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PubMed2006

Conditions leading to high CO2 (>5 kPa) in waterlogged-flooded soils and possible effects on root growth and metabolism.

AIMS: Soil waterlogging impedes gas exchange with the atmosphere, resulting in low P(O2) and often high P(CO2). Conditions conducive to development of high P(CO2) (5-70 kPa) during soil waterlogging and flooding are discussed. The scant information on responses of roots to high P(CO2) in terms of growth and metabolism is reviewed. SCOPE: P(CO2) at 15-70 kPa has been reported for flooded paddy-field soils; however, even 15 kPa P(CO2) may not always be reached, e.g. when soil pH is above 7. Increases of P(CO2) in soils following waterlogging will develop much more slowly than decreases in P(O2); in soil from rice paddies in pots without plants, maxima in P(CO2) were reached after 2-3 weeks. There are no reliable data on P(CO2) in roots when in waterlogged or flooded soils. In rhizomes and internodes, P(CO2) sometimes reached 10 kPa, inferring even higher partial pressures in the roots, as a CO2 diffusion gradient will exist from the roots to the rhizomes and shoots. Preliminary modelling predicts that when P(CO2) is higher in a soil than in roots, P(CO2) in the roots would remain well below the P(CO2) in the soil, particularly when there is ventilation via a well-developed gas-space continuum from the roots to the atmosphere. The few available results on the effects of P(CO2) at > 5 kPa on growth have nearly all involved sudden increases to 10-100 kPa P(CO2); consequently, the results cannot be extrapolated with certainty to the much more gradual increases of P(CO2) in waterlogged soils. Nevertheless, rice in an anaerobic nutrient solution was tolerant to 50 kPa CO2 being suddenly imposed. By contrast, P(CO2) at 25 kPa retarded germination of some maize genotypes by 50%. With regard to metabolism, assuming that the usual pH of the cytoplasm of 7.5 was maintained, every increase of 10 kPa CO2 would result in an increase of 75-90 mM HCO3(-) in the cytoplasm. pH maintenance would depend on the biochemical and biophysical pH stats (i.e. regulatory systems). Furthermore, there are indications that metabolism is adversely affected when HCO3(-) in the cytoplasm rises above 50 mM, or even lower; succinic dehydrogenase and cytochrome oxidase are inhibited by HCO3(-) as low as 10 mM. Such effects could be mitigated by a decrease in the set point for the pH of the cytoplasm, thus lowering levels of HCO3(-) at the prevailing P(CO2) in the roots. CONCLUSIONS: Measurements are needed on P(CO2) in a range of soil types and in roots of diverse species, during waterlogging and flooding. Species well adapted to high P(CO2) in the root zone, such as rice and other wetland plants, thrive even when P(CO2) is well over 10 kPa; mechanisms of adaptation, or acclimatization, by these species need exploration.

Acclimatization↗
PubMed2006

Adventitious root production and plastic resource allocation to biomass determine burial tolerance in woody plants from central Canadian coastal dunes.

BACKGROUND AND AIMS: Burial is a recurrent stress imposed upon plants of coastal dunes. Woody plants are buried on open coastal dunes and in forested areas behind active blowouts; however, little is known about the burial responses and adaptive traits of these species. The objectives of this study were: (a) to determine the growth and morphological responses to burial in sand of seven woody plant species native to central Canadian coastal dunes; and (b) to identify traits that determine burial tolerance in these species. METHODS: Field experiments were conducted to determine the responses of each species to burial. Saplings were exposed to burial treatments of 0, 10, 25, 50 and 75 % of their height. Burial responses were evaluated based on regressions of total biomass, height, adventitious root production and percentage allocation to shoot, root and adventitious root biomass on percentage burial. KEY RESULTS: Pinus strobus and Picea glauca lacked burial tolerance. In response to the burial gradient, these species showed a strong linear decline in total biomass, minimal adventitious root production that peaked at moderate levels (25-50 % burial) and no change in allocation to shoots vs. roots. The tolerant species Juniperus virginiana, Thuja occidentalis and Picea mariana showed a quadratic response to burial, with little change in biomass up to 50 % burial, but a large decline at 75 %. These species produced abundant adventitious roots up to 50 % burial, but did not alter allocation patterns over the range of burial levels. Populus balsamifera and Salix cordata were stimulated by burial. These species showed linear increases in biomass with increasing burial, produced copious adventitious roots across the gradient and showed a clear shift in allocation to vertical shoot growth and adventitious root production at the expense of the original roots under high burial conditions. CONCLUSIONS: Adventitious root production and plastic resource allocation to biomass are adaptive traits of coastal dune woody plants in central Canada, and provide a basis for assessing burial tolerance in woody plants on coastal dunes throughout the world.

