Pectin esterification is spatially regulated both within cell walls and between developing tissues of root apices

Monoclonal antibodies recognizing un-esterified (JIM5) and methyl-esterified (JIM7) epitopes of pectin have been used to locate these epitopes by indirect immunofluorescence and immunogold electron microscopy in the root apex of carrot (Daucus carota L.). Both antibodies labelled the walls of cells in all tissues of the developing root apex. Immunogold labelling observed at the level of the electron microscope indicated differential location of the pectin epitopes within the cell walls. The un-esterified epitope was located to the inner surface of the primary cell walls adjacent to the plasma membrane, in the middle lamella and abundantly to the outer surface at intercellular spaces. In contrast, the epitope containing methyl-esterified pectin was located evenly throughout the cell wall. In root apices of certain other species the JIM5 and JIM7 epitopes were found to be restricted to distinct tissues of the developing roots. In the root apex of oat (Avena sativa L.), JIM5 was most abundantly reactive with cell walls at the region of intercellular spaces of the cortical cells. JIM7 was reactive with cells of the cortex and the stele. Neither epitope occurred in walls of the epidermal or root-cap cells. These pattern of expression were observed to derive from the very earliest stages of the development of these tissues in the oat root meristem and were maintained in the mature root. In the coleoptile and leaf tissues of oat seedlings, JIM5 labelled all cells abundantly whereas JIM7 was unreactive. Other members of the Gramineae and also the Chenopodiaceae are shown to express similar restricted spatial patterns of distribution of these pectin epitopes in root apices.Abbreviations CDTA 1,2-diaminocyclohexane tetraacetic acid - RG rhamnogalacturonan J.P.K. was supported by the Agricultural and Food Research Council Cell Signalling and Recognition Programme. We thank J. Cooke and N. Stacey for technical assistance, H.A. Schols, Drs. P. Albersheim and A. Darvill for pectic polysaccharides, and Dr. R.R. Selvendran and M. McCann for useful discussions.     

Tissue-related changes in methyl-esterification of pectic polysaccharides in cauliflower (Brassica oleracea L. var. botrytis) stems

Pectic substances are a major component of cell walls in vegetable plants and have an important influence on plant food texture. Cauliflower (Brassica oleracea L. var. botrytis) stem sections at different regions of the mature plant stem have been monitored for tissue-related changes in the native pectic polysaccharides. Chemical analysis detected appreciable differences in the degree of methyl-esterification (ME) of pectic polysaccharides. About 65% of galacturonic acid (GalpA) residues were methyl-esterified in floret tissues. Relative ME showed a basipetal decrease, from 94% in the upper stem to 51% in the lower-stem vascular tissues. The decrease was not related to a basipetal increase in glucuronic acid (GlcpA) residues. The monoclonal antibodies, JIM 5 and JIM 7, produced distinct labelling patterns for the relatively low-methyl-esterified and high-methyl-esterified pectin epitopes, respectively. Labelling was related to cell type and tissue location in the stem. Floret cell walls contained epitopes for both JIM 5 and JIM 7 throughout the wall. Stem vascular tissues labelled more strongly with JIM 5. Whereas pith parenchyma in the upper stem labelled more strongly with JIM 7, in the lower-stem pith parenchyma, JIM 5 labelling predominated. Localization of pectic polysaccharide epitopes in cell walls provides an insight into how structural modifications might relate to the textural and nutritional properties of cell walls. Received: 16 August 1997 / Accepted: 20 December 1997     

Localization of cell wall proteins in relation to the developmental anatomy of the carrot root apex

A panel of monoclonal antibodies that recognize a class of cell wall proteins, related to the hydroxyproline-rich glycoproteins, has been assembled and characterized in relation to their restricted patterns of binding amongst the cells comprising the carrot root apex. The occurrence of the epitopes at the surface of cells and intercellular spaces in the region of the apex between the meristematic initials and the region of cell expansion indicates dynamic patterns that reflect aspects of the development of the anatomical pattern. The monoclonal antibody JIM11 reacts with the surface of cells in the central root cap and the region of the meristem. As the cortex/stele boundary becomes established the reactivity is seen in the inner cortical layers and finally in the whole cortex. Later in development the JIM11 epitope is also expressed by two pairs of pericycle cell files adjacent to the phloem region and also by the epidermis. The JIM12 monoclonal antibody is unreactive with cells in the region of the root cap and the meristem but is reactive with intercellular spaces formed at the junction of the oblique and radial walls in the double-layered sectors of the pericycle opposite the xylem poles. This epitope is also transiently expressed by the two phloem sieve tube element mother cells. Later in development JIM12 recognizes the future metaxylem cells. The antibody JIM20 recognizes all the cells and intercellular spaces recognized by JIM11 and JIM12. Immuno-chemical analyses indicate cross-reactivity with carrot taproot extensin and Solanaceous lectins.     

