Hormone interactions at the root apical meristem

Plants exhibit an amazing developmental flexibility. Plant embryogenesis results in the establishment of a simple apical-basal axis represented by apical shoot and basal root meristems. Later, during postembryonic growth, shaping of the plant body continues by the formation and activation of numerous adjacent meristems that give rise to lateral shoot branches, leaves, flowers, or lateral roots. This developmental plasticity reflects an important feature of the plant's life strategy based on the rapid reaction to different environmental stimuli, such as temperature fluctuations, availability of nutrients, light or water and response resulting in modulation of developmental programs. Plant hormones are important endogenous factors for the integration of these environmental inputs and regulation of plant development. After a period of studies focused primarily on single hormonal pathways that enabled us to understand the hormone perception and signal transduction mechanisms, it became obvious that the developmental output mediated by a single hormonal pathway is largely modified through a whole network of interactions with other hormonal pathways. In this review, we will summarize recent knowledge on hormonal networks that regulate the development and growth of root with focus on the hormonal interactions that shape the root apical meristem.     

Mitotic cycle kinetics of root meristems of isolated barley embryos and intact seedlings. Labelling of nuclei by3H- thymidine and its cytogenetic consequences

The duration of the mitotic cycle and its individual phases was estimated in root meristems of isolated barley embryos and intact barley seedlings by means of pulse labelling with³H-thymidine and construction of labelled mitoses curve. The duration of the whole mitotic cycle in the cell population of root meristems of isolated barley embryos cultivated in the aerated liquid complete medium is 12.2 h. The mitotic cycle time of root meristems of intact barley seedlings, oultived in Petri dishes on wet blotting paper is 9.2 h. Most of root meristem cells belong to the fraction of rapidly proliferating cells, but this fraction exerts a high degree of variability by itself. Pulse treatment by³H-thymidine in our experimental conditions (74 kBq ml^-1 - or 2 μCi ml^-1, exposure 0.5 h) did not induoe any chromosomal aberrations in unlabelled cells and only a very low frequency of chromosomal aberrations in labelled cells. Measuring the cell population kinetics by pulse labelling with³H-thymidine can be used simultaneously with the study of induction of ohromosomal aberrations by mutagens.     

Short term cultivation of isolated barley roots and their mitotic activity

Roots were excised from barley embryos cultivated in the complete liquid nutrient solution and cultivated in the same nutrient solution separately. The excised roots continued their growth but a progressive decrease in the growth rate was observed. There was a considerable short-term drop of the mitotic activity immediately after excision, which was followed by a compensatory increase and then equilibrium was reached 12 h after excision. During the next at least three days, the mitotic index of isolated barley roots varied between 5–6.5%, which is slightly lower than the mitotic index of the root meristems of isolated barley embryos under identical conditions. The mitotic cycle index of isolated barley roots and the size of the root meristem later decreased gradually.     

Possibility of synchronizing cell populations in embryonic barley meristem

Cell populations of the apical root parts, stem embryo and the leaf of barley seedlings are found to have different sensitivity to the synchronizing effect of 5-aminouracil, low temperature (+2 degrees C) and colchicine. The effect of 5-aminouracil and low temperature in the presence of colchicine proved to be the most effective in respect to synchronization of the root meristem cell populations. It also increases significantly the mitotic activity in the stem embryo and leaf meristems. The leaf meristem is more sensitive to low temperature as compared to the stem embryo meristem.     

Soil structure and plant growth: Impact of bulk density and biopores

Compacted soils are not uniformly hard; they usually contain structural cracks and biopores, the continuous large pores that are formed by soil fauna and by roots of previous crops. Roots growing in compacted soils can traverse otherwise impenetrable soil by using biopores and cracks and thus gain access to a larger reservoir of water and nutrients. Experiments were conducted in a growth chamber to determine the plant response to a range of uniform soil densities, and the effect of artificial and naturally-formed biopores. Barley plants grew best at an intermediate bulk density, which presumably represented a compromise between soil which was soft enough to allow good root development but sufficiently compact to give good root-soil contact. Artificial 3.2 mm diameter biopores made in hard soil gave roots access to the full depth of the pot and were occupied by roots more frequently than expected by chance alone. This resulted in increased plant growth in experiments where the soil was allowed to dry. Our experiments suggest that large biopores were not a favourable environment for roots in wet soil; barley plants grew better in pots containing a network of narrow biopores made by lucerne and ryegrass roots, responded positively to biopores being filled with peat, and some pea radicles died in biopores. A theoretical analysis of water uptake gave little support to the hypothesis that water supply to the leaves was limiting in either very hard or very soft soil. The net effect of biopores to the plant would be the benefits of securing extra water and nutrients from depth, offset by problems related to poor root-soil contact in the biopore and impeded laterals in the compacted biopore walls.     