Adaptation, Physiological↗
PubMed2004

Root-crown ratios of permanent teeth in a healthy Finnish population assessed from panoramic radiographs.

An unfavourable root-crown (R/C) ratio caused by short dental roots may result from a developmental deficiency, root resorption after orthodontic treatment, or dental trauma. In the assessment of root shortening, subjective grading has often been used. For objective tooth measurements, varying materials and methods may make the results impossible to compare. This study used a simple, objective method to assess the R/C ratio (relative root length) of mature permanent teeth from panoramic radiographs (PRGs), tested its reproducibility and calculated the mean values of R/C ratios and their variations in a healthy Caucasian (Finnish) population. Two thousand seven hundred and seventy-nine teeth were measured on 108 PRGs. The intra- and inter-examiner reproducibility of the assessment method was good (Pearson correlation coefficients 0.87 and 0.83, respectively; P < 0.001) and the mean R/C ratios did not differ between the repeated measurements (P > 0.05). The biological variance in all cases exceeded the error variance for each tooth. These facts suggest that the method reported in this study can be used in the assessment of the relative root length of 'normal' teeth and its alterations in teeth with developmental or acquired aberrations of dental roots. Males, overall, tended to have higher R/C ratios than females; P-values varied from non-significant to less than 0.01. With the exception of the permanent lateral incisors in males and the permanent second molars in both genders, the ratios of the antagonist teeth were significantly greater in the mandible than in the maxilla (P < 0.05 for the lateral incisors of females; P < 0.001 for all other teeth). Consequently, in quantifying root shortening in developmentally short-rooted teeth, tooth- and gender-specific reference values should be employed. The Finnish R/C data reported here for all teeth except third molars could be used for comparison with other populations, patient groups or individuals where crown-root aberrations are suspected.

Adolescent↗
PubMed2000

Radial hydraulic conductivity along developing onion roots.

Although most studies have shown that water uptake varies along the length of a developing root, there is no consistent correlation of this pattern with root anatomy. In the present study, water movement into three zones of onion roots was measured by a series of mini-potometers. Uptake was least in the youngest zone (mean hydraulic conductivity, Lpr = 1.5 x 10(-7) +/- 0.34 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots) in which the endodermis had developed only Casparian bands and the exodermis was immature. Uptake was significantly greater in the middle zone (Lpr = 2.4 x 10(-7) +/- 0.43 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots) which had a mature exodermis with both Casparian bands and suberin lamellae, and continued at this level in the oldest zone in which the endodermis had also developed suberin lamellae (Lpr = 2.8 x 10(-7) +/- 0.30 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots). Measurements of the hydraulic conductivities of individual cells (Lp) in the outer cortex using a cell pressure probe indicated that this parameter was uniform in all three zones tested (Lp = 1.3 x 10(-6) +/- 0.01 x 10(-6) m MPa-1 s-1; +/- SE, n = 60 cells). Lp of the youngest zone was lowered by mercuric chloride treatment, indicating the involvement of mercury-sensitive water channels (aquaporins). Water flow in the older two root zones measured by mini-potometers was also inhibited by mercuric chloride, despite the demonstrated impermeability of their exodermal layers to this substance. Thus, water channels in the epidermis and/or exodermis of the older regions were especially significant for water flow. The results of this and previous studies are discussed in terms of two models. The first, which describes maize root with an immature exodermis, is the 'uniform resistance model' where hydraulic resistances are evenly distributed across the root cylinder. The second, which describes the onion root with a mature exodermis, is the 'non-uniform resistance model' where resistances can be variable and are concentrated in a certain layer(s) on the radial path.

Aquaporins↗
PubMed2001

Gene expression profiling of two related maize inbred lines with contrasting root-lodging traits.

