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Influx and efflux of p in roots of intact maize plants : double-labeling with p and p.

Rates of P influx and efflux were determined in whole plants at ambient P concentrations comparable to those found in soil solutions. Maize (Zea mays L. var NC+59) seedlings were trimmed (endosperm and adventitious roots removed) and grown in a greenhouse in solution cultures at P concentrations of approximately 0.4 and 1.8 micromolar. Roots of intact plants previously exposed to (32)P-labeled solutions at 0.2 and 2.0 micromolar P for 48 hours were rinsed 10 minutes in P-free solution and exposed to (33)P solutions at 0.2 and 2.0 micromolar for 10 minutes. Net depletion of (33)P from and appearance of (32)P in the ambient solution were used to measure influx and efflux. The ration of (32)P efflux to (33)P influx was about 0.68 at 0.2 micromolar and 0.08 at 2.0 micromolar. When plants were allowed to deplete P from solutions, the P concentration in the medium dropped to about 0.15 micromolar within 24 hours and 0.05 micromolar within 60 hours. Results indicate that P efflux is a substantial component of net P accumulation at P concentrations normally found in soil solutions.

Journal Article↗

Polyamines and Root Formation in Mung Bean Hypocotyl Cuttings : II. Incorporation of Precursors into Polyamines.

The incorporation of [(14)C]arginine and [(14)C]ornithine into various polyamines was studied in mung bean (Vigna radiata [L.] Wilczek) hypocotyl cuttings with respect to the effect of indole-3-butyric acid on adventitious root formation.Both [(14)C]arginine and [(14)C]ornithine are rapidly incorporated into putrescine, spermidine, and spermine, with similar kinetics, during 5- to 24-hour incubation periods. The incorporation of arginine into putrescine is generally higher than that of ornithine. The biosynthesis of putrescine and spermidine from the precursors, in the hypocotyls, is closely related to the pattern of root formation: a first peak at 0 to 24 hours corresponding to the period of root primordia development, and a second peak of putrescine biosynthesis at 48 to 72 hours corresponding to root growth and elongation. Indole-3-butyric acid considerably enhances putrescine biosynthesis in both phases, resulting in an increase of the putrescine/spermidine ratio.It is concluded that the promotive effect of indole-3-butyric acid on putrescine biosynthesis, from both arginine and ornithine, supports the hypothesis that auxin-induced root formation may require the promotion of polyamine biosynthesis.

Journal Article↗

The poplar root transcriptome: analysis of 7000 expressed sequence tags.

To date, most poplar expressed sequence tags (ESTs) are from above-ground tissues such as wood, leaf and buds. Here, we present a large-scale production of ESTs from roots of the hybrid cottonwood, Populus trichocarpaxdeltoides. cDNA libraries were generated from the root system of 2-month-old rooted cuttings, and roots of 2.5-month-old cuttings water-stressed for 19 days. Partial sequences obtained from 7013 clones were assembled into 1347 clusters and 3527 singletons. This set of ESTs represents 4874 unique transcripts expressed in roots. Putative functions could be assigned to 3021 (62%) of the transcripts. A significant portion of the ESTs encode proteins of common metabolic pathways; energy and metabolism represented 5% and 8% of total transcripts, respectively. Of specific interest to root functions are the 6% of ESTs involved in signalling pathways and hormone metabolism, and 4% encoding transporters and channels. The current poplar root ESTs and the aspen root ESTs present in public databases represent 6700 unique transcripts. The Unigene set was selected from the ESTs and used to generate nylon microarrays. Changes in aquaporins and transporter transcripts were then studied during adventitious root development.

Aquaporins↗

Plant steroid hormones.

After a brief survey of the literature on the recently discovered brassinolide, some of our results with corticosteroids are given. A number of structural requirements of corticosteroids were deduced in a bioassay involving elongation growth and lateral root formation. To explain the activity of glucocorticoids, the involvement of a receptor protein is suggested as well as an enhanced RNA synthesis. However, the structural requirements found are not applicable to experiments on adventitious root formation, implying a different mode of action of corticosteroids in this test system.

Adrenal Cortex Hormones↗

Role of Pseudomonas putida indoleacetic acid in development of the host plant root system.