Localization of cell wall polysaccharides in nonarticulated laticifers ofAsclepias speciosa Torr.

Summary Asclepias speciosa Torr, has latex-containing cells known as nonarticulated laticifers. In stem sections of this species, we have analyzed the cell walls of nonarticulated laticifers and surrounding cells with various stains, lectins, and monoclonal antibodies. These analyses revealed that laticifer walls are rich in (1→4) β-D-glucans and pectin polymers. Immunolocalization of pectic epitopes with the antihomogalacturonan antibodies JIM5 and JIM7 produced distinct labeling patterns. JIM7 labeled all cells including laticifers, while JIM5 only labeled mature epidermal cells and xylem elements. Two antibodies, LM5 and LM6, which recognize rhamnogalacturonan I epitopes distinctly labeled laticifer walls. LM6, which binds to a (l→5) α-arabinan epitope, labeled laticifer walls more intensely than walls of other cells. LM5, which recognizes a (1→4) β-D-galac-tan epitope, did not label laticifer segments at the shoot apex but labeled more mature portions of laticifers. Also the LM5 antibody did not label cells at the shoot apical meristem, but as cells grew and matured the LM5 epitope was expressed in all cells. LM2, a monoclonal antibody that binds to β-D-glucuronic acid residues in arabinogalactan proteins, did not label laticifers but specifically labeled sieve tubes. Sieve tubes were also specifically labeled byRicinus communis agglutinin, a lectin that binds to terminal β-D-galactosyl residues. Taken together, the analyses conducted showed that laticifer walls have distinctive cytochemical properties and that these properties change along the length of laticifers. In addition, this study revealed differences in the expression of pectin and arabinogalactan protein epitopes during shoot development or among different cell types.     

Uricase from leaves: its purification and characterization from three different higher plants

Uricase (urate: oxygen oxidoreductase, EC␣1.7.3.3) from leaves of chickpea (Cicer arietimum L.), broad bean (Vicia faba major L.), and wheat (Triticum aestivum L.) has been purified to electrophoretic homogeneity by a procedure which includes xanthine-agarose affinity chromatography as the main step. Purification factors of 74 000–83 000 and recoveries of 80–90% were achieved. Purified preparations had specific activities between 600 and 800 nkat · mg protein⁻¹ (turnover numbers between 4400 and 6400 min⁻¹). The three plant uricases were found by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be tetramers of similar molecular mass (120–130 kDa) and to have identical or similar-sized subunits (32–34 kDa). They also had a similar optimum pH (9–9.5) and showed a hyperbolic kinetics with K _m values from 9–24 μM. All of them showed similar responses to putative activators/inhibitors. Oxonate, xanthine and, to a lesser extent, neocuproin inhibited uricase activity, whereas allantoin, ammonium, citrulline and glutamine did not. The three leaf uricases lacked catalase activity and were not activated by cadaverine. None of the three plant enzymes cross-reacted with anti-uricase monoclonal antibodies from soybean nodules or anti-uricase polyclonal antibodies from Chlamydomonas reinhardtii or rat liver. These results are consistent with the view that uricase in plants is probably a unique enzyme which is expressed at very low level in leaves. Received: 28 October 1996 / Accepted: 8 January 1997     

Root characteristics of representative Mediterranean plant species and their erosion-reducing potential during concentrated runoff

