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PubMed2006

[Effects of irradiance on boron requirement of phaseolus aureus cutting for its adventitious root development].

This paper studied the boron requirement of Phaseolus aureus cutting for its adventitious root development under different photosynthetic active radiation (PAR). The results showed that exogenous boron supply was necessary for the adventitious root development of the cutting under 10 days of PAR at 50 or 100 micromol x m(-2) x s(-1), but unnecessary under 6 days of this irradiance and followed by 4 days of darkness. Exogenous boron supply was also necessary for the rooting under 6 days of PAR at 180 micromol x m(-2) x s(-1) and followed by 4 days of darkness. When the PAR at 100 micromol x m(-2) x s(-1) was employed for 10 days, the amount of soluble boron in each hypocotyl of the cutting was only half of that under 6 days of this irradiance and followed by 4 days of darkness, which might be the reason of the necessity of exogenous boron supply for the rooting under 10 days of PAR at 100 micromol x m(-2) x s(-1).

Boron↗
PubMed1995

LRP1, a gene expressed in lateral and adventitious root primordia of arabidopsis.

We describe a gene that is expressed in lateral and adventitious root primordia of Arabidopsis. The gene was identified by expression of a transposon-borne promoterless beta-glucuronidase gene in lateral root primordia. The gene, designated LRP1 for lateral root primordium 1, and its corresponding cDNA were cloned and sequenced. The expression pattern of the gene in lateral root primordia was confirmed by in situ hybridization with LRP1 cDNA probes. The LRP1 gene encodes a novel protein. LRP1 expression is activated during the early stages of root primordium development and is turned off prior to the emergence of lateral roots from the parent root. Insertion of the transposon in the LRP1 gene disrupted its expression. To evaluate the homozygous insertion line for a mutant phenotype, several aspects of wild-type lateral root development were analyzed. A mutant phenotype has not yet been identified in the insertion line; however, there is evidence that the gene belongs to a small gene family. LRP1 provides a molecular marker to study the early stages of lateral and adventitious root primordium development.

Amino Acid Sequence↗
PubMed2004

Adventitious root growth and ginsenoside accumulation in Panax ginseng cultures as affected by methyl jasmonate.

Adventitious roots of ginseng were treated with methyl jasmonate (MJ) up to 150 microM and cultured for 40 days. Up to 100 microM MJ inhibited the root growth but increase ginsenoside accumulation. In a two-stage bioreactor culture, total ginsenosides, after elicitation with 100 microM MJ peaked after 10 days at 48 mg g(-1) dry wt and then dropped sharply. Of the two groups of ginsenosides (Rb and Rg), higher amounts of Rb accumulated in the adventitious roots.

Acetates↗
PubMed2001

Ellagic acid formation from galloylglucoses by a crude enzyme of Cornus capitata adventitious roots.

The aqueous extract of acetone powder, which had been prepared from Cornus capitata 'Mountain Moon' adventitious roots, cultured in MS medium with a high concentration of Cu2+(10 microM), showed strong oxidative activity toward galloylglucoses. A compound formed from galloyglucoses, such as 1,2,3,4,6-penta-O-galloyl-beta-D-glucose and tannic acid, by the reaction with the crude enzyme solution of the adventitious roots was isolated and characterized as ellagic acid by spectrometric analyses.

Chromatography, High Pressure Liquid↗
PubMed2004

Adventitious root formation in Arabidopsis thaliana thin cell layers.

This paper describes, for the first time, de novo adventitious root formation from thin cell layers (TCLs) of Arabidopsis thaliana. The objective of the study was to determine the optimal hormonal and light conditions and the optimal exogenous Ca2+ concentration for obtaining adventitious rooting (AR) from A. thaliana TCLs and to identify the tissue(s) involved in the process. The results show that maximum AR was obtained with a single-phase method in the presence of 10 microM indole-3-butyric acid and 0.1 microM kinetin under continuous darkness for 30 days and with 0.6 mM exogenous CaCl2. The endodermis was the only tissue involved in root meristemoid formation. The role of Ca2+ in AR and the importance of using Arabidopsis TCLs in studies on the genetic/biochemical control of AR are discussed.

Adenine↗
PubMed2004

[Cloning of cDNA fragments related to adventitious root formation from mango cotyledon section].