Many plant-associated bacteria synthesize the phytohormone indoleacetic acid (IAA). While IAA produced by phytopathogenic bacteria, mainly by the indoleacetamide pathway, has been implicated in the induction of plant tumors, it is not clear whether IAA synthesized by beneficial bacteria, usually via the indolepyruvic acid pathway, is involved in plant growth promotion. To determine whether bacterial IAA enhances root development in host plants, the ipdc gene that encodes indolepyruvate decarboxylase, a key enzyme in the indolepyruvic acid pathway, was isolated from the plant growth-promoting bacterium Pseudomonas putida GR12-2 and an IAA-deficient mutant constructed by insertional mutagenesis. The canola seedling primary roots from seeds treated with wild-type P. putida GR12-2 were on average 35 to 50% longer than the roots from seeds treated with the IAA-deficient mutant and the roots from uninoculated seeds. In addition, exposing mung bean cuttings to high levels of IAA by soaking them in a suspension of the wild-type strain stimulated the formation of many, very small, adventitious roots. Formation of fewer roots was stimulated by treatment with the IAA-deficient mutant. These results suggest that bacterial IAA plays a major role in the development of the host plant root system.

Amino Acid Sequence↗

Soybean Roots Retain the Seed Urease Isozyme Synthesized during Embryo Development.

Roots of young soybean (Glycine max [L.] Merr.) plants (up to 25 days old) contain two distinct urease isozymes, which are separable by hydroxyapatite chromatography. These two urease species (URE1 and URE2) differ in: (a) electrophoretic mobility in native gels, (b) pH dependence, and (c) recognition by a monoclonal antibody specific for the seed ("embryo-specific") urease. By these parameters root URE1 urease is similar to the abundant embryo-specific urease isozyme, while root URE2 resembles the "ubiquitous" urease which has previously been found in all soybean tissues examined (leaf, embryo, seed coat, and cultured cells). The embryo-specific and ubiquitous urease isozymes are products of the Eu1 and Eu4 structural genes, respectively. Roots of the eu1-sun/eu1-sun genotype, which lacks the embryo-specific urease (i.e. ;seed urease-null'), contain no URE1 urease activity. Roots of eu4/eu4, which lacks ubiquitous urease, lack the URE2 (leaflike) urease activity. From these genetic and biochemical criteria, then, we conclude that URE1 and URE2 are the embryo-specific and ubiquitous ureases, respectively. Adventitious roots generated from cuttings of any urease genotype lack URE1 activity. In seedling roots the seedlike (URE1) activity declines during development. Roots of 3-week-old plants contain 5% of the total URE1 activity of the radicle of 4-day-old seedlings, which, in turn, has approximately the same urease activity level as the dormant embryonic axis. The embryo-specific urease incorporates label from [(35)S]methionine during embryo development but not during germination, indicating that there is no de novo synthesis of the embryo-specific (URE1) urease in the germinating root. We conclude that the seedlike urease (URE1) found in roots of young soybean plants is a remnant of the Eu1-encoded, abundant, embryo-specific urease which accumulates in the embryonic root axis during seed development.

Journal Article↗

Newly featured infection events in a supernodulating soybean mutant SS2-2 by Bradyrhizobium japonicum.

Supernodulating soybean (Glycine max L. Merr.) mutant SS2-2 and its wild-type counterpart, Sinpaldalkong 2, were examined for the microstructural events associated with nodule formation and development. SS2-2 produced a substantially higher percentage of curled root hairs than the wild type, especially at 14 days after inoculation with Bradyrhizobium japonicum. In addition, there was new evidence that in SS2-2, B. japonicum also entered through fissures created by the emerging adventitious root primordia. Early steps of nodule ontogeny were faster in SS2-2, and continued development of initiated nodules was more frequent and occurred at a higher frequency than in the wild type. These data suggest that the early expression of autoregulation is facilitated by decreasing the speed of cortical cell development, leading to the subsequent termination of less-developed nodules. The nodules of SS2-2 developed into spherical nodules like those formed on the wild type. In both the wild type and supernodulating mutant, vascular bundles bifurcate from root stele and branch off in the nodule cortex to surround the central infected zone. These findings indicate that SS2-2 has complete endosymbiosis and forms completely developed nodule vascular bundles like the wild type, but that the speed of nodule ontogeny differs between the wild type and SS2-2. Thus, SS2-2 has a novel symbiotic phenotype with regard to nodule organogenesis.

Bradyrhizobium↗

Origin and development in vitro of shoot buds and somatic embryos from intact roots of Helianthus annuus x H. tuberosus.

A variant clone of the tetraploid (2n = 4x = 68) interspecific hybrid Helianthus annuus x H. tuberosus derived by in vitro tissue culture showed a deviation from the usual pattern of organization of the plant body. This variant developed shoot-like structures and somatic embryos from intact adventitious roots of in vitro-grown plantlets. The morphogenetic structures were not normally able to differentiate complete plants. They did show cellular proliferation with the inception of additional secondary embryos, leaf-like structures and unorganized masses of callus. Nevertheless, some ectopic structures isolated from roots and transferred onto fresh basal medium without growth regulators were able to produce plantlets that exhibited the same phenotype as the original clone. Histological analyses demonstrate that they originate from cortical cells in association with the development of lateral root primordia.

