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PubMed2000

[Effects of IAA and stimulated microgravity on formation of adventitious roots of Chinese cabbage].

Under the induction of indole-3-acetic acid (IAA), adventitious roots were differentiated on hypocotyl segments derived from seedlings of Chinese cabbage (Brassica campestris spp. pekinensis). IAA at concentration of 0.4-1.0 mg/L in solid MS medium incited many adventitious roots on hypocotyl segments. The earliest anatomic changes were observed on cut surface of hypocotyl segments under optical microscope 24 hours after IAA treatment: cytoplasmic and nuclear density became higher in a few of parenchytmatous cells adjacent to phloem in tissue of pericycle, followed by cell divisions. Lately, the dividing cells expanded and developed into root primordium from which root cap was differentiated. After five days, most roots protruded through hypocotyl cortex and appeared just below the cut surface. The rooting capacity of the segments derived from three regions of each hypocotyl was different. High level of IAA modified the polarity of root formation on segment inserted upside down and sucrose increased the function of IAA. Additionally, microgravity did not significantly change the rooting polarity under the condition of stimulated microgravity, but it increased the competence of explants to IAA treatment. The results presented here provided an experimental system for further investigation of molecular events associated with adventitious root initiation.

Brassica↗
PubMed2003

ILR2, a novel gene regulating IAA conjugate sensitivity and metal transport in Arabidopsis thaliana.

Plants can regulate levels of the auxin indole-3-acetic acid (IAA) by conjugation to amino acids or sugars, and subsequent hydrolysis of these conjugates to release active IAA. These less active auxin conjugates constitute the majority of IAA in plants. We isolated the Arabidopsis ilr2-1 mutant as a recessive IAA-leucine resistant mutant that retains wild-type sensitivity to free IAA. ilr2-1 is also defective in lateral root formation and primary root elongation. In addition, ilr2-1 is resistant to manganese- and cobalt-mediated inhibition of root elongation, and microsomal preparations from the ilr2-1 mutant exhibit enhanced ATP-dependent manganese transport. We used a map-based positional approach to clone the ILR2 gene, which encodes a novel protein with no predicted membrane-spanning domains that is polymorphic among Arabidopsis accessions. Our results demonstrate that ILR2 modulates a metal transporter, providing a novel link between auxin conjugate metabolism and metal homeostasis.

Amino Acid Sequence↗
PubMed2004

Isolation and characterization of an IAA-responsive gene from Gossypium barbadense L.

The full-length cDNA of an IAA-responsive gene was cloned from Gossypium barbadense L. (designated as Gbiaa-Re) by rapid amplification of cDNA ends (RACE). Gbiaa-Re gene was 1043-bp long and contained a 573-bp open reading frame encoding a polypeptide of 190 amino acid residues. Homology analysis revealed that Gbiaa-Re strongly resembled known plant IAA-responsive genes. The conserved integrated domain "AUX_IAA, AUX/IAA family" resided within the region from L11, to V190 of GbIAA-RE, and the 4 typically conserved domains of IAA-responsive gene family were all found in GbIAA-RE. The secondary structure of GbIAA-RE consisted of 20.53% alpha helix, 13.68% extended strand and 65.79% random coil. In total, 12 phosphorylation sites, 1 N-glycosylation site and 4 O-beta-GlcNAc attachment sites were predicted. Southern blot analysis indicated that Gbiaa-Re belonged to a low-copy gene family. Semi-quantitative PCR analysis indicated that the expression of Gbiaa-Re gene was inducible by IAA. Our studies suggested that Gbiaa-Re was a new member of plant AUX/IAA gene family.

Amino Acid Sequence↗
PubMed1968

Relationship Between Metabolism and the Lateral Transport of IAA in Corn Coleoptiles.

