The Relationship Between Cell Size and Cell Number and the Distance from the Root Tip to the Region of Phloem Initiation in Groundnuts (Arachis hypogea. L. var. Bukene Red)

The effects of cell size and number on the distance from theroot tip to phloem inception in groundnuts have been investigated.The roots were treated with a number of plant growth regulatorshaving a wide range of effects on the growth rate. Increasing the growth rate of roots increased the distance ofphloem initiation and lignification from the apex. In all treatments,the average distance to phloem inception varied from about 590to 885 µm behind the root apex. The average lengths of the cortical and epidermal cells, whichwere 16 and 20 µm respectively, remained constant regardlessof the distance from the root tip.     

Experimental evaluation of an efflux-influx model of C exudation by individual apical root segments

The aim of this study was to evaluate if a model describing the efflux and the influx of C through the root surface could be fitted to experimental short-term kinetics of carbon (C) exudation by individual apical root segments in maize (Zea mays L.). The efflux of C was set constant or modelled by a power function of the distance from the apex to simulate the greater release of C around the root tip commonly reported in the literature. The influx was proportional to the C concentration in the external solution to simulate the active re-uptake of exudates by the root. Plants were exposed to full light or to shade to manipulate C allocation to roots. The model with a constant efflux gave satisfactory fits to the kinetics of exudation (average R(2)=0.66). The average gross efflux was then 2.1 mug C cm(-2) root surface h(-1). The model was improved if exudation was set more intense towards the root apex (average R(2)=0.74). The estimated gross efflux decreased then from 5.2 mug C cm(-2) h(-1) at the apex to 1.8 mug C cm(-2) h(-1) for the region located 5-25 cm from the root tip. The decrease in net exudation of individual roots due to the shading of plants was weak, which may indicate that the import of C by the primary roots studied was not reduced significantly. By describing the exudation of an apical root segment of variable length and diameter, the model is a first step in linking exudation to root system architecture models and to whole plant functioning.     

Endoplasmic microtubules configure the subapical cytoplasm and are required for fast growth of Medicago truncatula root hairs

To investigate the configuration and function of microtubules (MTs) in tip-growing Medicago truncatula root hairs, we used immunocytochemistry or in vivo decoration by a GFP linked to a MT-binding domain. The two approaches gave similar results and allowed the study of MTs during hair development. Cortical MTs (CMTs) are present in all developmental stages. During the transition from bulge to a tip-growing root hair, endoplasmic MTs (EMTs) appear at the tip of the young hair and remain there until growth arrest. EMTs are a specific feature of tip-growing hairs, forming a three-dimensional array throughout the subapical cytoplasmic dense region. During growth arrest, EMTs, together with the subapical cytoplasmic dense region, progressively disappear, whereas CMTs extend further toward the tip. In full-grown root hairs, CMTs, the only remaining population of MTs, converge at the tip and their density decreases over time. Upon treatment of growing hairs with 1 microM oryzalin, EMTs disappear, but CMTs remain present. The subapical cytoplasmic dense region becomes very short, the distance nucleus tip increases, growth slows down, and the nucleus still follows the advancing tip, though at a much larger distance. Taxol has no effect on the cytoarchitecture of growing hairs; the subapical cytoplasmic dense region remains intact, the nucleus keeps its distance from the tip, but growth rate drops to the same extent as in hairs treated with 1 microM oryzalin. The role of EMTs in growing root hairs is discussed.     

Wavelike distributions of infections by an introduced and naturally occurring root pathogen along wheat roots

Previously, we showed that copiotrophic and oligotrophic bacteria fluctuate as moving waves along roots. These waves probably originate as a result of growth and death cycles at any location where a moving nutrient source passed. In this study, we placed sclerotia of Rhizoctonia solani AG8 along growing roots of wheat and showed that the proportions of root sections from which R. solani was isolated fluctuated with distance from the root tip. Similarly, proportions of root sections from which naturally occurring Pythium spp. were isolated fluctuated with distance from the root tip. Fourier analysis showed that these fluctuations constituted significant waves. Cross-correlation analyses demonstrated that there were negative correlations between R. solani infections and colony forming units of copiotrophic bacteria at the time of inoculation at the same locations on the root (lag = 0 cm), indicating that infection by R. solani could have been inhibited by these bacteria. There was a positive correlation between Pythium infections and copiotrophic bacteria at a lag of 6 cm along the roots. It therefore appears that Pythium infection took place shortly after the initial peak in copiotrophic bacteria following the passage of the root tip.     