Culture Techniques↗

MdWRKY75 interacts with MdWOX11 to modulate root growth under salt stress in apple.

The root system is pivotal for plant development, enabling both vegetative growth and tolerance to abiotic stresses like salinity. However, the molecular mechanisms governing root adaptive development in response to salt stress remain poorly understood in apple (Malus domestica Borkh.). In this study, we identified the salt stress-responsive WRKY transcription factor MdWRKY75. Overexpression of MdWRKY75 in transgenic apple negatively regulates adventitious root (AR) formation and salt stress tolerance, whereas reducing MdWRKY75 expression yields the opposite phenotype. Moreover, MdWRKY75 directly binds to the promoter of MdSAUR15 (SMALL AUXIN UP RNA15) and transcriptionally represses the expression of MdSAUR15, which, when overexpressed, promotes AR formation and enhances salt stress tolerance. We further demonstrated that MdWRKY75 interacts with MdWOX11, a WUSCHEL-related homeobox (WOX) transcription factor, both in vitro and in vivo. MdWOX11 expression is upregulated and enhances AR formation under salt stress. Additionally, MdWOX11 reduces the binding of MdWRKY75 to the MdSAUR15 promoter, and alleviates the MdWRKY75-mediated inhibitory effect on MdSAUR15 expression. Collectively, our study provides a MdWOX11-MdWRKY75-MdSAUR15 module regulating root adaptation in response to salt stress in apple.

Malus↗

Root-specific expression of genes for novel glycine-rich proteins cloned by use of an antiserum against xylem sap proteins of cucumber.

Candidate cDNAs for xylem sap proteins (XSPs) were isolated by screening of a cucumber cDNA library prepared from poly(A)+ RNA of cucumber roots with an antiserum raised against the XSPs of cucumber. The patterns of expression of corresponding transcripts and the sequences of the cDNAs were examined. Two cDNAs with a glycine-rich domain in each deduced amino acid sequence and whose transcripts were expressed predominantly in roots were selected for further analysis. Both cDNAs encoded proteins with a putative signal sequence, with similar non-glycine-rich domains at the amino terminal of the encoded polypeptides. The corresponding mRNAs accumulated concurrently with the formation of adventitious roots from cuttings of cucumber hypocotyls and the development of vascular tissues, and strong expression of the mRNAs was detected in the root-hair zone of tap roots. These data suggest that the production of the two novel glycine-rich proteins, which are putative XSPs, might be associated with the functions of the vascular tissues in roots.

Amino Acid Sequence↗

Decline in stomatal response to leaf water deficit in Corylus maxima cuttings.

Many woody species can be propagated from leafy cuttings. However, following rooting, cuttings of Corylus maxima Mill. cv. Purpurea do not always survive the transition from a highly supportive rooting environment (e.g., fog) to a more natural environment where evaporative demand is higher. We found that it is not the supply of water to leaves, but stomatal dysfunction that leads to severe water deficits in the rooted cuttings. Two hours after well-rooted cuttings were transferred from the rooting environment, we were able to relate visible signs of leaf water deficit to high stomatal conductance (g(s)) and low relative water content (R). Small expanding leaves (L3) had unusually high g(s) and lower R than fully expanded leaves (L1). Although high cuticular conductances (g(c)) were occasionally observed in L3, SEM confirmed that increased total leaf conductance (g) was mainly a result of abnormally wide stomatal opening. We measured changes in the ability of stomata to control water loss during rooting by determining stomatal responsiveness to leaf water deficit in detached L1 and L3 harvested from cuttings during the first 75 days after severance from stock plants. Reduced stomatal responsiveness was observed within 7 days of severance, prior to adventitious root formation, and was more pronounced in L3 than in L1. A period of acclimatization after rooting (no leaf wetting, but a vapor pressure deficit of 0.20 kPa) reduced g(s) by 50% in L3 but not in L1, and partially restored stomatal responsiveness in L1 but not in L3. After rooting, the original leaves on the cutting retained substantial capacity for photosynthesis (e.g., in L1, 8 micromol m(-2) s(-1) at a photosynthetic photon flux density of 400 micromol m(-2) s(-1)). The implications of the results for post-rooting acclimatization procedures are discussed.