To assist breeding for increased resistance to root lodging in maize, an attempt was made to identify genes that are associated with root lodging by profiling mRNA expression from two inbreds with contrasting root-related traits. These two inbreds were derived from a common F(2) pool, selfed for several generations and showed 75% relatedness based on 106 genetic markers. Under field conditions, the two inbreds exhibited significant differences in root morphology and resistance to root lodging. Whole root tissue was collected at two developmental stages from inbred 100 and 101 grown in 2 years. RNA was isolated from both the V8 and V12 stages, a few weeks prior to flowering. The RNA samples from the 1997 growing season were analysed by GeneCalling analysis, an open-ended mRNA profiling method. From over 13 500 cDNA fragments detected from each of the V8- and V12-stage samples, 229 and 325 cDNA fragments, respectively, showed greater than 2-fold differences between the two inbred lines. A total of 69 cDNA fragments that showed 2-fold or greater differences for both inbred lines were observed at both developmental stages. The gene identity and expression differences of several cDNA fragments were determined and confirmed by RNA gel blot analysis. Two genes out of five identified were homologous to a cytochrome P450 and the impedance-induced protein, both showing high levels of expression in the roots of lodging resistant lines and low levels in the sensitive lines. These data provide the first clues of genes expressed in the roots during the formative stages of root development associated with root-lodging resistance.

Amino Acid Sequence↗
PubMed2003

Al-induced efflux of organic acid anions is poorly associated with internal organic acid metabolism in triticale roots.

The secretion of organic acid anions from roots has been identified as a mechanism of resistance to Al. However, the process leading to the secretion of organic acid anions is poorly understood. The effect of Al on organic acid metabolism was investigated in two lines of triticale (xTriticosecale Wittmark) differing in Al-induced secretion of malate and citrate and in Al resistance. The site of Al-induced secretion of citrate and malate from a resistant line was localized to the root apices (terminal 5 mm). The levels of citrate (root apices and mature root segments) and malate (mature segments only) in roots increased during exposure to Al, but similar changes were observed in both triticale genotypes. The in vitro activities of four enzymes involved in malate and citrate metabolism (citrate synthase, phosphoenolpyruvate carboxylase, malate dehydrogenase, and NADP-isocitrate dehydrogenase) were similar for sensitive and resistant lines in both root apices and mature root segments. The response of these enzymes to pH did not differ between tolerant and sensitive lines or in the presence and absence of Al. Moreover, cytoplasmic and vacuolar pH were not affected by exposure to Al in either line. Together, these results indicate that the Al-dependent efflux of organic acid anions from the roots of triticale is not regulated by their internal levels in the roots or by the capacity of the root cells to synthesize malate and citrate.

Acids, Acyclic↗
PubMed2004

Water permeability and reflection coefficient of the outer part of young rice roots are differently affected by closure of water channels (aquaporins) or blockage of apoplastic pores.

The relative contribution of the apoplastic and cell-to-cell paths to the overall hydraulic conductivity of the outer part of rice roots (LpOPR) was estimated using a pressure perfusion technique for 30-d-old rice plants (lowland cultivar, IR64, and upland cultivar, Azucena). The technique was based on the perfusion of aerenchyma of root segments from two different zones (20-50 mm and 50-100 mm from the root apex) with aerated nutrient solution using precise pump rates. The outer part of roots (OPR) comprised an outermost rhizodermis, an exodermis, sclerenchyma fibre cells, and the innermost unmodified cortical cell layer. No root anatomical differences were observed for the two cultivars used. Development of apoplastic barriers such as Casparian bands and suberin lamellae in the exodermis were highly variable. On average, matured apoplastic barriers were observed at around 50-70 mm from the root apex. Lignification of the exodermis was completed earlier than that of sclerenchyma cells. Radial water flow across the OPR was impeded either by partially blocking off the porous apoplast with China ink particles (diameter 50 nm) or by closing water channels (aquaporins) in cell membranes with 50 micro M HgCl2. The reduction of LpOPR was relatively larger in the presence of an apoplastic blockage with ink ( approximately 30%) than in the presence of the water channel blocker ( approximately 10%) suggesting a relatively larger apoplastic water flow. The reflection coefficient of the OPR (sigmasOPR) for mannitol significantly increased during both treatments. It was larger when pores of the apoplast were closed, but absolute values were low (overall range of sigmasOPR=0.1-0.4), which also suggested a large contribution of the non-selective, apoplastic path to overall water flow. The strongest evidence in favour of a predominantly apoplastic water transport came from the comparison between diffusional (PdOPR, measured with heavy water, HDO) and osmotic water permeability (PfOPR) or hydraulic conductivity (LpOPR). PfOPR was larger by a factor of 600-1400 compared with P(dOPR). The development of OPR along roots resulted in a decrease of PdOPR by a factor of three (segments taken at 20-50 and 50-100 mm from root apex, respectively). Heat-killing of living cells resulted in an increase of PdOPR for both immature (20-50 mm) and mature (50-100 mm) root segments by a factor of two. Even though both pathways (apoplast and cell-to-cell) contributed to the overall water flow, the findings indicate predominantly apoplastic water flow across the OPR, even in the presence of apoplastic barriers. Low diffusional water permeabilities may suggest a low rate of oxygen diffusion across the OPR from aerenchyma to the outer anaerobic soil medium (low PO2OPR). To date, there are no data on PO2OPR. Provisional data of radial oxygen losses (ROL) across the OPR suggest that, unlike water, rice roots efficiently retain oxygen within the aerenchyma. This ability strongly increases as roots/OPR develop.