Growth-Regulating Substances and the Growth of Tiller Buds in Barley; Effects of Cytokinins

An hypothesis was set up from which it was predicted that applicationof cytokinin to barley seedlings grown without mineral nutrientswould lead to rapid growth of the coleoptile and first leaftiller buds. Application of cytokinins to the leaves was ineffective,but supplying a number of known cytokinins by steeping the rootsof 4 d old seedlings in solution for 4 h led to significantgrowth of the coleoptile bud. Adenine and cytokinin analogueshad no effect. Supplying cytokinins through the roots also furtherenhanced the growth of buds of plants given mineral nutrients.Cytokinin treatment reduced root dry matter, with small reductionsin mean axis length and number of lateral roots. For plantsnot given mineral nutrients reduction in root weight was compensatedby an increase in weight of the aerial parts; however, for plantssupplied with mineral nutrients this was not so and the lowerroot weight resulted in a smaller total plant dry weight. An interpretation of tiller bud growth in terms of control byinteracting effects of mineral nutrition, assimilate supply,and cytokinin availability is proposed.     

Abscisic acid rescues the root meristem defects of the Medicago truncatula latd mutant

The LATD gene of the model legume, Medicago truncatula, is required for the normal function of three meristems, i.e. the primary root, lateral roots and nitrogen-fixing nodules. In latd mutants, primary root growth eventually arrests, resulting in a disorganized root tip lacking a presumptive meristem and root cap columella cells. Lateral root organs are more severely affected; latd lateral roots and nodules arrest immediately after emerging from the primary root, and reveal a lack of organization. Here we show that the plant hormone, abscisic acid (ABA), can rescue the latd root, but not nodule, meristem defects. Growth on ABA is sufficient to restore formation of small, cytoplasm-rich cells in the presumptive meristem region, rescue meristem organization and root growth and formation of root cap columella cells. In contrast, inhibition of ethylene synthesis or signaling fails to restore latd primary root growth. We find that latd mutants have normal levels of ABA, but exhibit reduced sensitivity to the hormone in two other ABA-dependent processes: seed germination and stomatal closure. Together, these observations demonstrate that the latd mutant is defective in the ABA response and indicate a role for LATD-dependent ABA signaling in M. truncatula root meristem function.     

Phosphorus acquisition from soil by white lupin (Lupinus albus L.) and soybean (Glycine max L.), species with contrasting root development

White lupin and soybean have contrasting root morphologies: white lupin develops proteoid or cluster roots, roots with discreet clusters of short, determinate branch roots (rootlets) while soybean develops a more fibrous root system with evenly distributed, longer branch roots. Growth and P acquisition by white lupin and soybean were compared in a soil high in bound, total P, with or without additional inorganic P applied in solution. Additional P increased biomass by 25% and doubled total P in soybean. In contrast, white lupin did not respond to additional P in biomass or total P. However added P decreased cluster development on proteoid roots indicating that white lupin sensed the added P. The reduction in cluster weight per plant was exactly countered by an increase in dry weight of other roots. Soybean root development responded to P application, proliferating branch roots with active meristems in the upper portion of the soil profile where P was applied, and reducing root weight to plant weight by 13%. White lupin did not proliferate roots in response to P application. When P was not added to soil, soybean and lupin acquired similar P per unit root dry weight. However, white lupin accumulated 4.8 times more P per unit root length, suggesting that P acquisition in these plants involved other mechanisms such as the exudation of P solubilizing compounds. Soybean accessed P by developing more root length thus colonising more soil volume than white lupin and, therefore, was better able to take advantage of the added P. Pericycle and root tip meristem activities were critical to the differences in root development between white lupin and soybean, and therefore their responses to plant and soil P.     