The lateral movement of IAA in coleoptiles of Zea mays has been investigated under aerobic and anaerobic conditions. The IAA-1-(14)C was supplied asymmetrically to the apical end of the segment. The results were as follows: A) In air more (14)C was found in the lower half of horizontal segments supplied with an upper donor than in the half opposite the donor in vertical segments. The enhanced lateral movement of (14)C in geotropically stimulated segments of corn coleoptiles under aerobic conditions has thus been confirmed. B) This increased lateral movement of (14)C in geotropically stimulated segments is greatly reduced, but is not completely abolished, under anaerobic conditions. C) The lateral movement of (14)C in vertical segments is significantly less under anaerobic conditions than in air. D) Under anaerobic conditions, the lateral movement of (14)C in horizontal segments can be reduced to the level found in vertical segments by pre-soaking the tissue in a 1 mm solution of the metabolic inhibitor sodium fluoride for 2 hours. The inhibitor has no effect on lateral movement of (14)C in vertical anaerobic segments. E) In air, sodium fluoride has no effect on the lateral movement of (14)C in either vertical or horizontal segments.On the assumption that the movement of (14)C reflects the movement of IAA, these results show that the enhanced polarized lateral movement of IAA observed in horizontal corn coleoptile segments is totally abolished only when both aerobic and anaerobic metabolism are arrested. In addition, there appears to be a small metabolically dependent component in the lateral movement of IAA in vertical segments supplied with an asymmetric donor in air.

Journal Article↗
PubMed1969

IAA Oxidase Inhibitors from Normal and Mutant Maize Plants.

Extracts of maize (Zea mays L.) plants contain substances which, in vitro, inhibit an indoleacetic acid (IAA) oxidase enzyme from maize. The extracts can be freed of inhibitors by dialysis or by passage through columns of polyvinylpyrrolidone powder. Inhibitor-free extracts contain an IAA oxidase enzyme which requires a phenolic co-factor and is stimulated by Mn(2+).IAA oxidase inhibitor and total phenol levels were compared for normal maize and for the maize mutant Knotted (Kn). In plants up to 18 days old the level of heat stable, water soluble IAA oxidase inhibitors increases with increasing dosage of the Kn allele. Increased inhibitor content is accompanied, but not paralleled, by increased content of total phenols. Although several inhibitors are present in crude extracts, most inhibition can be attributed to 1 compound. This compound is not destroyed by horseradish peroxidase in the absence of IAA. At pH 3.5 it is not extracted into ether, but it is rendered ether-extractable by incubation in 2 n KOH for 5 hr at room temperature. This compound is tentatively identified as an ester of ferulic acid and some unknown moiety.

Journal Article↗
PubMed1987

Rapid Effects of IAA on Cell Surface Proteins from Intact Carrot Suspension Culture Cells.

Suspension cultures of carrot (Daucus carrota L.) which had an absolute requirement for exogenously supplied auxin were grown in medium containing indoleacetic acid (IAA) as the sole auxin source. Putative cell surface proteins were extracted from the intact cells. Resupply of IAA to cultures partially depleted of auxin resulted in rapidly increased activities of three enzyme activities subsequently extracted. Two of the enzyme activities which increased, peroxidase and pectinesterase, have been implicated in the literature as important to cell wall development, structure, and growth. The other enzyme activity which was increased, IAA oxidase, may be involved in the degradation of IAA In vivo. Polypeptides in the extracts were found to increase equally as rapidly as the enzymes in response to IAA as determined with sodium dodecyl sulfate-polyacrylamide electrophoretic gels stained with silver. It is not known whether the changes in enzyme and polypeptide levels in the protein extracts were due to auxin effects on protein synthesis, transport, or extractability.

Journal Article↗
PubMed1987

Differences in epitope restriction of autoantibodies to native human insulin (IAA) and antibodies to heterologous insulin (IA).

The binding profiles of insulin autoantibodies (IAA) and insulin antibodies (IA) to highly purified species variants and fragments of insulin were studied in direct immunospecific enzyme-linked immunosorbent assay (ELISA) and indirect absorption experiments with insulins covalently coupled to Sepharose beads. Five of 10 IAA-containing sera from insulin-naive nondiabetics that bound whole human (H) insulin did not bind whole porcine (P) or whole bovine (B) insulins. These sera bound H insulin B-chain but not P B-chain or desalanated P insulin, suggesting they were dependent on the presence of threonine B30. The other 5 IAA-containing sera bound H, P, B, and desalanated porcine insulins, but only 1 bound isolated B-chains. All 10 IA-containing sera from insulin-treated diabetics bound H, P, B, and desalanated P insulins, but only 1 bound to human (and porcine) B-chain. Further binding studies with ovine, rabbit, rat, and guinea pig insulins confirmed the H (threonine B30) specificity of the 5 IAA-containing sera. B30 residues do not appear to be dominant, however, when insulin is administered exogenously. Instead, IA bind predominantly to A-chain or conformational determinants involving both chains. Scatchard analysis of a representative H insulin-specific IAA serum suggested that it contained a single binding affinity, whereas analysis of a representative IA-diabetic serum suggested it contained several different affinities.