Aquaporins↗
PubMed2004

Developmental anatomy and auxin response of lateral root formation in Ceratopteris richardii.

The homosporous fern Ceratopteris richardii exhibits a homorhizic root system where roots originate from the shoot system. These shoot-borne roots form lateral roots (LRs) that arise from the endodermis adjacent to the xylem poles, which is in contrast to flowering plants where LR formation arises from cell division in the pericycle. A detailed study of the fifth shoot-borne root showed that one lateral root mother cell (LRMC) develops in each two out of three successive merophytes. As a result, LRs emerge alternately in two ranks from opposite positions on a parent root. From LRMC initiation to LR emergence, three developmental stages were identified based on anatomical criteria. The addition of auxins (either indole-3-acetic acid or indole-3-butyric acid) to the growth media did not induce additional LR formation, but exogenous applications of both auxins inhibited parent root growth rate. Application of the polar auxin-transport inhibitor N-(1-naphthyl)phthalamic acid (NPA) also inhibited parent root growth without changing the LR initiation pattern. The results suggest that LR formation does not depend on root growth rate per se. The result that exogenous auxins do not promote LR formation in C. richardii is similar to reports for certain species of flowering plants, in which there is an acropetal LR population and the formation of the LRs is insensitive to the application of auxins. It also may indicate that different mechanisms control LR development in non-seed vascular plants compared with angiosperms, taking into consideration the long and independent evolutionary history of the two groups.

Flowers↗
PubMed2006

Root meristems in Medicago truncatula tissue culture arise from vascular-derived procambial-like cells in a process regulated by ethylene.

Leaf explants of Medicago truncatula were used to investigate the origins of auxin-induced root formation. On the application of auxin there is some callus formation (not the massive amount that occurs in response to auxin plus cytokinin) and roots appear shortly after the first visible callus. Histological examination reveals morphologically distinctive sheets of callus cells that emanate from the veins of the leaf explants and, within this cell type, root primordia are produced as well as some vascular tissue cells. What is suggested is that the vein-derived cells (VDCs) are procambial-like and function as pluripotent stem cells with a propensity to form root meristems or vascular tissues in response to added auxin. The development of root primordia from these pluripotent cells was clearly up-regulated by the use of the sickle (skl) mutant, which is a mutant impaired in ethylene signal transduction while the wild type and the sunn mutant, defective in auxin polar transport, produced similar numbers of roots. The skl mutant in generating many more roots concomitantly formed fewer vascular tissues. The root meristems differentiate similarly to normal roots producing a central cylinder of vascular tissue, which connects with the leaf explant veins. The VDCs appear to be derived from the cells of or near the phloem. The leaf observations suggest that a pool of stem cells exist in vascular tissue that, in combination with auxin and perhaps other factors, drive a diversity of plant development outcomes that is species specific. The way auxin interacts with other hormones is a key factor in determining the stem cell fate. The histological data in this study also assist in the interpretation of the molecular analysis of auxin-induced root formation in cultured leaves of M. truncatula.

Ethylenes↗
PubMed2000

Vascular tissue-specific gene expression of xylem sap glycine-rich proteins in root and their localization in the walls of metaxylem vessels in cucumber.