The involvement of wheat and common bean lectins in the control of cell division in the root apical meristems of various plant species

The effects of wheat germ agglutinin (WGA) and phytohemagglutinin (PHA) at the concentration of 1 mg/l on the rate of cell division in the root apical meristem of wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), rice (Oryza sativa L.), and common bean (Phaseolus vulgaris L.) seedlings were compared. WGA enhanced cell division in the roots of barley and rice approximately similarly as in wheat roots but did not affect division of meristematic cells in the roots of common bean seedlings. In contrast PGA enhanced mitotic activity in the root apical meristem of common bean seedlings but did not affect division in the wheat and barley roots. Seedling treatment with lectins shifted the hormonal balance in them toward accumulation of growth activators (IAA and cytokinins). The relationship between lectin and hormonal systems in the control of cell division is discussed.     

Nutrient uptake and growth of barley as affected by soil compaction

A field experiment with different levels of compaction was carried out on a mouldboard ploughed silty clay, with the objective of studying the effects on plant nutrient uptake and growth. Soil from the field was also used in laboratory studies of carbon and nitrogen mineralization, and plant uptake of water and nutrients. In the field, low as well as high bulk densities reduced biomass production and nutrient uptake of barley (Hordeum vulgare L.) compared to intermediate bulk densities, where grain yield was approximately 20% higher. In the beginning of the growing season, the concentration of phosphorus and potassium was lowest in plants grown in the loosest and in the most compacted soil, and suboptimal for plant growth. The uptake of nutrients transported by diffusion was more affected by compaction than for nutrients transported by mass flow. The reasons for lowered uptake in loose compared to moderately compacted soil could be reduced root-to-soil contact, a low diffusion coefficient for nutrients and/or reduced mass transport of water to seed and roots. Differences in plant nutrient concentrations between treatments gradually declined until harvest. Immediately after compaction there was probably oxygen deficiency in the compacted soil since the air-filled porosity was critically low, but as the soil dried out, mechanical resistance to root growth may have become a more important growth-limiting factor. In the laboratory study, severe compaction reduced carbon mineralization and uptake of water and nutrients by roots, and caused denitrification. There were only small differences between loose and moderately compacted soil in carbon mineralization, nitrogen concentration in the soil, uptake of water and nutrients and dry matter yield. The large yield increase due to recompaction in the field was not reproduced in the laboratory. Possible reasons are differences in soil temperature between the field and laboratory, in the sowing and fertilizing methods, the pretreatment of the soil and in the spatial variability of bulk density. It is possible that recompaction is needed only in the uppermost part of the soil, which is the loosest, dries out first, and is where the seed as well as the fertilizer are placed. This revised version was published online in July 2006 with corrections to the Cover Date.     

Activation of cell division in the quiescent center of excised maize root tip

The phenomenon of activation of cell proliferation in the quiescent center of excised maize roots is described. The root tips were grown on wet filter paper in Petri dishes. This phenomenon was observed in 8 to 14 maize cultivars and was absent in excised Arabidopsis root tips. The distribution of mitoses in meristems greatly varied in roots of individual seedlings from the same seed lot and seedlings of different cultivars. Meristem opening was observed after the removal of small root tips not longer than 3 mm and intact seminal roots. Sucrose (2%) and 10(-6)-10(-8) M indole-3-acetic acid did not prevent meristem opening. These findings indicate that the state of quiescent center is maintained by a system of intercellular and interorgan relations, which are to be clarified.     

Mapping meristem respiration of Prunus persica (L.) Batsch seedlings: potential respiration of the meristems, O2 diffusional constraints and combined effects on root growth

Root system architecture partially results from meristem activities, which themselves depend on endogenous and environmental factors, such as O2 depletion. In this study, meristem respiration and growth was measured in the root systems of three Prunus persica (L.) Batsch seedlings. The spatial distribution of meristem respiration within the root system was described, and the relationship between the respiration rates and meristem radii was analysed, using a model of radial O2 diffusion and consumption within the root. Histological observations were also used to help interpret the results. Respiration rates were linearly correlated to the root growth rates (rho 2 = 0.9). Respiration reached values greater than 3.5 x 10(-13) mol O2 s-1 for active meristems. The taproot meristem consumed more O2 than the rest of the entire root system meristems. Similarly, the first order lateral meristems used more O2 than the second order ones. A near hyperbolic relationship between respiration rates and meristem radii was observed. This can be explained by a model of radial O2 diffusion and consumption within the root. Therefore, only one maximum potential respiration rate and one O2 diffusion coefficient was estimated for all the meristems.     