Adult↗
PubMed1989

The time course of insulin autoantibodies (IAA) in the diabetes prone BB rat.

The BB rat is a widely used animal model for the study of insulin dependent diabetes. An enzyme linked immunosorbent assay, using purified rat insulin, was used to measure serial insulin autoantibodies (IAA) levels in coded sera from the BB/W/D rat colony in order to establish the time course of IAA. The animals included 26 diabetes-prone BB rats, six diabetes-resistant BB rats and six Wistar controls. There was an increase in both IAA frequency and titre with time in the diabetes-prone group: none were positive at 45 days, 17/19 (89%) were positive by day 90 and all were positive thereafter. Similar results were observed in the diabetes-resistant BB group (0/6 positive at day 51, 6/6 positive at day 90). None of the Wistar controls were positive at 105 days, although occasional positive sera were observed at 120 days. IAA seem to be acquired early on in the majority of BB rats, both diabetes-prone and diabetes-resistant, and much later, if at all, in controls. A clear homology of the MHC genes exists in both BB rat sublines, thus IAA appear to be a strain related phenomenon rather than a marker for IDDM.

Animals↗
PubMed1994

Characterization of high-frequency deletions in the iaa-containing plasmid, pIAA2, of Pseudomonas syringae pv. savastanoi.

The phytopathogenic bacterium Pseudomonas syringae pv. savastanoi causes olive and oleander knot disease. The bacterium induces the formation of tumorous galls by the synthesis and secretion of the plant hormones trans-zeatin riboside and indole-3-acetic acid into host intercellular spaces. An Italian oleander isolate, PB213, has been observed to lose the ability to synthesize IAA at high frequency, thus becoming non-pathogenic. The IAA genes, located on the 72-kb iaa-containing plasmid, pIAA2 were lost mainly due to two classes of deletions: 18 or 22 kb in length. Both classes of deletions had a common endpoint upstream of the IAA genes. The other endpoints were in areas that flanked the insertion sequence element IS51. The endpoints are in regions of repetitive DNA of at least 271 bp that have been designated a/b.

Base Sequence↗
PubMed1991

35S-beta-glucuronidase gene blocks biological effects of cotransferred iaa genes.

The iaaM and iaaH genes of Agrobacterium tumefaciens and Agrobacterium rhizogenes play an important role in crown gall and hairy root disease. The iaaM gene codes for tryptophan monooxygenase which converts tryptophan into indole-3-acetamide (IAM). IAM is converted into the auxin indole-3-acetic acid (IAA) by indoleacetamide hydrolase, encoded by the iaaH gene. In functional studies on the activity of the iaa genes of the TB region of the A. tumefaciens biotype III strain Tm4, the frequently used 35S-beta-glucuronidase (35S-UidA or GUS) marker gene was found to inhibit IAA synthesis and root induction encoded by the TB iaa genes. To exert this inhibition, the 35S-UidA gene must be cotransferred with the iaaH gene. The 35S promoter alone is sufficient to cause the inhibitory effect.

Amidohydrolases↗
PubMed1994

Islet cell antibodies (ICA), insulin autoantibodies (IAA), islet cell surface antibodies (ICSA) and C-peptide in 1031 school children in a population with a high background incidence of IDDM.