Gully erosion is an important soil degradation process in Mediterranean environments. Revegetation strategies for erosion control rely in most cases on the effects of the above-ground biomass on reducing water erosion rates, whereas the role of the below-ground biomass is often neglected. In a Mediterranean context, the above-ground biomass can temporally disappear because of fire or overgrazing and when concentrated flow erosion occurs, roots can play an important role in controlling soil erosion rates. Unfortunately, information on root characteristics of Mediterranean plants, growing on semi-natural lands, and their effects on the topsoil resistance to concentrated flow erosion is lacking. Therefore, typical Mediterranean grass, herb, reed, shrub and tree root systems of plants growing in habitats that are prone to concentrated flow erosion (i.e. in ephemeral channels, abandoned fields and steep badland slopes) are examined and their erosion-reducing potential was evaluated. Root density (RD), root length density (RLD) and root diameters are measured for 26 typical Mediterranean plant species. RD values and root diameter distribution within the upper 0.10–0.90 m of the soil profile are then transformed into relative soil detachment rates using an empirical relationship in order to predict the erosion-reducing effect of root systems during concentrated runoff. Comparing the erosion-reducing potential of different plant species allows ranking them according to their effectiveness in preventing or reducing soil erosion rates by concentrated flow. RD in the 0.10 m thick topsoil ranges between 0.13 kg m⁻³ for Bromus rubens (L.) and 19.77 kg m⁻³ for Lygeum spartum (L.), whereas RLD ranges between 0.01 km m⁻³ for Nerium oleander (L.) and 120.43 km m⁻³ for Avenula bromoides ((Gouan) H. Scholz.) Relative soil detachment rates, compared to bare soils, range between 0.3 × 10^-12 and 0.7 for the 0.10 m thick topsoil. The results show that grasses such as Helictotrichon filifolium ((Lag.) Henrard), Piptatherum miliaceum ((L.) Coss.), Juncus acutus (L.), Avenula bromoides ((Gouan) H. Scholz), Lygeum spartum (L.) and Brachypodium retusum ((Pers.) Beauv.) have the highest potential to reduce soil erosion rates by concentrated flow in the 0–0.1 m topsoil. But also shrubs such as Anthyllis cytisoides (L.) and Tamarix canariensis (Willd.), having high root densities in the topsoil, can reduce erosion rates drastically. Among the species growing in channels, Juncus acutus (L.) has the highest erosion reducing potential, whereas Phragmites australis (Cav.) is the least effective. On abandoned fields, Avenula bromoides ((Gouan) H. Scholz) and Plantago albicans (L.) are the most effective species in reducing concentrated flow erosion rates, while Thymelaea hirsuta (L. (Endl.)) and Bromus rubens (L.) perform the worst. On steep badland slopes, Helictotrichon filifolium ((Lag.) Henrard) and Anthyllis cytisoides (L.) perform the best in the analysis of erosion reducing potential, while Ononis tridentata (L.) is the least effective species. These findings have implications for ecological restoration and management of erosion-prone slopes.     

Arabinogalactan-proteins in Cichorium somatic embryogenesis: effect of β-glucosyl Yariv reagent and epitope localisation during embryo development

 Direct somatic embryogenesis was induced in root tissues of the Cichorium hybrid `474' (C. intybus L. var. sativum×C. endivia L. var. latifolia). Addition of β-d-glucosyl Yariv reagent (βGlcY), a synthetic phenylglycoside that specifically binds arabinogalactan-proteins (AGPs), to the culture medium blocked somatic embryogenesis in a concentration-dependent manner with complete inhibition of induction occurring at 250 μM βGlcY. The AGP-unreactive α-d-galactosyl Yariv reagent had no biological activity in this system. Upon transfer of 250 μM βGlcY-treated roots to control conditions, somatic embryogenesis was recovered with a time course similar to that of control roots. The βGlcY penetrated roots and bound abundantly to developing somatic embryos, to the root epidermis and the stele. Immunofluorescence and immunogold labelling using monoclonal antibodies (JIM13, JIM16 and LM2) revealed that AGPs were localised in the outer cell walls peripheral cells of the globular embryo. A spatio-temporal expression of AGPs appeared to be associated with differentiation events in the somatic embryo during the transition from the globular stage to the torpedo stage. To verify βGlcY specificity, molecules that bound βGlcY were extracted from treated conditioned medium and identified as AGPs by using the same monoclonal antibodies. In addition, AGPs were found to be abundantly present in the medium during embryogenic culture. All of these results establish the implication of AGPs in embryo development, and their putative role in somatic embryogenesis is discussed. Received: 26 August 1999 / Accepted: 28 January 2000     