Two cut surfaces of mango cotyledon (distal and proximal cut surfaces) showed different capability of adventitious root formation, only proximal cut surface could be induced to form the roots and the distal cut surface did not. cDNA fragments related to adventitious root formation from the cut sections were isolated with suppressive subtractive hybridization. The forward substracted cDNA library was constructed using the cDNAs of distal (non-rooting) cut surface as driver and the cDNAs of proximal (rooting) cut surface as tester. Six positive clones were obtained by Virtual Northern blots. In this study, the putative up-regulated genes showed by sequence analysis were reported in mango for the first time, the deduced proteins among the positive clones were homologous to transporters, transcriptional regulators and enzymes.

Blotting, Northern↗
PubMed2001

Cytokinin-like activity of N,N'-diphenylureas. N,N'-bis-(2,3-methylenedioxyphenyl)urea and N,N'-bis-(3,4-methylenedioxyphenyl)urea enhance adventitious root formation in apple rootstock M26 (Malus pumila Mill.).

Vegetative propagation of cuttings is a widespread method to multiplicate plants. Adventitious root formation is a key step in vegetative propagation and considerable progress has recently been made in understanding root formation. But, in spite of the efforts made, no new rooting treatments have been developed. Here, we report for the first time, that N,N'-bis-(2,3-methylenedioxyphenyl)urea and N,N'-bis-(3,4-methylenedioxyphenyl)urea enhance adventitious root formation in microcuttings of Malus pumila Mill. rootstock M26. Roots emerge without auxin supplementation in the darkness, transfer in hormone free medium, or callus formation. With the use of different bioassays, we also demonstrate that these two diphenylurea derivatives do not show cytokinin- or auxin-like activity.

Journal Article↗
PubMed2005

QTLs controlling the production of transgenic and adventitious roots in Brassica oleracea following treatment with Agrobacterium rhizogenes.

Brassica oleracea can be genetically engineered using Agrobacterium rhizogenes. The initial stage of this process is the production of transgenic ('hairy') roots; shoots are subsequently regenerated from these roots. Previous work using gus and gfp reporter genes has shown that genotypes of B. oleracea vary in their performance for transgenic root production. Quantitative trait loci (QTLs) controlling this trait have been located in one mapping population. The current study provides evidence that performance for transgenic root production is associated with performance for adventitious (non-transgenic) root production in B. oleracea across a second mapping population. This is shown by regression analyses between performance for the two traits and the demonstration that QTLs controlling the two traits map to the same positions within the genome. Since the rate of adventitious root production does not differ significantly in the presence and absence of A. rhizogenes, there is no evidence that the expression of Agrobacterium genes induces adventitious root production. It is apparent that genotypes exhibiting high adventitious root production in the absence of A. rhizogenes will also tend to show high transgenic root production, thereby allowing the selection of lines that are more efficiently transformed.

Brassica↗
PubMed2003

Genetic analysis of adventitious root formation with a novel series of temperature-sensitive mutants of Arabidopsis thaliana.

When cultured on media containing the plant growth regulator auxin, hypocotyl explants of Arabidopsis thaliana generate adventitious roots. As a first step to investigate the genetic basis of adventitious organogenesis in plants, we isolated nine temperature-sensitive mutants defective in various stages in the formation of adventitious roots: five root initiation defective (rid1 to rid5) mutants failed to initiate the formation of root primordia; in one root primordium defective (rpd1) mutant, the development of root primordia was arrested; three root growth defective (rgd1, rgd2, and rgd3) mutants were defective in root growth after the establishment of the root apical meristem. The temperature sensitivity of callus formation and lateral root formation revealed further distinctions between the isolated mutants. The rid1 mutant was specifically defective in the reinitiation of cell proliferation from hypocotyl explants, while the rid2 mutant was also defective in the reinitiation of cell proliferation from root explants. These two mutants also exhibited abnormalities in the formation of the root apical meristem when lateral roots were induced at the restrictive temperature. The rgd1 and rgd2 mutants were deficient in root and callus growth, whereas the rgd3 mutation specifically affected root growth. The rid5 mutant required higher auxin concentrations for rooting at the restrictive temperature, implying a deficiency in auxin signaling. The rid5 phenotype was found to result from a mutation in the MOR1/GEM1 gene encoding a microtubule-associated protein. These findings about the rid5 mutant suggest a possible function of the microtubule system in auxin response.

