Auxin production by plant associated bacteria: impact on endogenous IAA content and growth of Triticum aestivum L.

AIMS: The aim of this study was to investigate the potential of bacterial strains of Bacillus, Pseudomonas, Escherichia, Micrococcus and Staphylococcus genera associated with wild herbaceous flora to enhance endogenous indole-3-acetic acid (IAA) content and growth of Triticum aestivum var. Inqalab-91. METHODS AND RESULTS: Gas chromatography and mass spectrometric (GC-MS) analysis revealed that bacterial strains produced 0.6-8.22 microg IAA ml(-1) in the presence of L-tryptophan. Plant microbe experiments showed a significant positive correlation between auxin production by bacterial strains and endogenous IAA content of T. aestivum for GC-MS (r = 0.618; P = 0.05) and colorimetric analysis (r = 0.693; P = 0.01). Similarly, highly significant positive correlation for shoot length (r = 0.627; P = 0.01) and shoot fresh weight (r = 0.626; P = 0.01) was observed with auxin production under axenic conditions. Bacterial inoculations also enhanced shoot length (up to 29.16%), number of tillers (up to 97.35%), spike length (up to 25.20%) and seed weight (up to 13.70%) at final harvest. CONCLUSIONS: Bacterial strains have the ability to increase the endogenous IAA content and growth of T. aestivum var. Inqalab-91. SIGNIFICANCE AND IMPACT OF THE STUDY: Microbial strains of wild herbaceous flora can be effectively used to enhance the growth and yield of agronomically important crops.

Mercury in urban soils: a comparison of local spatial variability in six European cities.

The objective of this study was to quantify and assess for the first time the variability of total mercury in urban soils at a European level, using a systematic sampling strategy and a common methodology. We report results from a comparison between soil samples from Aveiro (Portugal), Glasgow (Scotland), Ljubljana (Slovenia), Sevilla (Spain), Torino (Italy) and Uppsala (Sweden). At least 25 sampling points (in about 4-5 ha) from a park in each city were sampled at two depths (0-10 and 10-20 cm). Total mercury was determined by pyrolysis atomic absorption spectrometry with gold amalgamation. The quality of results was monitored using certified reference materials (BCR 142R and BCR 141R). Measured total mercury contents varied from 0.015 to 6.3 mg kg(-1). The lowest median values were found in Aveiro, for both surface (0-10 cm) and sub-surface (10-20 cm) samples (0.055 and 0.054 mg kg(-1), respectively). The highest median mercury contents in soil samples were found in samples from Glasgow (1.2 and 1.3 mg kg(-1), for surface and sub-surface samples, respectively). High variability of mercury concentrations was observed, both within each park and between cities. This variability reflecting contributions from natural background, previous anthropogenic activities and differences in the ages of cities and land use, local environmental conditions as well as the influence of their location within the urban area. Short-range variability of mercury concentrations was found to be up to an order of magnitude over the distance of only a few 10 m.

Effect of light on growth and endogenous hormones in Chlorella minutissima (Trebouxiophyceae).

Plant growth regulators (PGRs) play an important role in mediating growth and stress responses in plants. Light influences PGRs concentrations in vascular plants. The effect of light on growth and endogenous PGR concentrations in microalgae was investigated in the present study. Chlorella minutissima MACC 360 was grown in 14:10 h light:dark (L:D), continuous dark (CD) and continuous dark with the addition of 5 g L(-1) glucose (CD + G) for 48 h. Cultures were synchronized in the L:D cultures, increasing in size during the light period and dividing during the dark period. C. minutissima cells did not increase in size or undergo cell division in CD cultures. In CD + G conditions, the cultures were no longer synchronized but did continue to increase in cell size and constantly underwent cell division although fewer cells divided than in the L:D cultures. Endogenous auxin and cytokinin concentrations increased and gibberellin concentrations decreased over time in the actively growing cultures (L:D and CD + G) but did not increase in the CD cultures. The largest increase in indole content was in the CD + G cultures while the L:D cultures had the largest cytokinin increase. Brassinosteroid concentrations decreased over time in all the cultures including those grown in CD conditions. Abscisic acid (ABA) concentrations were low and only increased in the CD cultures. These results show that endogenous PGRs were affected by the light regime and/or culture growth.

Natural and anthropogenic metal inputs to soils in urban Uppsala, Sweden.

Urban soils are complex systems due to human activities that disturb the natural development of the soil horizons and add hazardous elements. Remediation projects are common in urban areas and guideline values are set to represent a desired level of elements. However, the natural content of trace elements may not always equal the desired levels. In this study, an attempt is made to distinguish between metals that are present in the soil due to natural origins and to anthropogenic origins. Seventy-five soil samples of the 0-5, 5-10 and 10-20 cm layers were collected from 25 sites in urban areas of Uppsala City and analysed for aluminium (Al), arsenic (As), cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), mercury (Hg), manganese (Mn), nickel (Ni), lead (Pb), tungsten (W) and zinc (Zn) using aqua regia for digestion. In order to highlight elements of geological origin, the results were compared to a similar study carried out in Gothenburg City, which has about three times as many inhabitants as Uppsala and has a more industrial history. A cluster analysis was also performed to distinguish between elements of natural and anthropogenic origin. Contents of As, Al, Fe, Cr, Ni, Mn and W in Uppsala were concluded to be of mainly geological origin, while contents of Cd, Cu, Zn, Pb and Hg seemed to have been impacted upon by mainly urban activities.