Islet cell antibodies (ICA), insulin autoantibodies (IAA) and islet cell surface antibodies (ICSA) together with C-peptide were determined in 1031 healthy schoolchildren to evaluate the frequency of autoimmune reactions towards endocrine pancreas and its relation to insulin secretion in non-diabetic children. The prevalence of ICA (levels > 6 JDF units) was 1.4% (14/1012) while 44 children (4.3%) were ICSA-positive and 40 (4%) had IAA. Girls had higher titres of ICSA than boys. Young children (7-8 years) more often had IAA than 12-13-year-old children who, however, had ICA three times more often than the young children. There were no clear associations between the different antibodies. Of the children, 2.4% had very low post-prandial serum C-peptide values (< or = 0.25 nmol/l). Serum C-peptide was higher in girls than in boys (P < 0.001) and in older children than in younger (P < 0.001). Girls with low levels of ICA had high C-peptide values, while girls with high ICA titers had low C-peptide values, the latter perhaps indicating partial beta cell loss. IAA and ICSA were not related to C-peptide values but both positive ICSA and high C-peptide values were most common in the autumn (P < 0.02 and P < 0.0001, respectively). One of the ICA-positive children developed diabetes in 1991, 4 years after the blood sample was taken. Since after 5 years only one of the children has developed IDDM, it can be concluded autoimmune reactions towards endocrine pancreas and insulin may occur in many children without the development of manifest diabetes. Those with high ICA titers may have lost so many beta cells that their insulin secretion is affected, which in some cases might lead to diabetes many years later.

Adolescent↗
PubMed2004

IAA production during germination of Orobanche spp. seeds.

Broomrapes (Orobanche spp.) are parasitic plants, whose growth and development fully depend on the nutritional connection established between the parasite and the roots of the respective host plant. Phytohormones are known to play a role in establishing the specific Orobanche-host plant interaction. The first step in the interaction is seed germination triggered by a germination stimulant secreted by the host-plant roots. We quantified indole-3-acetic acid (IAA) and abscisic acid (ABA) during the seed germination of tobacco broomrape (Orobanche ramosa) and sunflower broomrape (O. cumana). IAA was mainly released from Orobanche seeds in host-parasite interactions as compared to non-host-parasite interactions. Moreover, germinating seeds of O. ramosa released IAA as early as 24 h after the seeds were exposed to the germination stimulant, even before development of the germ tube. ABA levels remained unchanged during the germination of the parasites' seeds. The results presented here show that IAA production is probably part of a mechanism triggering germination upon the induction by the host factor, thus resulting in seed germination.

Abscisic Acid↗
PubMedDate not supplied

Cloning and characterisation of a basic IAA oxidase associated with root induction in Vitis vinifera.

Changes in apoplastic peroxidases during auxin-induced in vitro rooting of cultured grapevine (Vitis vinifera L. cv. Touriga) stems have been studied. The largest increase in peroxidase activity (EC 1.11.1.7) was associated with the early stages of root initiation and could be attributed to an increase in activity of an apoplastic 36 kDa cationic peroxidase (PxB2). Relative to other peroxidases, PxB2 demonstrated high indole-3-acetic acid (IAA) oxidase activity and apparently contributed the majority of potential IAA oxidase activity in rooting tissues. The distribution of this peroxidase in developing roots additionally associates it with early phases of growth restriction. PxB2 was purified from cell wall extracts prepared from the basal 1 cm of rooting stems. Microsequencing and subsequent cloning of its corresponding 3' truncated cDNA (encoding 255 amino acids of the mature protein) revealed it to have a typical class III peroxidase structure. The results suggest that this class III peroxidase with IAA oxidase activity is important for the control of IAA levels during root initiation and development.

Amino Acid Sequence↗
PubMed1984

Rapid reduction by IAA of malondialdehyde levels in avena coleoptiles, a possible effect on lipid peroxidation.

Treatment of Avena coleoptile sections with IAA results in a rapid decrease in the level of lipid peroxidation (LP), as measured by the thiobarbituric acid reaction for malondialdehyde (MDA). The response is specific for active auxins, is nearly saturated by 10(-6) M IAA and occurs even when turgor is reduced by 0.1 M mannitol. About half the reduction in MDA occurs within 1 min after addition of IAA to intact tissues; addition of IAA directly to homogenates, however, has no effect. Homogenates prepared from auxin-pretreated tissues, but not control tissues, continue to produce MDA over at least a six-hour period. This effect of auxin on LP is one of the most rapid biochemical responses to auxin known, and suggests that LP might alter the properties of membranes and thus influence cell enlargement.

Indoleacetic Acids↗
PubMed2003

Effect of BAP and IAA on the expression of G1 and G2 control points and G1-S and G2-M transitions in root meristem cells of Vicia faba.