Arabidopsis↗
PubMed2007

Influence of IBA and aphidicolin on DNA synthesis and adventitious root regeneration from Malus 'Jork 9' stem discs.

Adventitious root formation in Malus 'Jork 9' stem discs was studied through temporarily blocking DNA synthesis by application of aphidicolin (AD). Higher number of roots per disc (8.4) after 21 days of cultivation were formed after a 24-h pulse of 15 microM AD, compared to control without AD application (6.7), with significantly more roots (3.7) already appearing at day 7, compared to 1.5 roots on the control. The promotive effect of AD on rooting was lower at 5 microM, while a concentration of 30 microM was slightly inhibitory. Results show that DNA synthesis is effectively blocked by AD, and this blockage is overcome after AD withdrawal. The data indicate that AD treatment influences cell divisions, thereby, might synchronise root initiation. The effects of different treatments with and without AD were studied at the cellular level by visualising DNA replication through BrdU-labelling. BrdU labelling further revealed temporal changes in the competence of the explants to respond to applied IBA. Thus, it is shown that the proportion of replicating nuclei present during 28-32 h is significantly increased in the split IBA treatment (0-4 h and 28-32 h; treatment C3), compared with a single IBA application during 0-8 h (treatment C3.1).

Aphidicolin↗
PubMed1999

Adventitious root growth and cell-cycle induction in deepwater rice

Deepwater rice (Oryza sativa) is adapted to survive conditions of severe flooding over extended periods of time. During such periods adventitious roots develop to provide water, nutrients, and anchorage. In the present study the growth of adventitious roots was induced by treatment with ethylene but not auxin, cytokinin, or gibberellin. Root elongation was enhanced between 8 and 10 h after submergence. The population of cells in the S phase and expression of the S-phase-specific histone H3 gene increased within 4 to 6 h. Within 6 to 8 h the G2-phase population increased. Cell-cycle activation was accompanied by sequential induction of a cdc2-activating kinase homolog, R2, of two cdc2 genes, cdc2Os-1 and cdc2Os-2, and of three cyclin genes, cycA1;3, cycB2;1, and cycB2;2, but only induction of the R2 gene expression preceded the induction of the S phase, possibly contributing to cell-cycle regulation in the G1 phase. Both cdc2 genes were expressed at slightly higher levels during DNA replication. Transcripts of the A-type cyclin accumulated during the S and G2 phases, and transcripts of the B-type cyclins accumulated during the G2 phase. Cyclin expression was induced at all nodes with a similar time course, suggesting that ethylene acts systemically and that root primordia respond to ethylene at an early developmental stage.

Journal Article↗
PubMed1991

Effect of Exogenous Indole-3-Acetic Acid and Indole-3-Butyric Acid on Internal Levels of the Respective Auxins and Their Conjugation with Aspartic Acid during Adventitious Root Formation in Pea Cuttings.

The influence of exogenous indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) on the internal levels of these auxins was studied during the first 4 days of adventitious root formation in cuttings of Pisum sativum L. The quantitations were done by high performance liquid chromatography with spectrofluorometric detection. IBA, identified by combined gas chromatography-mass spectrometry (GC-MS), was found to naturally occur in this plant material. The root inducing ability of exogenous IBA was superior to that of IAA. The IAA level in the tissue increased considerably on the first day after application of IAA, but rapidly decreased again, returning to a level twice the control by day 3. The predominant metabolic route was conjugation with aspartic acid, as reflected by the increase in the level of indole-3-acetylaspartic acid. The IBA treatment resulted in increases in the levels of IBA, IAA, and indole-3-acetylaspartic acid. The IAA content rapidly returned to control levels, whereas the IBA level remained high throughout the experimental period. High amounts of indole-3-butyrylaspartic acid were found in the tissue after feeding with IBA. The identity of the conjugate was confirmed by (1)H-nuclear magnetic resonance and GC-MS. IBA was much more stable in solution than IAA. No IAA was detected after 48 hours, whereas 70% IBA was still recovered after this time. The relatively higher root inducing ability of IBA is ascribed to the fact that its level remained elevated longer than that of IAA, even though IBA was metabolized in the tissue. Adventitious root formation is discussed on the basis of these findings.

Journal Article↗
PubMed2004

Nitrogen- and storage-affected carbohydrate partitioning in high-light-adapted Pelargonium cuttings in relation to survival and adventitious root formation under low light.