Root-specific cDNAs of glycine-rich protein (cucumber root glycine rich protein-1 and -2; CRGRP-1 and CRGRP-2) were cloned previously by use of an antiserum raised against whole xylem sap of Cucumis sativus. The accumulation of the corresponding mRNA at high levels was detected in the root-hair zone of cucumber tap root [Sakuta et al. (1998) Plant Cell Physiol. 39: 1330]. The RNA gel blot analysis with the CRGRP-1- and -2-specific probes revealed that the CRGRP genes expressed only in root but not at all in aboveground organs. When the localization of these mRNAs were examined by in situ hybridization, CRGRP mRNAs were found only in the parenchyma cells in the central cylinder of young lateral roots and it was most abundant in the cells that surrounded xylem vessels in the root-hair zone of the tap root. In immunoblotting of xylem sap collected from cucumber stem with an antiserum raised against CRGRP-1 that had been produced in an E. coli expression system, the antibodies, which did not cross-react with GRP1.8 of kidney bean, reacted with two proteins, whose mobilities corresponded to those of proteins deduced from the CRGRP-1 and -2 cDNAs. Immunohistochemical staining revealed that the CRGRPs accumulated specifically in the lignified walls of metaxylem vessels in the root, stem and leaf and in the lignified cell walls of perivascular fibers in cucumber stems. Immunostaining was also detected in the walls of metaxylem vessels and in the cell walls of adjacent sclerenchyma in the hypocotyl of kidney bean. These data clearly indicate that the novel glycine-rich proteins were produced in the vascular tissue of the root, transported systemically over a long distance via the xylem sap and immobilized in the walls of metaxylem vessels and sclerechyma cells in aboveground organs.

Cucumis sativus↗
PubMed2002

Measurement carbon dioxide concentration does not affect root respiration of nine tree species in the field.

Inhibition of respiration has been reported as a short-term response of tree roots to elevated measurement CO2 concentration ([CO2]), calling into question the validity of root respiration rates determined at CO2 concentrations that differ from the soil [CO2] in the rooting zone. Our objectives were to validate previous observations of a direct CO2 effect on root respiration in sugar maple (Acer saccharum Marsh.) and to determine if high [CO2] also inhibited root respiration in other tree species. Root respiration rates for nine common North American tree species were measured in the field at ambient soil temperature at both 350 and 1000 microl CO2 l-1. No evidence of direct inhibition of root respiration by elevated measurement [CO2] was found for any of the species tested. The ratio of respiration rates at 1000 and 350 microl CO2 l-1 ranged from 0.97 to 1.07, and the 95% confidence intervals for this ratio included unity for all species tested. Tests of a respiration cuvette used in earlier experiments suggested that gas leakage from the cuvette/IRGA system created an apparent direct CO2 effect on respiration of sugar maple roots when none actually existed. Small sample masses used in those experiments exacerbated the error. Careful attention to the possibility of gas leaks and the avoidance of small sample masses should produce data that will allow researchers to accurately assess whether direct effects of measurement [CO2] exist. Our findings of no direct CO2 effect on respiration of roots of a wide variety of species suggest that such effects may be less common than previously thought for tree roots.

Carbon Dioxide↗
PubMed2002

Ectomycorrhizal fungi and exogenous auxins influence root and mycorrhiza formation of Scots pine hypocotyl cuttings in vitro.

We studied the ability of the ectomycorrhizal (ECM) fungi, Pisolithus tinctorius (Pers.) Coker and Couch and Paxillus involutus (Batsch) Fr. (Strain H), to produce indole-3-acetic acid (IAA) and to affect the formation and growth of roots on Scots pine (Pinus sylvestris L.) hypocotyl cuttings in vitro. Effects of indole-3-butyric acid (IBA) and the auxin transport inhibitor, 2,3,5-triiodobenzoic acid (TIBA), on rooting and the cutting-fungus interaction were also studied. Both fungi produced IAA in the absence of exogenous tryptophan, but the mycelium and culture filtrate of Pisolithus tinctorius contained higher concentrations of free and conjugated IAA than the mycelium and culture filtrate of Paxillus involutus. Inoculation with either fungus or short-term application of culture filtrate of either fungus to the base of hypocotyl cuttings enhanced root formation. Inoculation with either fungus was even more effective in enhancing root formation than treatment of the hypocotyl bases with IBA. Fungal IAA production was not directly correlated with root formation, because rooting was enhanced more by Paxillus involutus than by Pisolithus tinctorius. This suggests that, in addition to IAA, other fungal components play an important role in root formation. Treatment with 5 microM TIBA increased the rooting percentage of non-inoculated cuttings, as well as of cuttings inoculated with Pisolithus tinctorius, perhaps as a result of accumulation of IAA at the cutting base. However, the marked reduction in growth of Pisolithus tinctorius in the presence of TIBA suggests that the effects of TIBA on rooting are complicated and not solely related to IAA metabolism. The high IAA-producer, Pisolithus tinctorius, formed mycorrhizas, and the IBA treatment increased mycorrhizal frequency in this species, whereas TIBA decreased it. Paxillus involutus did not form mycorrhizas, indicating that a low concentration of IAA together with other fungal components were sufficient to stimulate formation and growth of the roots, but not the formation of ECM symbiosis.