Betulaceae↗

Influence of 2,3,5-Triiodobenzoic Acid and 1-N-Naphthylphthalamic Acid on Indoleacetic Acid Transport in Carnation Cuttings: Relationship with Rooting.

(3)H-IAA transport in excised sections of carnation cuttings was studied by using two receiver systems for recovery of transported radioactivity: agar blocks (A) and wells containing a buffer solution (B). When receivers were periodically renewed, transport continued for up to 8 h and ceased before 24 h. If receivers were not renewed, IAA transport decreased drastically due to immobilization in the base of the sections. TIBA was as effective as NPA in inhibiting the basipetal transport irrespective of the application site (the basal or the apical side of sections). The polarity of IAA transport was determined by measuring the polar ratio (basipetal/acropetal) and the inhibition caused by TIBA or NPA. The polar ratio varied with receiver, whereas the inhibition by TIBA or NPA was similar. Distribution of immobilized radioactivity along the sections after a transport period of 24 h showed that the application of TIBA to the apical side or NPA to the basal side of sections, increased the radioactivity in zones further from the application site, which agrees with a basipetal and acropetal movement of TIBA and NPA, respectively. The existence of a slow acropetal movement of the inhibitor was confirmed by using (3)H-NPA. From the results obtained, a methodological approach is proposed to measure the variations in polar auxin transport. This method was used to investigate whether the variations in rooting observed during the cold storage of cuttings might be related to changes in polar auxin transport. As the storage period increased, a decrease in intensity and polarity of auxin transport occurred, which was accompanied by a delay in the formation and growth of adventitious roots, confirming the involvement of polar auxin transport in supplying the auxin for rooting.

Journal Article↗

Evidence for symplastic involvement in the radial movement of calcium in onion roots.

The pathway of Ca(2+) movement from the soil solution into the stele of the root is not known with certainty despite a considerable body of literature on the subject. Does this ion cross an intact, mature exodermis and endodermis? If so, is its movement through these layers primarily apoplastic or symplastic? These questions were addressed using onion (Allium cepa) adventitious roots lacking laterals. Radioactive Ca(2+) applied to the root tip was not transported to the remainder of the plant, indicating that this ion cannot be supplied to the shoot through this region where the exodermis and endodermis are immature. A more mature zone, in which the endodermal Casparian band was present, delivered 2.67 nmol of Ca(2+) mm(-1) treated root length d(-1) to the transpiration stream, demonstrating that the ion had moved through an intact endodermis. Farther from the root tip, a third zone in which Casparian bands were present in the exodermis as well as the endodermis delivered 0.87 nmol Ca(2+) mm(-1) root length d(-1) to the transpiration stream, proving that the ion had moved through an unbroken exodermis. Compartmental elution analyses indicated that Ca(2+) had not diffused through the Casparian bands of the exodermis, and inhibitor studies using La(3+) and vanadate (VO(4)(3-)) pointed to a major involvement of the symplast in the radial transport of Ca(2+) through the endodermis. It was concluded that in onion roots, the radial movement of Ca(2+) through the exodermis and endodermis is primarily symplastic.

Biological Transport↗

Influence of (22S,23S)-homobrassinolide on rooting capacity and survival of adult Norway spruce cuttings.

To determine whether brassinosteroids promote clonal propagation of mature coniferous trees, cuttings of adult Norway spruce (Picea abies (L.) Karst.) trees were harvested in late March, treated with (22S,23S)-28-homobrassinolide (SSHB), and stored at 2 degrees C in darkness until they were planted in May. Treatment of the cuttings with SSHB significantly enhanced the formation of adventitious roots by increasing percent rooting from 50% in the control cuttings to 92% in the 60 ppm SSHB treatment. N(6)-Benzyladenine (BA) stimulated rooting slightly although not significantly, but it counteracted the stimulative effect of SSHB when cuttings were treated with both compounds together. In a second study, the above-ground parts of medium-sized mature clonal Norway spruce trees were pretreated from the end of June till August with SSHB or BA or both. Needle retention of cuttings taken from the pretreated trees the following spring was prolonged until September and the cuttings flushed, but did not root, whereas cuttings from untreated stock plants died before September. No interaction between BA and SSHB was observed. When these pretreatments were repeated the following year on the same mature clonal trees rooting of cuttings was significantly improved in the 5 ppm SSHB treatment.

Journal Article↗

Laboratory-scale measurements of N2O and CH4 emissions from hybrid poplars (Populus deltoides x Populus nigra).