Variability in concentrations of potentially toxic elements in urban parks from six European cities.

Use of a harmonised sampling regime has allowed comparison of concentrations of copper, chromium, nickel, lead and zinc in six urban parks located in different European cities differing markedly in their climate and industrial history. Wide concentrations ranges were found for copper, lead and zinc at most sites, but for chromium and nickel a wide range was only seen in the Italian park, where levels were also considerably greater than in other soils. As might be expected, the soils from older cities with a legacy of heavy manufacturing industry (Glasgow, Torino) were richest in potentially toxic elements (PTEs); soils from Ljubljana, Sevilla and Uppsala had intermediate metal contents, and soils from the most recently established park, in the least industrialised city (Aveiro), displayed lowest concentrations. When principal component analysis was applied to the data, associations were revealed between pH and organic carbon content; and between all five PTEs. When pH and organic carbon content were excluded from the PCA, a distinction became clear between copper, lead and zinc (the "urban" metals) on the one hand, and chromium and nickel on the other. Similar results were obtained for the surface (0-10 cm depth) and sub-surface (10-20 cm depth) samples. Comparisons with target or limit concentrations were limited by the existence of different legislation in different countries and the fact that few guidelines deal specifically with public-access urban soils intended for recreational use.

Sites and homeostatic control of auxin biosynthesis in Arabidopsis during vegetative growth.

The distribution and biosynthesis of indole-3-acetic acid (IAA) was investigated during early plant development in Arabidopsis. The youngest leaves analysed, less than 0.5 mm in length, contained 250 pg mg(-1) of IAA and also exhibited the highest relative capacity to synthesize this hormone. A decrease of nearly one hundred-fold in IAA content occurred as the young leaves expanded to their full size, and this was accompanied by a clear shift in both pool size and IAA synthesis capacity. The correlation between high IAA content and intense cell division was further verified in tobacco leaves, where a detailed analysis revealed that dividing mesophyll tissue contained ten-fold higher IAA levels than tissue growing solely by elongation. We demonstrated that all parts of the young Arabidopsis plant can potentially contribute to the auxin needed for growth and development, as not only young leaves, but also all other parts of the plant such as cotyledons, expanding leaves and root tissues have the capacity to synthesize IAA de novo. We also observed that naphthylphthalamic acid (NPA) treatment induced tissue-dependent feedback inhibition of IAA biosynthesis in expanding leaves and cotyledons, but intriguingly not in young leaves or in the root system. This observation supports the hypothesis that there is a sophisticated tissue-specific regulatory mechanism for auxin biosynthesis. Finally, a strict requirement for maintaining the pool sizes of IAA was revealed as reductions in leaf expansion followed both decreases and increases in the IAA levels in developing leaves. This indicates that leaves are not only important sources for IAA synthesis, but that normal leaf expansion depends on rigorous control of IAA homeostasis.

Developmental regulation of indole-3-acetic acid turnover in Scots pine seedlings.

Indole-3-acetic acid (IAA) homeostasis was investigated during seed germination and early seedling growth in Scots pine (Pinus sylvestris). IAA-ester conjugates were initially hydrolyzed in the seed to yield a peak of free IAA prior to initiation of root elongation. Developmental regulation of IAA synthesis was observed, with tryptophan-dependent synthesis being initiated around 4 d and tryptophan-independent synthesis occurring around 7 d after imbibition. Induction of catabolism to yield 2-oxindole-3-acetic acid and irreversible conjugation to indole-3-acetyl-N-aspartic acid was noticed at the same time as de novo synthesis was first detected. As a part of the homeostatic regulation IAA was further metabolized to two new conjugates: glucopyranosyl-1-N-indole-3-acetyl-N-aspartic acid and glucopyranosyl-1-N-indole-3-acetic acid. The initial supply of IAA thus originates from stored pools of IAA-ester conjugates, mainly localized in the embryo itself rather than in the general nutrient storage tissue, the megagametophyte. We have found that de novo synthesis is first induced when the stored pool of conjugated IAA is used up and additional hormone is needed for elongation growth. It is interesting that when de novo synthesis is induced, a distinct induction of catabolic events occurs, indicating that the seedling needs mechanisms to balance synthesis rates for the homeostatic regulation of the IAA pool.

Blue light regulates an auxin-induced K+-channel gene in the maize coleoptile.