Basidiomycota↗
PubMed2003

Developmental stages during the rooting of in-vitro-cultured Quercus robur shoots from material of juvenile and mature origin.

In-vitro-cultured shoots of clones initiated from shoots of the basal parts (BS) and the crown (C) of two mature Quercus robur L. trees were subjected to rooting experiments to relate rooting with shoot topophysical origin. The BS shoots exhibited morphologically juvenile characteristics and rooted more easily after indole-3-butyric acid (IBA) treatment than C shoots. When naphthylphthalamic acid (NPA) was applied to BS shoots, rooting capacity decreased and root emergence was delayed at least 2 days compared with shoots treated with IBA only. During the first days of the rooting process, endogenous indole-3-acetic acid (IAA) concentration was higher in C shoots than in BS shoots, regardless of whether the shoots were treated with NPA. Mitotic figures were observed in cells from the basal part of both BS and C shoots 24 h after IBA treatment. After 4 days of IBA treatment, the occurrence of histological events differed between BS shoots and C shoots. Cells of BS shoots became meristematic, giving rise to meristemoids and root primordia, whereas no differentiation of root meristemoids occurred in cells of C shoots. Thus, although adult oak material (C shoots) is capable of responding to the initial stimulus of auxin during the adventitious rooting process, the endogenous IAA concentration is not the factor limiting rooting in adult material.

Indoleacetic Acids↗
PubMed2003

Hydraulic redistribution by deep roots of a Chihuahuan Desert phreatophyte.

Downward redistribution of soil water through plant roots has important consequences for water and nutrient balance of arid and semi-arid ecosystems. Nevertheless, information on the seasonal patterns and magnitudes of redistribution is lacking for all but a few plant species. We measured sap flow in the taproot and three main lateral roots of a 10-year-old Juglans major Torr. tree, on an ephemeral catchment in southeastern Arizona, to determine how patterns of redistribution respond to pulses of summer precipitation. Groundwater was beyond rooting depth and a hardpan prevented recharge of surface water to deep soil layers. Reverse flow (hydraulic descent) commenced in the taproot and deep lateral roots in early August after a series of moderate precipitation events, and abruptly ceased after all shallow roots were experimentally severed in mid-August. On some days, hydraulic descent continued in the deep lateral roots during periods of daytime transpiration, and the daily volume of hydraulic descent (deep lateral roots plus taproot) ranged from 10 to nearly 60% of daily transpiration. The persistent pattern of reverse flow demonstrates that, in some plants, water potential gradients from soil to leaf during transpiration are often smaller than those between soil layers within the rooting zone. Hydraulic descent may be an important component of the water balance of phreatophytic trees by facilitating root growth in deep soil layers and by transferring water away from shallow-rooted competitors.

Arizona↗
PubMed2006

Electrical measurement of the absorption surfaces of tree roots by the earth impedance method: 1. Theory.

Analysis of plant root systems is difficult under field conditions, especially root systems of adult trees, which are large and complex and include fine absorbing roots as well as conducting coarse roots. Although coarse roots can be visualized by several methods, there are technical difficulties with root quantification. The method presented here focuses on the quantification of absorbing root surfaces through an electrical (the modified earth impedance) method. It is based on the experimentally verified fact that an applied electric current flows from the roots to the soil (or vice versa) through the same interfacial areas and predominantly in the same way as water (water solution of minerals or nutrients) flows from the soil to the tree. Based on the different conductivities of tree tissues and soil, the interfacial area, which represents the absorbing root surfaces (or root absorption zones), can be calculated. Only the theoretical description of the method is presented in this paper: the experimental verification of the method under field conditions is presented in the accompanying paper.