Immunohistochemical analysis of cell wall hydroxyproline-rich glycoproteins in the roots of resistant and susceptible wax gourd cultivars in response to Fusarium oxysporum f. sp. Benincasae infection and fusaric acid treatment

Hydroxyproline-rich glycoproteins (HRGPs) play a defensive role in host–pathogen interactions. However, specific roles of individual HRGPs in plant defense against pathogen are poorly understood. Changes in extracellular distribution and abundance of individual cell wall HRGPs were investigated on root sections of two wax gourd (Benincasa hispida Cogn.) cultivars (Fusarium wilt resistant and susceptible, respectively), which were analyzed by immunolabelling with 20 monoclonal antibodies recognizing different epitopes of extensins and arabinogalactan proteins (AGPs) after being inoculated with Fusarium oxysporum f. sp. Benincasae or treated with fusaric acid (FA). These analyses revealed the following: (1) The levels of JIM11 and JIM20 interacting extensins were higher in the resistant cultivar. Either treatment caused a dramatic decrease in signal in both cultivars, but some new signal appeared in the rhizodermis. (2) The AGPs or rhamnogalacturonan containing CCRCM7-epitope were enhanced in the resistant cultivar, but not in the susceptible one by either treatment. (3) Either treatment caused a slight increase in the levels of the AGPs recognized by LM2 and JIM16, but there were no differences between two cultivars. (4) The MAC204 signal nearly disappeared after FA treatment, but this was not the case with pathogen attack. (5) The LM14 signal slightly decreased after both treatments in both cultivars, but a less decrease was observed with the resistant cultivar. These results indicate that the CCRCM7 epitope likely contributed to the resistance of wax gourd to this pathogen, and JIM11 and JIM20 interacting extensins as well as LM2, LM14, MAC204 and JIM16 interacting AGPs were involved in the host–pathogen interaction.     

Cell-specific expression of two arabinogalactan protein epitopes recognized by monoclonal antobodies JIM8 and JIM13 in maize roots

Summary The cell-specific expression of two arabinogalactan protein (AGP) epitopes recognized by monoclonal antibodies JIM8 and JIM13 is reported in maize roots. Employing immunofluorescence and immunogold electron microscopy, the JIM8 antibody was shown to label exclusively protophloem sieve elements, while the JIM13 antibody labelled sieve elements very strongly and adjacent pericycle and companion cells, as well as sloughing root cap cells less strongly. Since the labelling of sieve elements with JIM8 antibody was specific and did not spread to other cell types during root development, it is concluded that this AGP epitope can serve as a specific marker of these specialized cells within the maize root. In the case of the AGP epitope recognized by JIM13 antibody, part of the immunofluorescence label was also found to be associated with cytoplasmic strands in the pericycle and sloughing root cap cells. Immunogold-labelling of sieve elements revealed the association of both AGP epitopes (JIM8 and JIM13) with cortical sieve element reticulum and plasma membranes. Labelling of sieve element reticulum was prominent at its domains of adhesion to the plasma membrane, P-type plastids, and mitochondria. Based on our subcellular studies, we propose a new function of AGP epitopes in endomembrane recognition and adhesion within the sieve elements of maize roots.Abbreviations AGP arabinogalactan protein - SER sieve element reticulum     

Root tensile strength and root distribution of typical Mediterranean plant species and their contribution to soil shear strength