Response to auxin changes during maturation-related loss of adventitious rooting competence in loblolly pine (Pinus taeda) stem cuttings

Hypocotyl cuttings (from 20- and 50-day-old Pinus taeda L. seedlings) rooted readily within 30 days in response to exogenous auxin, while epicotyl cuttings (from 50-day-old seedlings) rarely formed roots within 60 days. Responses to auxin during adventitious rooting included the induction of cell reorganization and cell division, followed by the organization of the root meristem. Explants from the bases of both epicotyl and hypocotyl cuttings readily formed callus tissue in response to a variety of auxins, but did not organize root meristems. Auxin-induced cell division was observed in the cambial region within 4 days, and later spread to the outer cortex at the same rate in both tissues. Cells at locations that would normally form roots in foliated hypocotyl cuttings did not produce callus any differently than those in other parts of the cortex. Therefore, auxin-induced root meristem organization appeared to occur independently of auxin-induced cell reorganization/division. The observation that N-(1-naphthyl)phthalamic acid (NPA) promoted cellular reorganization and callus formation but delayed rooting implies the existence of an auxin signal transduction pathway that is specific to root meristem organization. Attempts to induce root formation in callus or explants without foliage were unsuccessful. Both the cotyledon and epicotyl foliage provided a light-dependent product other than auxin that promoted root meristem formation in hypocotyl cuttings.     

Neutron imaging reveals internal plant water dynamics

Background and aims

Knowledge of plant water fluxes is critical for assessing mechanistic processes linked to biogeochemical cycles, yet resolving root water transport dynamics has been a particularly daunting task. Our objectives were to demonstrate the ability to non-invasively monitor individual root functionality and water fluxes within Zea mays L. (maize) and Panicum virgatum L. (switchgrass) seedlings using neutron imaging.

Methods

Seedlings were propagated for 1–3 weeks in aluminum chambers containing sand. Pulses of water or deuterium oxide were then tracked through the root systems by collecting consecutive radiographs during exposure to a cold-neutron source. Water flux was manipulated by cycling on a growth lamp to alter foliar demand for water.

Results

Neutron radiography readily illuminated root structure, root growth, and relative plant and soil water content. After irrigation there was rapid root water uptake from the newly wetted soil, followed by hydraulic redistribution of water through the root system to roots terminating in dry soil. Water flux within individual roots responded differentially to foliar illumination based on supply and demand of water within the root system.

Conclusions

Sub-millimeter scale image resolution revealed timing and magnitudes of root water uptake, redistribution within the roots, and root-shoot hydraulic linkages—relationships not well characterized by other techniques.     

Study of the effect of volatile substances from cereal roots

We traced the liberation and biological effect of volatile substances released from the roots of cereals,i. e. barley, wheat, rye and oats, on seedlings of the same and other plant species. Experiments were carried out in a closed glass apparatus with a static or circulating atmosphere in which the CO₂ and O₂ were permanently absorbed and supplemented, respectively. In some experiments the air was bubbled through water or through solutions of boric acid, barium hydroxide and potassium permanganate. The roots of all four cereals tested released volatile substances with a biological activity which appeared to be non-specific with respect to plant species. The effect of volatile substances was partially decreased by bubbling through water, barium hydroxide and boric acid and was completely removed after passing through the solution of potassium permanganate. Volatile substances liberated from roots of barley inhibited elongation of roots and coleoptile, decreased SH-group content and caused excessive formation of root hairs as well as inhibition of both dry matter production and respiration of roots of rye seedlings. Ethylene was found in the atmosphere of experimental vessels.     