A Correlation between Structure and Function in the Root of Zea mays

The exudation rates of fluid and potassium ions from isolatedmaize roots were determined before and after excision of certainlengths of root tip. The results of this study suggest thatexcised maize roots possess the ability to absorb potassium(and presumably chloride) ions and concomitant amounts of waterover a considerable distance (10 cm) from the tip. Moreover,the observed power of absorption of ions and water into thetranslocatory pathway decreases in passing from the tip towardsthe base of the root. Both light and electron microscope techniques were used to examinethe anatomy of primary roots similar to those used in the physiologicalexperiments. The principal observation was that the xylem vesselsnear the root tip contain membrane-bounded cytoplasm with organelles.The number of mature xylem vessels, i.e. without cytoplasm,progressively increased in transverse sections cut from 1 to10 cm from the root tip; above 10 cm from the root tip all ofthe xylem vessels were found to be completely mature. It isevident that prima facie a connexion exists between this singleaspect of root anatomy and fluid exudation from excised roots. The uptake of tritiated water by roots and its transport intoexudates was examined. These data were analysed on the assumptionthat the exchange of external labelled water with the exudatewas achieved by the fluid exudation itself; this analysis indicatedthat an operational volume, similar to that of the total xylemvolume within the root, must become labelled during the formationof the exudate.     

Nuclear dynamics during the simultaneous and sustained tip growth of multiple root hairs arising from a single root epidermal cell

Nuclear dynamics in root hairs, which depends upon the actin cytoskeleton, appears to be an important factor in root-hair tip growth. Previous evidence suggests that there is an absolute requirement for the nucleus to be a fixed distance from the growing root-hair tip for tip growth to proceed. To test this hypothesis, nuclear dynamics were examined in root-hair cells bearing multiple root hairs. The majority of root-hair cells of transgenic plants overexpressing the ROP2 GTPase (ROP2 OX) bear multiple root hairs. Simultaneous and sustained fast tip growth occurred in multiple root hairs of ROP2 OX, with the continual presence of tip-localized cytoplasm in these growing hairs. Nuclear dynamics were imaged in ROP2 OX by co-expressing a transgene encoding a nuclear localization signal (NLS)-green fluorescent protein (GFP) fusion protein. The nucleus was in continual proximity to one of the growing root-hair tips, whilst the other tip elongated at a similar rate but in the absence of the nucleus from the shank of that root hair. To test whether this phenomenon was an artefact of ROP2 overexpression, nuclear dynamics were examined in wild-type and NLS-GFP transgenic plants. Multiple root hairs on the same cell underwent simultaneous and sustained fast tip growth, with the nucleus lying deep within the shank of only one of these hairs. The nucleus was also moved into the root-hair tip during the severe root-hair tip branching which is characteristic of ROP2 OX transgenic plants. These results suggest that fast tip growth can proceed in some multiple root hairs at extreme distances from the nucleus.     

DEVELOPMENT OF THE WOODY PORTION OF THE ROOT SYSTEM OF BETULA PAPYRIFERA

Our ontogenetic analysis of paper birch root systems shows that the fate of a root tip is related to its relative primary xylem diam (PXD). Lateral root tips with an initial PXD less than about 25 % of the parent root PXD are ephemeral. Some tips having a PXD of more than about 25 % of the parent root PXD become the permanent portions of the root system, enlarging over time as they elongate. In seedling root systems, the primary root and first-formed laterals are initially about the same size, and their PXD's all enlarge with increasing distance from the stem as the tips elongate to form the initial horizontal woody framework. Permanent lateral root branches with a large relative PXD develop after root tip injury, when the root tip is forced to grow in a curve, or from other unaccountable causes. Our observations show the importance of using relative diameters when classifying roots and when applying the concept of heterorhizy to paper birch root systems. Evidence points to the existence of some form of apical dominance in roots.     

Changes in the Nucleus during Cellular Development in the Pea Seedling

Nuclei were isolated from three regions of the root and fromthe epicotyls of growing pea seedlings. As the dry mass of thenuclei was the same before and after isolation, and as the DNAcontent of the nuclei accounted for the DNA of the cells, theisolated nuclei are assumed to be a random sample of the nucleiin the intact tissues. The DNA, RNA, and protein contents percell were much greater in the epicotyl than in the root, butthe RNA and protein contents of the nucleus were similar inboth epicotyl and root. Although the amounts of DNA, RNA, andprotein per cell increased with increasing distance from theroot tip, increase in the protein content of the nucleus occurredmainly between the meristematic and expanding regions, and atthis same point nuclear RNA decreased. Nuclear volume increasedwith increasing distance from the root tip, whereas nucleardry mass remained virtually unchanged. The nucleus thereforebecomes increasingly hydrated as the cells mature. The datasuggest a changing interaction between the nucleus and the cytoplasmduring cellular development.     