In Mediterranean environments, gully erosion is responsible for large soil losses. It has since long been recognized that slopes under vegetation are much more resistant to soil erosion processes compared to bare soils and improve slope stability. Planting or preserving vegetation in areas vulnerable to erosion is therefore considered to be a very effective soil erosion control measure. Re-vegetation strategies for erosion control rely in most cases on the effects of the above-ground biomass in reducing water erosion rates, whereas the role of the below-ground biomass is often neglected or underestimated. While the above-ground biomass can temporally disappear in semi-arid environments, roots may still be present underground and play an important role in protecting the topsoil from being eroded. In order to evaluate the potential of plant species growing in Mediterranean environments to prevent shallow mass movements on gully or terrace walls, the root reinforcement effect of 25 typical Mediterranean matorral species (i.e. shrubs, grasses herbs, small trees) was assessed, using the simple perpendicular model of Wu et al. (Can Geotech J 16:19–33, 1979). As little information is available on Mediterranean plant root characteristics, root distribution data were collected in SE-Spain and root tensile strength tests were conducted in the laboratory. The power root tensile strength–root diameter relationships depend on plant species. The results show that the shrubs Salsola genistoides Juss. Ex Poir. and Atriplex halimus L. have the strongest roots, followed by the grass Brachypodium retusum (Pers.) Beauv. The shrubs Nerium oleander L. and the grass Avenula bromoides (Gouan) H. Scholz have the weakest roots in tension. Root area ratio for the 0–0.1 m topsoil ranges from 0.08% for the grass Piptatherum miliaceum (L.) Coss to 0.8% for the tree Tamarix canariensis Willd. The rush Juncus acutus L. provides the maximum soil reinforcement to the topsoil by its roots (i.e. 304 kPa). Grasses also increase soil shear strength significantly (up to 244 kPa in the 0–0.1 m topsoil for Brachypodium retusum (Pers.) Beauv.). The shrubs Retama sphaerocarpa (L.) Boiss. and Anthyllis cytisoides L. are increasing soil shear strength to a large extent as well (up to 134 and 160 kPa respectively in the 0–0.10 m topsoil). Whereas grasses and the rush Juncus acutus L. increase soil shear strength in the topsoil (0–0.10 m) to a large extent, the shrubs Anthyllis cytisoides (L.), Retama sphaerocarpa (L.) Boiss., Salsola genistoides Juss. Ex Poir. and Atriplex halimus L. strongly reinforce the soil to a greater depth (0–0.5 m). As other studies reported that Wu’s model overestimates root cohesion values, reported root cohesion values in this study are maximum values. Nevertheless, the calculated cohesion values are used to rank species according to their potential to reinforce the soil.     

Root Strength and Root Area Ratio of Forest Species in Lombardy (Northern Italy)

Forest vegetation is known to increase hillslope stability by reinforcing soil shear resistance and by influencing hydrologic conditions of soil. Although the importance of plant root systems for hillslope stability has received considerable attention in recent years, the quantification of such an effect needs more investigation. In this paper, we present a synthesis of the data gathered in the last 5 years for some species in different locations of the Alps and Prealps of Lombardy (Northern Italy) with the aim to increase our knowledge on root tensile strength and on Root Area Ratio distribution within the soil. Concerning root tensile strength we developed tensile strength–diameter relationships for eight species: green alder (Alnus viridis(Chaix) D.C.), beech (Fagus sylvatica L.), red willow (Salix purpurea L.), goat willow (Salix caprea L.), hazel (Corylus avellana L.), European ash (Fraxinus excelsior L.), Norway spruce (Picea abies (L.) Karst.) and European larch (Larix decidua Mill.). Results show a great variability among the different species and also for the same species. In general, however, root strength (in terms of tension) tends to decrease with diameter according to a power law, as observed by other Authors. Comparing the power law fitting curves for the considered species, it can be observed that they fall in a relatively narrow band, with the exception of hazel, which appears the most resistant. Concerning the evaluation of root distribution within the soil we estimated the Root Area Ratio (the ratio between the area occupied by roots in a unit area of soil) according to its depth for five species (beech, Norway spruce, European larch, mixed hazel and ash) in three locations of Lombardy. Results show that there is a great variability of root density for the same species well as for different points at the same locality. The general behaviour of root density, in any case, is to decrease with depth according to a gamma function for all the studied species. The results presented in this paper contribute to expanding the knowledge on root resistance behaviour and on root density distribution within the soil. The studied location have allowed the implementation of soil–root reinforcement models and the evaluation of the vegetation contribution to soil stability.     

A Comparative Anatomical Study of Organogenesis in Cultured Tissues of Maize, Wheat and Oats

Five cereals and two related grasses were tested for adventitious shoot production from tissue cultures using methods concordant with those reported to be successful for cereals. The five cereals I wheat (Triticum aestivum L.), oats (Avena sativa L.), and maize (Zea mays L.) Pioneer hybrid 3369A, the Bolivian race Pororo and the Equadorian race Chococenõl were all found to proliferate in culture through an aberrant root-like mechanism of growth which had the external appearance of callus. Two related species, teosinte (Zea mexicana Reeves and Mangelsdorf) and tripsacum (Tripsacum dactyloides L.), were less successful in culture, but grew in the same way. Oats, and probably Pororo and Chococeño, initiated presumptive shoot meristems directly from root vascular tissues within this root-like growth. Hybrid maize and wheat initiated no shoot meristems and produced only roots. The occasional shoot production observed in wheat was discounted as simple carryover of existing shoot apices from the primary embryo cultures. This study suggests that the incidence of shoot regeneration in cultures of these cereals may be related more directly to adventitious bud formation on roots than to any controlled de novo organogenesis from undifferentiated callus.     

