergence, the ground meristem/protoderm produced cells into the endodermal covering. After emergence, the endodermal covering was replaced by a calyptrogen, which was derived from the ground meristem/protoderm and which, in turn, formed the rootcap. A typical monocotyledonous three-tiered meristem was then produced. An outer ground meristem also arose before emergence to form a hypodermis in many lateral roots; in these, crystalliferous cell production began in midcortex cells before emergence, and a small aerenchyma developed in their cortices. The rootcap columella stored small amounts of starch shortly after emergence. Lateral roots of T. glauca were smaller than their parental adventitious roots; they normally had only two to six poles of xylem and phloem, and the cortex was less than six cells across. During 1–3-cm elongation, the lateral root apical meristem and mature regions narrowed, stored starch disappeared, fewer crystals formed, aerenchyma production ceased, and the roots stopped elongation.     

Cation-exchange capacity and pectin gradients in leek root segments

Summary Cation-exchange capacity (C.E.C.), pectin, respiration, and nitrogen content have been measured in successive segments of leek roots over a distance of 140 mm from the root tip. All are highest at the root tip and decrease with increasing distance from the root tip.     

玉米初生根向水性诱导优化试验研究

为了研究湿度梯度对根系向水性反应的影响,采用Takahashi and Scott于1993年创建的方法,设置以下3个试验:1)向水性诱导物不同倾斜角试验;2)根系距向水性诱导物不同距离试验;3)根尖距底部饱和K2CO3溶液不同距离试验。同时,还研究了根长和根系延伸速率对根系向水性弯曲的影响。结果表明,用饱和K2CO3溶液控制湿度时根系的向水性弯曲度明显大于纯水。随着诱导物倾斜角的增大,向水性弯曲增强。与距诱导物3 mm和6 mm相比,根系直接接触诱导物时表现出最大的向水性反应。与根尖距底部盐溶液6 cm相比,相距4 cm时向水性弯曲度增大,这些与根尖周围的湿度梯度增大有关。当根长为1.0、1.5、2.0、2.5、3.0 cm时,短根比长根表现出更大的向水性反应,这可能与其较慢的延伸速率为根系对湿度梯度的反应提供了更充足的时间有关。为了验证这个假说,用相同长度的根系、通过控制不同温度进行试验,结果表明根系的向水性弯曲随温度升高而降低。可见,玉米初生根的向水性反应受环境和根系发育阶段两方面影响。当根系相距诱导物较近、根系周围的湿度梯度较大时,根系向水性反应更强。而且,具有较小延伸速率根系的向水性反应更大。考虑到干旱条件下根系伸长慢、且土壤中湿度梯度大,因而可以认为干旱条件下根系的向水性生长在玉米吸收水分中有重要作用。同时,对根系向水性诱导方法的优化有助于其生理机制的进一步研究。     

Changes in plant growth processes under microgravity conditions simulated by a three-dimensional clinostat

We developed a three-dimensional (3-D) clinostat to simulate a microgravity environment and studied the changes in plant growth processes under this condition. The rate of germination of cress (Lepidium sativum), maize (Zea mays), rice (Oryza sativa), pea (Pisum sativum), or azuki bean (Vigna angularis) was not affected on the clinostat. The clinostat rotation did not influence the growth rate of their roots or shoots, except for a slight promotion of growth in azuki roots and epicotyls. On the contrary, the direction of growth of plant organs clearly changed on the 3-D clinostat. On the surface of the earth, roots grow downward while shoots upward in parallel to the gravity vector. On the 3-D clinostat, roots of cress elongated along the direction of the tip of root primordia after having changed the direction continuously. Rice roots also grew parallel to the direction of the tip of root primordia. On the other hand, roots of maize, pea, and azuki bean grew in a random fashion. The direction of growth of shoots was more controlled even on the 3-D clinostat. In a front view of embryos, shoots grew mostly along the direction of the tip of primordia. In a side view, rice coleoptiles showed an adaxial (toward the caryopsis) while coleoptiles of maize and epicotyls of pea and azuki bean an abaxial curvature. The curvature of shoots became larger with their growth. Such an autotropism may have an important role in regulation of life cycle of higher plants under a microgravity environment.     

Effect of Soybean Tip Removal on Penetration and Development of Heterodera glycines

On a few occasions, soybeans with broken root tips were included in tests to evaluate resistance to Heterodera glycines. Although females developed on these plants, the numbers tended to be lower than on similarly treated intact roots. To test the possibility that removal of the root meristem affected nematode development, a culture system using pruned soybeans was devised that permitted access to the roots without disturbing the plants. Treatments included removal of 2 mm of root tip at various times ranging from 24 hours before to 10 days after inoculation, or roots left intact. In each experiment, all roots were inoculated at the same time with equal numbers of freshly hatched second-stage juveniles of Heterodera glycines. No differences in nematode development were detected in plants with root tips removed after inoculation compared to the control. When tips were removed at or before inoculation, fewer juveniles entered roots and relatively fewer nematodes developed. Penetration levels and development correlated with root tip removal such that progressively fewer nematodes entered roots and relatively greater numbers of nematodes remained undeveloped as the time interval between root tip removal and inoculation was increased.     