Formation of appressoria by the arbuscular mycorrhizal fungus <Emphasis Type="Italic">Gigaspora margarita</Emphasis> on roots of <Emphasis Type="Italic">Allium cepa</Emphasis> is linked with root age

Spores of the arbuscular mycorrhizal fungus, Gigaspora margarita, were placed near the root tip, the middle of the root (equal distance from root base and root tip), or the root base (close to the shoot) of the first primary root of 9-day-old onion. Two weeks later, the number and position of appressoria and the appressoria with penetrating hyphae were determined in the first and the newly formed second primary roots. The total number of appressoria was not significantly different among the treatments. Inoculation near the root tip of the first primary root resulted in the formation of a large number of appressoria on the first primary root and the formation of about three times fewer appressoria on the second primary root. Inoculation near the base of the first primary root resulted in the formation of no appressoria on the first primary root, whereas many appressoria were formed on the second primary root. Our results suggest that the root age is a determinant of the appressorium formation.     

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.     

STUDIES ON THE GROWTH HORMONE OF PLANTS : VI. THE DISTRIBUTION OF THE GROWTH SUBSTANCE IN PLANT TISSUES

1. It is shown that when plant tissues are ground with water the growth substance contained therein is inactivated by the oxidizing enzymes. 2. A simple method of extraction is described which enables the quantitative determination of growth substance in such tissues. 3. The amount and distribution of growth substance in the Avena coleoptile is determined by this method, and it is shown that while the substance does not diffuse out from the lower parts of the coleoptile, it is nevertheless present in considerable amounts, the concentration decreasing steadily with the distance from the tip. 4. Growth substance is also present in considerable amounts in Avena roots, and here also its concentration decreases steadily with distance from the tip. 5. The amount of growth substance diffusing out of root tips into dextrose agar, even during long periods of time, is not greater than the amount obtainable by direct extraction. Actual production in the root tip therefore either does not take place at all, or else takes place under quite different conditions from the production in the tip of the coleoptile.     

ON THE CORRELATION OF PRIMARY ROOT LENGTH,MERISTEM SIZE AND PROTOXYLEM TRACHEARY ELEMENT POSITION IN PEA SEEDLINGS

Pea seedlings (Pisum sativum L. cv. Alaska) were darkgrown in vermiculite. Roots of various lengths were cleared, stained and measured to determine the relative meristem height (MH), width and volume and the distance to the most proximal protoxylem tracheary element (PTE). A correlation was found between root length, MH and PTE position as follows: in roots from 4–40 mm as the root elongated the MH lengthened and PTE position increased its distance from the body/cap juncture; in roots 40–80 mm MH and PTE position remained approximately constant; in longer roots (80–120 mm) MH became shorter, and PTE position closer to the tip as the root elongated. The relationship, using our measurement procedure was that for every 0.19 mm change in MH, the PTE position changed by 1 mm. This response was partially growth rate dependent since short roots (4–80 mm) grew at a constant rate and longer roots (80–140 mm) grew slower. Root manipulations and trifluralin treatment, to inhibit cell division, caused tip swelling and modulated the position of the PTE toward the root tip. The control of the spatial relationship between meristem size and maturation position is discussed.     

Seasonal variations in the spatial distribution of root tips in teak (Tectonia grandis Linn. f.) plantations in the Varanasi Forest Division,India

Summary Seasonal variations in the spatial distribution of root tips were studied in 19 and 29 year old teak plantations, located on red and alluvial soils respectively. The pattern was essentially similar at both sites, but generally the alluvial soil site exhibited a greater number of root tips. Root tips decreased with increasing distance from the tree base. Through-out most of the year the relative distribution of root tips decreased with depth; the difference between 0–10 and 10–20 cm depths was marginal, but 20–40 cm depth contained distinctly fewer root tips. At all distances a similar seasonal trend was noticed, a mid rainy season peak being followed by a steady decline until the dry summer except for an abrupt rise to a smaller peak in February after the winter rains.The root tip density was positively correlated with the 2 mm root biomass and both showed a similar bimodal annual cycle. Of three environmental variables studied, soil moisture and rainfall were significantly positively correlated with root tip densityl the relationship between soil temperature and root tip density was negative and non-significant. The combined effect of soil moisture and temperature on root tip density, evaluated by a multiple regression model, accounted for 80–95% of the variation in root tip density.     