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     

The monoclonal antibody JIM19 modulates abscisic acid action in barley aleurone protoplasts

A panel of hybridoma products generated against pea (Pisum sativum L.) guard-cell protoplasts has been assayed for anti-abscisic acid (ABA) biological activity in barley (Hordeum vulgare L.) aleurone protoplasts. The effects of the antibodies on ABA-induced accumulation of mRNA transcribed from RAB-16, a gene responsive to ABA, were determined. Most of the antibodies, and culture medium, had no effect, but five monoclonal antibodies (MAbs) were found to inhibit ABA-induced RAB-16 gene expression and one MAb enhanced it. The effects of one inhibitory MAb, JIM19, were studied in some detail. These effects were specific to ABA-induced events, as incubation with JIM19 had no effect on the expression of a constitutively-expressed gene, GAPDH, encoding glyceraldehyde-3-phosphate dehydrogenase, and only a slight effect on the production of -amylase induced by gibberellic acid. Increasing concentrations of ABA in the incubation medium partly overcame the inhibitory effect of JIM19. Immunolabelling and biological activity remained together during immuno-purification of JIM19 from hybridoma culture supernatant. Immunoblotting of JIM19 to membrane preparations from barley aleurone protoplasts revealed that JIM19 recognised a number of proteins.Abbreviations ABA abscisic acid - GA gibberellin - GA₃ gibberellic acid - GAPDH gene encoding glyceraldehyde-3-phosphate dehydrogenase - GCP guard-cell protoplast - MAb monoclonal antibody - RAB (gene) responsive to ABA We thank the Agricultural and Food Research Council and The Nuffield Foundation for financial support, Professor Keith Roberts (John Innes Institute, Norwich, UK) for advice and generous use of his laboratory and Jan Peart (John Innes Institute) for animal cell culture. S.J.N. is grateful to Professor Colin Hawkes (University of the West of England, Bristol) for his continued support of this project.     

Secondary root growth in Rhynchosia edulis Griseb. (Leguminosae): Origin of cambia and their products

This study focuses on the development of a secondary root structure in Rhynchosia edulis Griseb. (Leguminosae). Its principal objectives are (i) to study the origin of cambia and the nature of their products; (ii) to correlate root structure to habitat; and (iii) to compare this anatomy with that of other Leguminosae species growing in the same environment. Serial transverse cuts of the main root show that the secondary root structure in this species results from several phenomena, namely (1) a cambium arising from procambial and pericycle cells; (2) a lateral meristem producing cell layers from the periphery towards the inner part of the root and from which vascular bundles, whose cambia fuse forming a continuous ring, originate; and (3) the formation of “elliptical cambia” in the mid portion of the root giving rise to vascular bundles in reverse orientation. The comparison of secondary root growth in R. edulis with other root structures in Leguminosae species growing in hilly areas shows different structural patterns. Nonetheless, these different patterns have the same objective: to enlarge storage parenchyma tissue enabling survival within an environment having limited water availability.     