BACKGROUND AND AIMS: The aim of this study was to determine the role of nitrogen- and storage-affected carbohydrate availability in rooting of pelargonium cuttings, focusing on the environmental conditions of stock plant cultivation at low latitudes, transport of cuttings, and rooting under the low light that prevails during the winter rooting period in Central European greenhouses. METHODS: Carbohydrate partitioning in high-light-adapted cuttings of the cultivar 'Isabell' was studied in relation to survival and adventitious root formation under low light. Effects of a graduated supply of mineral nitrogen to stock plants and of cutting storage were examined. KEY RESULTS: Nitrogen deficiency raised starch levels in excised cuttings, whereas the concentrations of glucose and total sugars in leaves and the basal stem were positively correlated with internal total nitrogen (Nt). Storage reduced starch to trace levels in all leaves, but sugar levels were only reduced in tissues of non-nitrogen deficient cuttings. Sugars accumulated in the leaf lamina of stored cuttings during the rooting period, whereas carbohydrates were simultaneously exhausted in all other cutting parts including the petioles, thereby promoting leaf senescence. The positive correlation between initial Nt and root number disappeared after storage. Irrespectively of storage, higher pre-rooting leaf glucose promoted subsequent sugar accumulation in the basal stem and final root number. The positive relationships between initial sugar levels in the stems with cutting survival and in leaves with root formation under low light were confirmed in a sample survey with 21 cultivars provided from different sources at low latitudes. CONCLUSIONS: The results indicate that adventitious rooting of pelargonium cuttings can be limited by the initial amount of nitrogen reserves. However, this relationship reveals only small plasticity and is superimposed by a predominant effect of carbohydrate availability that depends on the initial leaf sugar levels, when high-light adaptation and low current light conditions impair net carbon assimilation.

Adaptation, Physiological↗
PubMed2005

Mineral nutrition and adventitious rooting in microcuttings of Eucalyptus globulus.

We characterized the adventitious rooting response of Eucalyptus globulus Labill. to various concentrations of calcium, nitrogen, phosphorus, iron, manganese, zinc, boron and copper. The parameters analyzed were percent rooting, root number, root length and mean rooting time. Root number and root length were significantly affected by mineral nutrition, whereas mean rooting time and rooting percentage seemed to be closely related to auxin availability. Root number was affected by calcium, nitrogen source and zinc, whereas root length was influenced by concentrations of phosphorus, iron and manganese, and by nitrogen source. Based on these results, we evaluated various combinations of several concentrations of these minerals in each rooting phase. Cuttings that were rooted in an optimized mineral nutrient medium and acclimatized to ex-vitro conditions for two months showed significantly higher survival after transplanting and drought stress than cuttings rooted in basal medium and treated in the same way.

Culture Media↗
PubMed2004

Differences in structure of adventitious roots in Salix clones with contrasting characteristics of cadmium accumulation and sensitivity.

Various clones of Salix spp. have contrasting characteristics of accumulation, translocation to shoots and of sensitivity to cadmium (Cd). The aim was to investigate the structure of adventitious roots and find out if differences between groups of clones in root anatomy accounted for differences in relation to Cd. Stem cuttings of eight clones of Salix spp. with different combinations of high or low Cd accumulation, translocation of Cd to shoots and sensitivity to Cd, were cultivated for 3 weeks in hydroponics containing 100 micro M Ca(NO(3))(2). No Cd was added in this experiment. Equal-sized roots were selected for hand-sectioning and fluorescence staining to detect the beginning of Casparian band formation and suberin lamellae deposition in endodermis. In addition, root apices were fixed, embedded, sectioned longitudinally and transversally, and stained. The image analysis system LUCIA was used for quantitative evaluation of tissue differences. The structure of adventitious root apices showed an intermediate-open type of root apical meristem and the clones differed in organization of root apices. Clones with low accumulation of Cd and high Cd tolerance had smaller meristematic zones and more extensive vacuolation of cells in the root apices than clones characterized by high accumulation of, and high sensitivity to, Cd. The apoplastic barriers, exo- and endodermis, were developed relatively close to the apex. In both layers the first ontogenetic stage, the Casparian band development, was followed by the second stage, the suberin lamellae deposition. This process started in the endodermis, preferentially against phloem poles, which is a common phenomenon also in other plant species. However, preferential development of exodermis in the sectors against phloem poles was observed in this study for the first time in plants. Development of endodermal Casparian bands in clones characterized by high accumulation of Cd occurred more distant from the root tip than in clones with low accumulation. Furthermore, the suberin lamellae were more distant from the root tip in clones with high translocation of Cd compared with those with low translocation. This indicates that apoplastic movement of Cd into the stele and the upward translocation may vary due to the endodermal anatomy. The proportion of root apoplastic barriers, exodermis and endodermis as well as epidermis to other root tissues was significantly increased in clones with higher tolerance to Cd ions, indicating the importance of these tissues in protection of the root against toxic effects of Cd.