Comparative anatomy of root meristem and root cap in some species of Podostemaceae and the evolution of root dorsiventrality

In the unusual aquatic Podostemaceae, the root is the leading organ of the plant body and is variously compressed and submerged as it adheres to rock surfaces in rapid water. In an anatomical comparison of the root apical meristems and root caps of 33 species that represent the major lineages of the family, the dorsiventrality of root meristems varied and was classified into four patterns: (1) The root cap is produced outward from a nearly radially symmetrical meristem. (2) The meristem and root cap are markedly dorsiventral; the outermost cells of the hood-shaped cap are acroscopic derivatives from bifacial initials on the ventral side, while the pattern on the dorsal side is similar to pattern 1. (3) Bifacial initials are on both the dorsal and ventral sides. (4) No root cap is present. An evolutionary polarity may be evident from pattern 1 to 2 and then to 3. Pattern 2 arose in the early evolution of the subfamily Podostemoideae and subsequently, pattern 3 arose in species with crustose roots, while the least specialized pattern 1 is retained in Tristichoideae and Weddellinoideae. Pattern 4 characterized by caplessness may have appeared recurrently in Tristichoideae and Podostemoideae. These evolutionary changes in the meristem preceded the specialization of external root morphologies.     

IS TRIGONELLINE A PLANT HORMONE?

Experimental results are presented so that trigonelline may be evaluated as a plant hormone. Trigonelline promotes preferential cell arrest in G2 of the cell cycle for about 40% of the cell population in root meristems of Pisum sativum. Trigonelline is present in ungerminated seeds and is transported from cotyledons to other tissues during early seedling development. These experimental results show that trigonelline satisfies all six criteria that have been used to establish whether a substance is a hormone. As the seedlings age from day 3 to 10, the concentration of trigonelline in meristems decreases and so does the proportion of cells arrested in G2. Trigonelline may be isolated from excised cotyledons and can be added back to decotyledonized seedlings or excised root meristems to have the same effect as found in intact organisms. Predominant cell arrest in G2 occurs in roots of some plant species, although other species show preponderant cell arrest in G1. Many members of the Preiss-Handler metabolic pathway show some ability to promote cell arrest in G2 but only at concentrations 100 (10^-5 m ) or 1,000 (10^-4 m ) times the concentration of trigonelline (10^-7 m ) necessary for function. The proportion of cells arrested in G2 is highly correlated with the concentration of trigonelline within the root meristem.     

Effect of associative bacteria on element composition of barley seedlings grown in solution culture at toxic cadmium concentrations

The response of barley seedlings to inoculation with associative rhizobacteria Azospirillum lipoferum 137, Arthrobacter mysorens 7, Agrobacterium radiobacter 10 and Flavobacterium sp. L30 was studied in hydroponic and quartz sand cultures in the presence of 50 microM CdCl2. Cadmium caused severe inhibition in the growth and uptake of nutrient elements by the plants. Inoculation with the bacteria slightly stimulated root length and biomass of hydroponically grown Cd-treated seedlings. The bacteria increased the content of nutrients such as P, Mg, Ca, Fe, Mn and Na in roots and or shoots of the plants grown in the absence of Cd. Positive changes in the element composition caused by the bacteria were less pronounced in Cd-treated plants, whereas the total amount of nutrients taken by the inoculated plants was generally increased significantly. The content of Cd in the inoculated plants was unchanged, except increased in roots upon addition of A. lipoferum 137. Inoculation did not affect the activity of peroxidase, alpha-mannosidase, phosphodiesterae, alpha-galactosidase, and concentration of sulfhydryl compounds used as biochemical markers of stress in plant roots. The results showed that associative bacteria were capable of decreasing partially the toxicity of Cd for the barley plants through the improvement in uptake of nutrient elements.     

DNA synthesis during the early stage of germination in the barley embryo meristems and its inhibition by N-methyl-N-nitrosourea

3 peaks of DNA synthesis were observed in the barley embryo of seeds, germinating for 49 h in running tap water at 25°C. The first peak, found after 22h, was formed by S-cells in the roots and in the 1st leaf meristem. The second peak (after 34–37h) and third peak (after 46–49 h) represents the S-cells in the roots, apex and 1st, 2nd and 3rd leaf meristems. Application of N-methyl-N-nitrosourea for 3 h at the onset of germination inhibited the rate of DNA synthesis and postponed the peaks of DNA synthesis in individual meristems of the embryo.