TRICHOBLASTS IN HYDROCHARIS. I. ORIGIN. DIFFERENTIATION,DIMENSIONS AND GROWTH

In the root epidermis of Hydrocharis morsus-ranae L. distinctive trichoblasts, which later grow out as root hairs, are formed by the unequal division of protodermal (immature epidermal) cells. The trichoblast is the more proximal product of this division. Trichoblasts differ from adjacent epidermal cells in manner of growth, in size, amount of cytoplasm, degree of succinic dehydrogenase and cytochrome oxidase activity, and in the structure of their plastids. Plastids in the trichoblasts gradually become colorless and of less complex structure with increasing distance from the root tip, in contrast to those in adjacent epidermal cells. The trichoblasts do not divide, but they elongate to a considerable extent in the most distal 3000 μ of the root tip and less extensively in the next 3000 μ. By contrast the sister cells and their products divide, but the individual products do not become markedly longer than the mother cell until situated more than 3000 μ from the root tip, when they undergo extensive elongation. The trichoblasts are thus characterized by delayed maturation and inhibition of cytokinesis; it is suggested that delayed maturation is a necessary prerequisite for differentiation of root hairs in this and other species.     

Stage-dependent border cell and carbon flow from roots to rhizosphere

Rising CO(2) levels in the atmosphere have drawn attention to the important role of soil in sequestering carbon. This project goal was to quantify soil carbon deposition associated with border cell release and exudation from root growth zones. Carbon was measured with a Carlo Erba C/N analyzer in soil from the rhizosphere of mature grasses and, in separate experiments, in soil collected around root growth zones. Root border cells in "rhizosphere soil" (silica sand) were counted using a compound microscope after soil sonication and extraction with surfactant. For sand-grown Bromus carinatus, Zea mays, and Cucumis sativus, young seedlings (with roots shorter than 2 cm) released thousands of border cells, while older root tips released only hundreds. For a variety of native annual and perennial grasses and invasive annual grasses (Nassella pulchra, B. carinatus, B. diandrus, B. hordeaceus, Vulpia microstachys, Aegilops triuncialis, Lolium multiflorum, Zea mays), the rhizosphere of mature root systems contained between 18 and 32 μg C g(-1) sand more than that of the unplanted controls. Spatial analysis of the rhizosphere around the cucumber growth zone confirmed C enrichment there. The root tip provided C to the rhizosphere: 4.6 μg C in front of the growing tip, with the largest deposition, 20.4 μg C, to the rhizosphere surrounding the apical 3 mm (root cap/meristem). These numbers from laboratory studies represent the maximum C that might be released during flooding in soils. Scaling up from the organ scale to the field requires a growth analysis to quantify root tip distributions in space and time.     

Vessel-diameter distribution in roots of grapevines (Vitis vinifera L. cv. Shiraz)

The distribution frequency patterns of diameter of xylem vessels and percentage of total predicted axial conductances were studied in 190-day and 212-day-old main roots of grapevine (Vitis vinifera L. cv. Shiraz) grown under well-watered and stressed conditions. The protoxylem were the first to mature and were responsible for most of the theoretical conductance in root segments between the tip and 2.5 cm from the tip. Some large xylem vessels retained cross walls and protoplasm up to 22.5 cm from the tip. Statistical tests using the Kolmogorov-Smirnov two sample test showed that the pattern of distribution frequency of xylem vessels classified in different diameter classes varied with distance from the root tip. The distribution frequency of xylem vessels was similar in both well-watered and stressed plants from the tip up to 15 cm from the tip. At distances further from the tip the distribution frequency of xylem vessels of well-watered plants was significantly different from that of stressed plants, with the former having more larger vessels than the latter. The pattern of vessel distribution frequency was different from that of percent total axial conductance (K_h) predicted with fewer large vessels carrying most of the axial flow.     

Aluminium localization in root tips of the aluminium-accumulating plant species buckwheat (Fagopyrum esculentum Moench)

Aluminium (Al) uptake and transport in the root tip of buckwheat is not yet completely understood. For localization of Al in root tips, fluorescent dyes and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) were compared. The staining of Al with morin is an appropriate means to study qualitatively the radial distribution along the root tip axis of Al which is complexed by oxalate and citrate in buckwheat roots. The results compare well with the distribution of total Al determined by LA-ICP-MS which could be reliably calibrated to compare with Al contents by conventional total Al determination using graphite furnace atomic absorption spectrometry. The Al localization in root cross-sections along the root tip showed that in buckwheat Al is highly mobile in the radial direction. The root apex predominantly accumulated Al in the cortex. The subapical root section showed a homogenous Al distribution across the whole section. In the following root section Al was located particularly in the pericycle and the xylem parenchyma cells. With further increasing distance from the root apex Al could be detected only in individual xylem vessels. The results support the view that the 10 mm apical root tip is the main site of Al uptake into the symplast of the cortex, while the subapical 10-20 mm zone is the main site of xylem loading through the pericycle and xylem parenchyma cells. Progress in the better molecular understanding of Al transport in buckwheat will depend on the consideration of the tissue specificity of Al transport and complexation.