Phenylpropanoids in mycorrhizas of the Pinaceae

Tissue-specific accumulation of phenylpropanoids was studied in mycorrhizas of the conifers, silver fir (Abies alba Mill.), Norway spruce [Picea abies (L.) Karst.], white pine (Pinus strobus L.), Scots pine (Pinus silvestris L.), and Douglas fir [Pseudotsuga menziesii (Mirbel) Franco], using high-performance liquid chromatography and histochemical methods. The compounds identified were soluble flavanols (catechin and epicatechin), proanthocyanidins (mainly dimeric catechins and/or epicatechins), stilbene glucosides (astringin and isorhapontin), one dihydroflavonol glucoside (taxifolin 3′-O-glucopyranoside), and a hydroxycinnamate derivative (unknown ferulate conjugate). In addition, a cell wall-bound hydroxycinnamate (ferulate) and a hydroxybenzaldehyde (vanillin) were analysed. Colonisation of the root by the fungal symbiont correlated with the distribution pattern of the above phenylpropanoids in mycorrhizas suggesting that these compounds play an essential role in restricting fungal growth. The levels of flavanols and cell wall-bound ferulate within the cortex were high in the apical part and decreased to the proximal side of the mycorrhizas. In both Douglas fir and silver fir, which allowed separation of inner and outer parts of the cortical tissues, a characteristic transversal distribution of these compounds was found: high levels in the inner non-colonised part of the cortex and low levels in the outer part where the Hartig net is formed. Restriction of fungal growth to the outer cortex may also be achieved by characteristic cell wall thickening of the inner cortex which exhibited flavanolic wall infusions in Douglas fir mycorrhizas. Long and short roots of conifers from natural stands showed similar distribution patterns of phenylpropanoids and cell wall thickening compared to the respective mycorrhizas. These results are discussed with respect to co-evolutionary adaptation of both symbiotic partners regarding root structure (anatomy) and root chemistry. Received: 25 May 1998 / Accepted: 6 November 1998     

Localization of pectins in the pollen tube wall of Ornithogalum virens L. Does the pattern of pectin distribution depend on the growth rate of the pollen tube?

Monoclonal antibodies that recognize pectins were used for the localization of esterified (JIM7) and acidic, unesterified (JIM5) forms of pectin in pollen tube walls of Ornithogalum virens L. (x = n = 3). The results indicated that the distribution of the two forms of pectin in the pollen tube wall depended on the medium (liquid or solid) used for pollen germination. In pollen tubes grown in the liquid medium, the localization of JIM7 was limited to the very tip of the pollen tube, whereas the localization of JIM5 indicated a uniform distribution of unesterified pectins in the very tip of the tube and along the subapical parts of the tube wall. In tubes germinated on the medium stabilized with agar (1–2%) the localization of JIM7 and JIM5 indicated the presence of both forms of pectin in the tube tip and along the whole length of the pollen tube wall in a ring-like pattern. Thus, the localization of esterified pectins in the sub-apical part of the pollen tube wall, below the apex of the tube, is described for the first time. Measurements of the growth rates of pollen tubes growing on the two types of medium indicated that oscillations in tube growth rate occur but these do not coincide with the pattern of pectin distribution in the tube wall. Our results complement the previous data obtained for the localization of JIM5 and JIM7 in pollen tube walls of other plant species. (Y.-Q. Li et al. 1994, Sex Plant Reprod 7: 145–150) and provide new insight into an understanding of the construction of the pollen tube wall and the physiology of pollen grain germination. Received: 25 January 1999 / Accepted: 23 June 1999     

Chromium accumulation, translocation and chemical speciation in vegetable crops

Trivalent chromium (Cr³⁺) is essential for animal and human health, whereas hexavalent Cr (CrO₄ ²⁻) is a potent carcinogen and extremely toxic to animals and humans. Thus, the accumulated Cr in food plants may represent potential health hazards to animals and humans if the element is accumulated in the hexavalent form or in high concentrations. This study was conducted to determine the extent to which various vegetable crops absorb and accumulate Cr³⁺ and CrO₄ ²⁻ into roots and shoots and to ascertain the different chemical forms of Cr in these tissues. Two greenhouse hydroponic experiments were performed using a recirculating-nutrient culture technique that allowed all plants to be equally supplied with Cr at all times. In the first experiment, 1 mg L⁻¹ Cr was supplied to 11 vegetable plant species as Cr³⁺ or CrO₄ ²⁻, and the accumulation of Cr in roots and shoots was compared. The crops tested included cabbage (Brassica oleracea L. var. capitata L.), cauliflower (Brassica oleracea L. var. botrytis L.), celery (Apium graveolens L. var. dulce (Mill.) Pers.), chive (Allium schoenoprasum L.), collard (Brassica oleracea L. var. acephala DC.), garden pea (Pisum sativum L.), kale (Brassica oleracea L. var. acephala DC.), lettuce (Lactuca sativa L.), onion (Allium cepa L.), spinach (Spinacia oleracea L.), and strawberry (Fragaria ×  ananassaDuch.). In the second experiment, X-ray absorption spectroscopy (XAS) analysis on Cr in plant tissues was performed in roots and shoots of various vegetable plants treated with CrO₄ ²⁻ at either 2 mg Cr L⁻¹ for 7 d or 10 mg Cr L⁻¹ for 2, 4 or 7 d. The crops used in this experiment included beet (Beta vulgaris L. var. crassa (Alef.) J. Helm), broccoli (Brassica oleracea L. var. Italica Plenck), cantaloupe (Cucumis melo L. gp. Cantalupensis), cucumber (Cucumis sativus L.), lettuce, radish (Raphanus sativus L.), spinach, tomato (Lycopersicon lycopersicum (L.) Karsten), and turnip (Brassica rapa L. var. rapifera Bailey). The XAS speciation analysis indicates that CrO₄ ²⁻ is converted in the root to Cr³⁺ by all plants tested. Translocation of both Cr forms from roots to shoots was extremely limited and accumulation of Cr by roots was 100-fold higher than that by shoots, regardless of the Cr species supplied. Highest Cr concentrations were detected in members of the Brassicaceae family such as cauliflower, kale, and cabbage. Based on our observations and previous findings by other researchers, a hypothesis for the differential accumulation and identical translocation patterns of the two Cr ions is proposed. Received: 27 February 1998 / Accepted: 2 April 1998     