Journal Article↗
PubMed2000

Different rRNA gene expression in primary and adventitious roots of Allium cepa L.

Sequentially used silver staining and in situ hybridization allowed to estimate the number of rDNA loci and their activity in meristematic cells of Allium cepa roots. In primary roots, obtained from germinated seeds, the rDNA probe hybridized with four chromosomes and showed four strong sites of hybridization. All of them displayed very clear positive silver staining. In cells of adventitious roots, from bulbs, only one pair of rRNA gene loci was active and after in situ hybridization showed strong signals while two other sites were very weak. The results indicate different transcriptional rRNA gene activity in meristematic cells of roots of different developmental origin. The reduction of the number of active rRNA loci can be the result of DNA methylation but the reduction mechanism of in situ hybridization sites in adventitious roots of Allium cepa remains an open question.

Allium↗
PubMed1989

Decreased Ethylene Biosynthesis, and Induction of Aerenchyma, by Nitrogen- or Phosphate-Starvation in Adventitious Roots of Zea mays L.

Plants of Zea mays L. cv TX5855 were grown in a complete, well oxygenated nutrient solution then subjected to nutrient starvation by omitting either nitrate and ammonium or phosphate from the solution. These treatments induced the formation of aerenchyma close to the apex of the adventitious roots that subsequently emerged from the base of the shoot, a response similar to that shown earlier to be induced by hypoxia. Compared with control plants supplied with all nutrients throughout, N- or P-starvation consistently depressed the rates of ethylene release by excised, 25 mm apical segments of adventitious roots. Some enzymes and substrates of the ethylene biosynthetic pathway were examined. The content of 1-amino cyclopropane-1-carboxylic acid (ACC) paralleled the differences in ethylene production rates, being depressed by N or P deficiency, while malonyl-ACC showed a similar trend. Activity of ACC synthase and of ethylene forming enzyme (g(-1) fresh weight) was also greater in control roots than in nutrient starved ones. These results indicate that much of the ethylene biosynthetic pathway is slowed under conditions of N- or P-starvation. Thus, by contrast to the effects of hypoxia, the induction of aerenchyma in roots of Zea mays by nutrient starvation is not related to an enhanced biosynthesis and/or accumulation of ethylene in the root tips.

Journal Article↗
PubMed2005

Analysis of indole-3-butyric acid-induced adventitious root formation on Arabidopsis stem segments.

Root induction by auxins is still not well understood at the molecular level. In this study a system has been devised which distinguishes between the two active auxins indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA). IBA, but not IAA, efficiently induced adventitious rooting in Arabidopsis stem segments at a concentration of 10 microM. In wild-type plants, roots formed exclusively out of calli at the basal end of the segments. Root formation was inhibited by 10 microM 3,4,5-triiodobenzoic acid (TIBA), an inhibitor of polar auxin transport. At intermediate IBA concentrations (3-10 microM), root induction was less efficient in trp1, a tryptophan auxotroph of Arabidopsis with a bushy phenotype but no demonstrable reduction in IAA levels. By contrast, two mutants of Arabidopsis with measurably higher levels of IAA (trp2, amt1) show root induction characteristics very similar to the wild type. Using differential display, transcripts specific to the rooting process were identified by devising a protocol that distinguished between callus production only and callus production followed by root initiation. One fragment was identical to the sequence of a putative regulatory subunit B of protein phosphatase 2A. It is suggested that adventitious rooting in Arabidopsis stem segments is due to an interaction between endogenous IAA and exogenous IBA. In stem explants, residual endogenous IAA is transported to the basal end of each segment, thereby inducing root formation. In stem segments in which the polar auxin transport is inhibited by TIBA, root formation does not occur.

Arabidopsis↗