三峡库区马尾松根系生物量的空间分布

以三峡库区主要植被马尾松人工林为研究对象,用内径为10 cm的根钻,分别在马尾松中龄林、近熟林和成熟林内,据树干0.5、1.0、1.5 m和2.0 m处设置取样点,各样点按0-10、10-20、20-30、30-40、40-60 cm将土壤分为5个垂直层次,对马尾松根系的空间分布格局进行调查。结果表明:(1)三峡库区马尾松总根系生物量(0-10 mm)为中龄林(4.72 t/hm²)显著高于成熟林(2.94 t/hm²)和近熟林(2.40 t/hm²)(P<0.05)。细根(0-2 mm)生物量随年龄增加而递减,差异不显著(P>0.05);(2)马尾松3个林龄中根系生物量表现出一定的水平分布特征,但具体趋势表现各异,细根生物量最大值均出现在距离样木1.0 m处;(3)细根主要分布在土壤上层,其中47.53%-71.73%的活细根集中在0-20 cm土壤深度内,且随土层的加深,其生物量明显减少。粗根(2-10 mm)则主要分布于20-60 cm土层范围内;(4)根系直径越小,受环境变化越明显。马尾松细根生物量分布主要受土壤深度的影响,树龄和不同水平距离对细根分布格局影响不显著(P>0.05),各因素对粗根生物量的影响均未达到显著水平(P>0.05)。     

Analysis of Cell Division and Elongation Underlying the Developmental Acceleration of Root Growth in Arabidopsis thaliana

To investigate the relation between cell division and expansion in the regulation of organ growth rate, we used Arabidopsis thaliana primary roots grown vertically at 20°C with an elongation rate that increased steadily during the first 14 d after germination. We measured spatial profiles of longitudinal velocity and cell length and calculated parameters of cell expansion and division, including rates of local cell production (cells mm⁻¹ h⁻¹) and cell division (cells cell⁻¹ h⁻¹). Data were obtained for the root cortex and also for the two types of epidermal cell, trichoblasts and atrichoblasts. Accelerating root elongation was caused by an increasingly longer growth zone, while maximal strain rates remained unchanged. The enlargement of the growth zone and, hence, the accelerating root elongation rate, were accompanied by a nearly proportionally increased cell production. This increased production was caused by increasingly numerous dividing cells, whereas their rates of division remained approximately constant. Additionally, the spatial profile of cell division rate was essentially constant. The meristem was longer than generally assumed, extending well into the region where cells elongated rapidly. In the two epidermal cell types, meristem length and cell division rate were both very similar to that of cortical cells, and differences in cell length between the two epidermal cell types originated at the apex of the meristem. These results highlight the importance of controlling the number of dividing cells, both to generate tissues with different cell lengths and to regulate the rate of organ enlargement.     

Growth and curvature in seedlings of Pisum sativum and an ageotropic mutant

In an attempt to explain the influence of gravity on the behaviour of ageotropic plant organs, a pea mutant (Pisum sativum ageotropum) and normal pea (Pisum sativum cv. Sabel) were examined. The mutant has a significantly lower germination rate (large seeds: 25%, small seeds: 10%) than normal pea seeds (55%). Removal of testa increased germination dramatically, the values obtained were 63 and 89%, respectively. Immediately after imbibition the mutant from which the testa had been removed, developed more slowly than normal pea seeds; after 28 h the difference in elongation rate between the two types was reversed. When continuously stimulated geotropically in the horizontal position the elongation in the mutant is larger than in the normal pea roots kept in the same position. During a 24 h period starting 48 h after imbibition the mutant root elongated 45.0 mm while the value for the normal pea root was 11.5 mm. The course of the geotropic curvature in roots of the two types has been followed during a period of 24 h. Normal pea roots develop an asymmetry in the extreme root tip region after 30 min of horizontal stimulation. After prolonged stimulation (exceeding 2 h) the asymmetry has disappeared and the curvature distributed over the entire growth region. When roots of normal pea are stimulated continuously at various angles, the optimum angle of geotropic response is 90° with decreasing responses in the order 135° (i.e. the root tip is pointing obliquely upward) and 45°. The presumed ageotropic behaviour of the mutant has only to a certain extent been confirmed in the present study. When stimulated at 135° a slight positive curvature developed; stimulation at 90° and 45° gave a slight negative curvature.