An AGP epitope distinguishes a central metaxylem initial from other vascular initials in theArabidopsis root

Summary The developmental expression of an arabinogalactan protein (AGP) recognised by a monoclonal antibody, JIM 13, is described inArabidopsis roots. It is expressed in the single initial of the central metaxylem vessel that lies immediately above the four central cells of the quiescent centre. AGP expression spreads in a non-clona] fashion to neighbouring cell files as they mature. The AGP first appears in other pre-metaxylem elements and subsequently in the layer of parenchyma cells on either side of the xylem axis. Later, it spreads to the entire ring of eight endodermal cell files and to those pericycle cell files next to the xylem pole. It is postulated that this epitope reflects an underlying set of cell signalling events that may be involved in defining local positional values of cells important in establishing cellular patterns in the root.     

Nectar-carbohydrate production and composition vary in relation to nectary anatomy and location within individual flowers of several species of Brassicaceae

Nectar-carbohydrate production and composition were investigated by high-performance liquid chromatography and enzymology in nine species from five tribes of the Brassicaceae. In six species (Arabidopsis thaliana (L.) Heynh., Brassica napus L., B. rapa L., Lobularia maritima (L.) Desv., Raphanus sativus L., Sinapis arvensis L.) that produced nectar from both lateral nectaries (associated with the short stamens) and median nectaries (outside the long stamens), on average 95% of the total nectar carbohydrate was collected from the lateral ones. Nectar from these glands possessed a higher glucose/fructose ratio (usually 1.0–1.2) than that from the median nectaries (0.2–0.9) within the same flower. Comparatively little sucrose was detected in any nectar samples except from Matthiola bicornus (Sibth. et Sm.) DC., which possessed lateral nectaries only and produced a sucrose-dominant exudate. The anatomy of the nectarial tissue in nectar-secreting flowers of six species, Hesperis matronalis L., L. maritima, M. bicornus, R. sativus, S. arvensis, and Sisymbrium loeselii L., was studied by light and scanning-electron microscopy. Phloem alone supplied the nectaries. However, in accordance with their overall nectar-carbohydrate production, the lateral glands received relatively rich quantities of phloem that penetrated far into the glandular tissue, whereas median glands were supplied with phloem that often barely innervated them. All nectarial tissue possessed modified stomata (with the exception of the median glands of S. loeselii, which did not produce nectar); further evidence was gathered to indicate that these structures do not regulate nectar flow by guard-cell movements. The numbers of modified stomata per gland showed no relation to nectar-carbohydrate production. Taken together, the data on nectar biochemistry and nectary anatomy indicate the existence of two distinct nectary types in those Brassicacean species that possess both lateral and median nectaries, regardless of whether nectarial tissue is united around the entire receptacle or not. It is proposed that the term “nectarium” be used to represent collectively the multiple nectaries that can be found in individual flowers. Received: 21 July 1997 / Accepted: 19 September 1997