Auxin redistribution along gravistimulated maize coleoptiles causes differential expression of the auxin-induced K+-channel gene ZMK1 (Zea mays K+ channel 1) and precedes the curvature response. To evaluate the role of ZMK1 during phototropism, we here investigated blue light-stimulated coleoptiles. Four hours of blue light stimulation resulted in phototropic bending (23 degrees ). Rotation on a clinostat, at nominally "zero" gravity, and simultaneous stimulation with unidirectional blue light, however, resulted in up to 51 degrees bending toward the light. Differential ZMK1 transcription reached a maximum after 90 min of blue light stimulation under gravity, whereas ZMK1 expression remained asymmetric for at least 180 min in photostimulated coleoptiles on a clinostat. We therefore conclude that the stronger phototropic bending under nominally "zero" gravity results from prolonged differential expression of ZMK1. Under both conditions, asymmetric expression of ZMK1 could be superimposed on the lateral auxin gradient across the coleoptile tip, whereas the gene for the blue light receptor phototropin 1 (PHOT1), expressed in the tip only, was not differentially regulated in response to blue light. The activation of the two different receptors eliciting the photo- and gravitropic response of the coleoptile thus feeds into a common signaling pathway, resulting in auxin redistribution in the coleoptile tip and finally in differential transcription of ZMK1. In the process of signal integration, gravity transduction restricts the magnitude of the blue light-inducible ZMK1 gradient. The spatial and temporal distribution of ZMK1 transcripts and thus differential K+ uptake in both flanks of the coleoptile seem to limit the stimulus-induced bending of this sensory organ.

Localization of the auxin permease AUX1 suggests two functionally distinct hormone transport pathways operate in the Arabidopsis root apex.

Auxins represent an important class of plant hormone that regulate plant development. Plants use specialized carrier proteins to transport the auxin indole-3-acetic acid (IAA) to target tissues. To date, efflux carrier-mediated polar auxin transport has been assumed to represent the sole mode of long distance IAA movement. Localization of the auxin permease AUX1 in the Arabidopsis root apex has revealed a novel phloem-based IAA transport pathway. AUX1, asymmetrically localized to the plasma membrane of root protophloem cells, is proposed to promote the acropetal, post-phloem movement of auxin to the root apex. MS analysis shows that IAA accumulation in aux1 mutant root apices is impaired, consistent with an AUX1 phloem unloading function. AUX1 localization to columella and lateral root cap tissues of the Arabidopsis root apex reveals that the auxin permease regulates a second IAA transport pathway. Expression studies using an auxin-regulated reporter suggest that AUX1 is necessary for root gravitropism by facilitating basipetal auxin transport to distal elongation zone tissues.

Control of axillary bud initiation and shoot architecture in Arabidopsis through the SUPERSHOOT gene.

The aerial architecture of flowering plants is determined to a large extent by shoot growth and shoot branching arising from the initiation and growth of axillary meristems. We have identified an Arabidopsis mutant, supershoot (sps), which is characterized by a massive overproliferation of shoots, such that a single plant can generate 500 or more inflorescences. Analysis of the mutant plants shows that the primary defect is because of an increase in the number of meristems formed in leaf axils, together with release of bud arrest, resulting in reiterative branch formation from rosette and cauline leaves. The SPS gene is shown here to encode a cytochrome P450, and together with a 3- to 9-fold increase in levels of Z-type cytokinins in sps mutant plants, indicate a role for SPS in modulating hormone levels. The expression pattern of SPS, with strong expression at the leaf axils, correlates well with the phenotypic defects. Our results indicate that control of shoot branching in Arabidopsis may be accomplished in part by suppression of axillary meristem initiation and growth through the localized attenuation of cytokinin levels at sites of bud initiation.

The SUR2 gene of Arabidopsis thaliana encodes the cytochrome P450 CYP83B1, a modulator of auxin homeostasis.

Genetic screens have been performed to identify mutants with altered auxin homeostasis in Arabidopsis. A tagged allele of the auxin-overproducing mutant sur2 was identified within a transposon mutagenized population. The SUR2 gene was cloned and shown to encode the CYP83B1 protein, which belongs to the large family of the P450-dependent monooxygenases. SUR2 expression is up-regulated in sur1 mutants and induced by exogenous auxin in the wild type. Analysis of indole-3-acetic acid (IAA) synthesis and metabolism in sur2 plants indicates that the mutation causes a conditional increase in the pool size of IAA through up-regulation of IAA synthesis.

Auxin-induced K+ channel expression represents an essential step in coleoptile growth and gravitropism.

Auxin-induced growth of coleoptiles depends on the presence of potassium and is suppressed by K+ channel blockers. To evaluate the role of K+ channels in auxin-mediated growth, we isolated and functionally expressed ZMK1 and ZMK2 (Zea mays K+ channel 1 and 2), two potassium channels from maize coleoptiles. In growth experiments, the time course of auxin-induced expression of ZMK1 coincided with the kinetics of coleoptile elongation. Upon gravistimulation of maize seedlings, ZMK1 expression followed the gravitropic-induced auxin redistribution. K+ channel expression increased even before a bending of the coleoptile was observed. The transcript level of ZMK2, expressed in vascular tissue, was not affected by auxin. In patch-clamp studies on coleoptile protoplasts, auxin increased K+ channel density while leaving channel properties unaffected. Thus, we conclude that coleoptile growth depends on the transcriptional up-regulation of ZMK1, an inwardly rectifying K+ channel expressed in the nonvascular tissue of this organ.