Excised, carbohydrate-starved root meristems of Vicia faba subsp. minor have been used to investigate the impact of the auxin indole-3-acetic acid (IAA) and the cytokinin benzyl-6-aminopurine (BAP) on (1) the expression of Principal Control Points (PCPs) during the G1- and G2-phases of the cell cycle, and (2) the dynamics of sucrose-mediated resumption of DNA replication and mitosis (G1-to-S and G2-to-M transitions). Compared with the excised root tips starved in mineral medium without hormones, stationary phase meristems induced during continuous treatment with BAP, IAA, or a mixture of BAP+IAA, increased the number of G2 cells, producing characteristic profiles of nuclear DNA content. In medium containing 2% sucrose, BAP accelerated PCP1-->S and PCP2-->M, whereas continuous treatment with IAA resulted in marked prolongation of both transitions. Since the PCPs regulate progression through the key events of interphase and mitosis by interacting with cyclin dependent kinases (CDKs), these results seem to correspond with current data indicating functional connections between phytohormones, nutritional signals, gene expression and the cell division cycles in plants.

Benzyl Compounds↗
PubMed1999

Redundancy as a way of life - IAA metabolism.

Plants have evolved elaborate systems for regulating cellular levels of indole-3-acetic acid (IAA). The redundancy of this network has complicated the elucidation of IAA metabolism, but molecular genetic studies and precise analytical methods have begun to expose the circuitry. It is now clear that plants synthesize, inactivate and catabolize IAA by multiple pathways, and multiple genes can encode a particular enzyme within a pathway. A number of these genes are now cloned, which greatly facilitates the future dissection of IAA metabolism.

Aminohydrolases↗
PubMed2004

Inorganic salts influence IAA ionization and adventitious rhizogenesis in Pongamia pinnata.

The basal cut end of coppice shoot cuttings of Pongamia pinnata was treated for 24 h with 0 (water treated control) or 5.0 mmol/L of KMnO4, KCI, and KH2PO4 or 2.5 mmol/L of K2HPO4 and K2SO4. Inorganic salts of P, S, Cl and Mn significantly influenced IAA ionization and adventitious rhizogenesis. P and S salts had lower IAA ionization potential, but more pronounced effect on adventitious rhizogenesis than Cl and Mn salts. The linear regression analysis also established negative correlations between salt induced IAA ionization with various characteristics of adventitious rhizogenesis such as sprouting (r = -0.83, p < 0.05), rooting (r = -0.82, p < 0.05), root number (r = -0.95, p < 0.01), and root length (r = -0.80, p < 0.1). The implication of IAA ionization in adventitious rhizogenesis has been discussed and the possible role of inorganic salts therein suggested.

Ions↗
PubMed2002

Resistance of transgenic tobacco seedlings expressing the Agrobacterium tumefaciens C58-6b gene, to growth-inhibitory levels of cytokinin is associated with elevated IAA levels and activation of phenylpropanoid metabolism.

We previously reported that the Agrobacterium tumefaciens C58-6b gene confers resistance to growth-inhibitory levels of exogenously applied N(6)-benzyladenine (BA, cytokinin) in transgenic tobacco (Nicotiana tabacum) seedlings. Here, we found that intracellular levels of indoleacetic acid (IAA, auxin) increased in transgenics but declined in wild-type seedlings upon BA treatment. Since exogenously supplied 1-naphthalene acetic acid (NAA), a stable synthetic auxin, counteracted the growth inhibition of wild-type seedlings by BA, we suggest that BA-induced growth inhibition in wild-type seedlings occurs, at least in part, as a result of intracellular IAA deficiency. Further HPLC analysis of cell extracts from BA-treated seedlings revealed that a fluorescent compound, later identified as the phenylpropanoid, scopolin, and the major phenolic compound, chlorogenic acid, accumulated earlier in transgenics than in wild-type seedlings. Gene transcripts encoding phenylalanine ammonia-lyase, cinnamate 4-hydroxylase, and 4-coumarate:CoA ligase, which are responsible for the early steps of phenylpropanoid biosynthesis, accumulated earlier and to higher levels in transgenics than in wild-type seedlings as determined by Northern hybridization analysis, thus accounting for the early accumulation of scopolin and chlorogenic acid in transgenics. As some phenolic compounds, including chlorogenic acid and scopoletin (aglycon of scopolin) are suggested to inhibit IAA catabolism, we further propose that C58-6b gene expression protects IAA from degradation by inducing the early phenylpropanoid pathway.

Adenine↗