Absorption↗
PubMed2000

Heterologous rhizobial lipochitin oligosaccharides and chitin oligomers induce cortical cell divisions in red clover roots, transformed with the pea lectin gene.

Division of cortical cells in roots of leguminous plants is triggered by lipochitin oligosaccharides (LCOs) secreted by the rhizobial microsymbiont. Previously, we have shown that presence of pea lectin in transgenic white clover hairy roots renders these roots susceptible to induction of root nodule formation by pea-specific rhizobia (C. L. Díaz, L. S. Melchers, P. J. J. Hooykaas, B. J. J. Lugtenberg, and J. W. Kijne, Nature 338:579-581, 1989). Here, we report that pea lectin-transformed red clover hairy roots form nodule primordium-like structures after inoculation with pea-, alfalfa-, and Lotus-specific rhizobia, which normally do not nodulate red clover. External application of a broad range of purified LCOs showed all of them to be active in induction of cortical cell divisions and cell expansion in a radial direction, resulting in formation of structures that resemble nodule primordia induced by clover-specific rhizobia. This activity was obvious in about 50% of the red clover plants carrying hairy roots transformed with the pea lectin gene. Also, chitopentaose, chitotetraose, chitotriose, and chitobiose were able to induce cortical cell divisions and cell expansion in a radial direction in transgenic roots, but not in control roots. Sugar-binding activity of pea lectin was essential for its effect. These results show that transformation of red clover roots with pea lectin results in a broadened response of legume root cortical cells to externally applied potentially mitogenic oligochitin signals.

Cell Division↗
PubMed2001

Agrobacterium rhizogenes-transformed roots of Medicago truncatula for the study of nitrogen-fixing and endomycorrhizal symbiotic associations.

Medicago truncatula, a diploid autogamous legume, is currently being developed as a model plant for the study of root endosymbiotic associations, including nodulation and mycorrhizal colonization. An important requirement for such a plant is the possibility of rapidly introducing and analyzing chimeric gene constructs in root tissues. For this reason, we developed and optimized a convenient protocol for Agrobacterium rhizogenes-mediated transformation of M. truncatula. This unusual protocol, which involves the inoculation of sectioned seedling radicles, results in rapid and efficient hairy root organogenesis and the subsequent development of vigorous "composite plants." In addition, we found that kanamycin can be used to select for the cotransformation of hairy roots directly with gene constructs of interest. M. truncatula composite plant hairy roots have a similar morphology to normal roots and can be nodulated successfully by their nitrogen-fixing symbiotic partner, Sinorhizobium meliloti. Furthermore, spatiotemporal expression of the Nod factor-responsive reporter pMtENOD11-gusA in hairy root epidermal tissues is indistinguishable from that observed in Agrobacterium tumefaciens-transformed lines. M. truncatula hairy root explants can be propagated in vitro, and we demonstrate that these clonal lines can be colonized by endomycorrhizal fungi such as Glomus intraradices with the formation of arbuscules within cortical cells. Our results suggest that M. truncatula hairy roots represent a particularly attractive system with which to study endosymbiotic associations in transgenically modified roots.

Fabaceae↗
PubMed2002

Oomycete plant pathogens use electric fields to target roots.

Plant roots generate electrical currents and associated electrical fields as a consequence of electrogenic ion transport at the root surface. Here we demonstrate that the attraction of swimming zoospores of oomycete plant pathogens to plant roots is mediated in part by electrotaxis in natural root-generated electric fields. The zones of accumulation of anode- or cathode-seeking zoospores adjacent to intact and wounded root surfaces correlated with their in vitro electrotactic behavior. Manipulation of the root electrical field was reflected in changes in the pattern of zoospore accumulation and imposed focal electrical fields were capable of overriding endogenous signals at the root surface. The overall pattern of zoospore accumulation around roots was not affected by the presence of amino acids at concentrations expected within the rhizosphere, although higher concentrations induced encystment and reduced root targeting. The data suggest that electrical signals can augment or override chemical ones in mediating short-range tactic responses of oomycete zoospores at root surfaces.

Electricity↗