The purpose of this study was to determine whether or not young hybrid poplar (Populus deltoides x Populus nigra) could transport landfill biogas internally from the root zone to the atmosphere, thereby acting as conduits for landfill gas release. Fluxes of methane (CH4) and nitrous oxide (N2O) from the seedlings to the atmosphere were measured under controlled conditions using dynamic flux chambers and a tunable diode laser trace gas analyser (TDLTGA). Nitrous oxide was emitted from the seedlings, but only when extremely high soil N2O concentrations were applied to the root zone. In contrast, no detectable emissions of CH4 were measured in a similar experimental trial. Visible plant morphological responses, characteristic of flood-tolerant trees attempting to cope with the negative effects of soil hypoxia, were observed during the CH4 experiments. Leaf chlorosis, leaf abscission and adventitious roots were all visible plant responses. In addition, seedling survival was observed to be highest in the biogas 'hot spot' areas of a local municipal solid waste landfill involved in this study. Based on the available literature, these observations suggest that CH4 can be transported internally by Populus deltoides x Populus nigra seedlings in trace amounts, although future research is required to fully test this hypothesis.

Air Pollutants↗

Light sources with different spectra affect root and mycorrhiza formation in Scots pine in vitro.

We studied the effects of broad-spectrum light quality on the interaction between the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker and Couch and Scots pine (Pinus sylvestris L.) seedlings and hypocotyl cuttings cultured in vitro. The light sources were cool white (CW), warm white (WW) and red-rich daylight (RD) fluorescent lamps. Inoculation with P. tinctorius enhanced adventitious root formation of the cuttings in all light treatments. Rooting of the inoculated cuttings was highest in WW light (89%), followed by CW (73%) and RD light (66%). During 6 weeks of in vitro culture, rooted cuttings formed only a few lateral roots. The fungus grew over lateral roots, but the Hartig net was absent in all light treatments. In non-inoculated cuttings, neither root formation nor subsequent root growth was affected by light quality. In the seedling experiment, inoculation in the WW treatment resulted in a significantly (P < 0.05) greater number of lateral roots than inoculation in the RD treatment. The percentage of lateral roots covered with fungal hyphae was also highest in WW light (62%), followed by CW (50%) and RD (27%) light. A similar pattern was observed in the intensity of Hartig net formation. We conclude that effects of broad-spectrum light quality on the ectomycorrhizal fungus-root interaction are dependent on the developmental stage of the root.

Basidiomycota↗

Quantitative determination of secoiridoid and gamma-pyrone compounds in Gentiana lutea cultured in vitro.

The production of secondary metabolites was studied in shoots, roots, and hairy roots of Gentiana lutea obtained in vitro. In shoots, both secoiridoid and gamma-pyrone compounds were detected in amounts similar to those found in aerial parts of plants collected from nature. The most abundant secoiridoid was gentiopicrin while mangiferin was the main compound among the gamma-pyrones. The adventitious roots obtained in vitro showed a poor biosynthetic capacity. Upon infection with Agrobacterium rhizogenes, nine hairy root clones were established which differed in the amount of secondary metabolites.

Chromatography, High Pressure Liquid↗

Genetic analysis of rooting ability of transplanted rice (Oryza sativa L.) under different water conditions.

In order to assess the benefits of superior rooting ability of rice (Oryza sativa L.) for growth after transplanting under water-limiting conditions, genetic differences in the rooting ability of rice seedlings 30 d after sowing, with their visible roots either pruned or not pruned, were quantified by several root parameters 4 d after transplanting (DAT), under flooded or non-flooded paddy fields (four treatments in total), together with production traits at maturity. Ninety-eight recombinant inbred lines from the two japonica ecotypes, a lowland variety Otomemochi, and an upland variety Yumenohatamochi, were genotyped with 107 simple sequence repeat (SSR) markers. Otomemochi in general produced more adventitious roots, partitioned a greater proportion of biomass to roots, and had a greater increment of root dry weight (DeltaRW) at 4 DAT than Yumenohatamochi, but these variety differences were less clear under non-flooded conditions without root pruning. Several quantitative trait loci (QTLs) associated with rooting ability were identified mainly in chromosomes 1, 4, and 6 across the four treatments or in combined analysis. On the other hand, QTLs for DeltaRW around RM2357 in chromosome 5 and for maximum new root length in RM215-RM205 in chromosome 9 were found only under root pruning treatments and under non-flooded conditions without root pruning, respectively. Greater DeltaRW was associated with higher head dry weight per hill at maturity only in the non-flooded and root-pruning treatments. This study suggests the importance of rooting ability after transplanting and, possibly, other mechanisms for adaptation to non-flooded conditions.

Oryza↗