The development of yellow lupin anthers depends on the relationship between jasmonic acid and indole-3-acetic acid.

The main purpose of this study was to demonstrate that the course of anther development, including post-meiotic maturation, dehiscence and senescence, is ensured by the interdependencies between jasmonic acid (JA) and indole-3-acetic acid (IAA) in yellow lupin (Lupinus luteus L.). The concentration of JA peaked during anther dehiscence when IAA level was low, whereas the inverse relationship was specific to anther senescence. Cellular and tissue localization of JA and IAA, in conjunction with broad expression profile for genes involved in biosynthesis, signalling, response, and homeostasis under different conditions, allowed to complete and define the role of studied phytohormones during late anther development, as well as predict events triggered by them. The development/degeneration of septum and anther wall cells, dehydration of epidermis, and rupture of stomium may involve JA signalling, while the formation of secondary thickening in endothecial cell walls is rather JA independent. The IAA is involved in programmed cell death (PCD)-associated processes during anther senescence but does not exclude its participation in the anther dehiscence processes, mainly related to cell disintegration and degeneration. A detailed understanding of these multistage processes, especially at the level of phytohormonal interplay, can contribute to the effective control of male fertility, potentially revolutionizing the breeding of L. luteus.

Anther dehiscence is regulated by gibberellic acid in yellow lupine (Lupinus luteus L.).

BACKGROUND: Anther dehiscence resulting in the release of pollen grains is tightly regulated in a spatiotemporal manner by various factors. In yellow lupine (Lupinus luteus L.), a species that shows cleistogamy, the anthers split before the flowers open, but the course and regulation of this process are unknown. The specific control of anther development takes place via hormonal pathways, the wide action of which ensures reproductive success. In our previous research concerning flower and early pod development in yellow lupine, we showed that the lowest transcript level of LlDELLA1, a main repressor of gibberellin (GA) signalling, occurs approximately at the time of anther opening; therefore, the main purpose of this study was to precisely investigate the gibberellic acid (GA3)-dependent regulation of the anther dehiscence in this species. RESULTS: In this paper, we showed the specific changes in the yellow lupine anther structure during dehiscence, including secondary thickening in the endothecium by lignocellulosic deposition, enzymatic cell wall breakdown at the septum/stomium and cell degeneration via programmed cell death (PCD), and identified several genes widely associated with this process. The expression profile of genes varied over time, with the most intense mRNA accumulation in the phases prior to or at the time of anther opening. The transcriptional activity also revealed that these genes are highly coexpressed and regulated in a GA-dependent manner. The cellular and tissue localization of GA3 showed that these molecules are present before anther opening, mainly in septum cells, near the vascular bundle and in the endothecium, and that they are subsequently undetectable. GA3 localization strongly correlates with the transcriptional activity of genes related to GA biosynthesis and deactivation. The results also suggest that GA3 controls LlGAMYB expression via an LlMIR159-dependent pathway. CONCLUSIONS: The presented results show a clear contribution of GA3 in the control of the extensive anther dehiscence process in yellow lupine. Understanding the processes underlying pollen release at the hormonal and molecular levels is a significant aspect of controlling fertility in this economically important legume crop species and is of increasing interest to breeders.

EPIP-Evoked Modifications of Redox, Lipid, and Pectin Homeostasis in the Abscission Zone of Lupine Flowers.

Yellow lupine is a great model for abscission-related research given that excessive flower abortion reduces its yield. It has been previously shown that the EPIP peptide, a fragment of LlIDL (INFLORESCENCE DEFICIENT IN ABSCISSION) amino-acid sequence, is a sufficient molecule to induce flower abortion, however, the question remains: What are the exact changes evoked by this peptide locally in abscission zone (AZ) cells? Therefore, we used EPIP peptide to monitor specific modifications accompanied by early steps of flower abscission directly in the AZ. EPIP stimulates the downstream elements of the pathway-HAESA and MITOGEN-ACTIVATED PROTEIN KINASE6 and induces cellular symptoms indicating AZ activation. The EPIP treatment disrupts redox homeostasis, involving the accumulation of H2O2 and upregulation of the enzymatic antioxidant system including superoxide dismutase, catalase, and ascorbate peroxidase. A weakening of the cell wall structure in response to EPIP is reflected by pectin demethylation, while a changing pattern of fatty acids and acyl lipids composition suggests a modification of lipid metabolism. Notably, the formation of a signaling molecule-phosphatidic acid is induced locally in EPIP-treated AZ. Collectively, all these changes indicate the switching of several metabolic and signaling pathways directly in the AZ in response to EPIP, which inevitably leads to flower abscission.

Gibberellin Signaling Repressor LlDELLA1 Controls the Flower and Pod Development of Yellow Lupine (Lupinus luteus L.).

Precise control of generative organ development is of great importance for the productivity of crop plants, including legumes. Gibberellins (GAs) play a key role in the regulation of flowering, and fruit setting and development. The major repressors of GA signaling are DELLA proteins. In this paper, the full-length cDNA of LlDELLA1 gene in yellow lupine (Lupinus luteus L.) was identified. Nuclear-located LlDELLA1 was clustered in a second phylogenetic group. Further analyses revealed the presence of all conserved motifs and domains required for the GA-dependent interaction with Gibberellin Insensitive Dwarf1 (GID1) receptor, and involved in the repression function of LlDELLA1. Studies on expression profiles have shown that fluctuating LlDELLA1 transcript level favors proper flower and pod development. Accumulation of LlDELLA1 mRNA slightly decreases from the flower bud stage to anther opening (dehiscence), while there is rapid increase during pollination, fertilization, as well as pod setting and early development. LlDELLA1 expression is downregulated during late pod development. The linkage of LlDELLA1 activity with cellular and tissue localization of gibberellic acid (GA3) offers a broader insight into the functioning of the GA pathway, dependent on the organ and developmental stage. Our analyses provide information that may be valuable in improving the agronomic properties of yellow lupine.

The Role of Saccharides in the Mechanisms of Pathogenicity of Fusarium oxysporum f. sp. lupini in Yellow Lupine (Lupinus luteus L.).

The primary aim of this study was to determine the relationship between soluble sugar levels (sucrose, glucose, or fructose) in yellow lupine embryo axes and the pathogenicity of the hemibiotrophic fungus Fusarium oxysporum f. sp. Schlecht lupini. The first step of this study was to determine the effect of exogenous saccharides on the growth and sporulation of F. oxysporum. The second one focused on estimating the levels of ergosterol as a fungal growth indicator in infected embryo axes cultured in vitro on sugar containing-medium or without it. The third aim of this study was to record the levels of the mycotoxin moniliformin as the most characteristic secondary metabolite of F. oxysporum in the infected embryo axes with the high sugar medium and without it. Additionally, morphometric measurements, i.e., the length and fresh weight of embryo axes, were done. The levels of ergosterol were the highest in infected embryo axes with a sugar deficit. At the same time, significant accumulation of the mycotoxin moniliformin was recorded in those tissues. Furthermore, it was found that the presence of sugars in water agar medium inhibited the sporulation of the pathogenic fungus F. oxysporum in relation to the control (sporulation of the pathogen on medium without sugar), the strongest inhibiting effect was observed in the case of glucose. Infection caused by F. oxysporum significantly limited the growth of embryo axes, but this effect was more visible on infected axes cultured under sugar deficiency than on the ones cultured with soluble sugars. The obtained results thus showed that high sugar levels may lead to reduced production of mycotoxins by F. oxysporum, limiting infection development and fusariosis.

Melatonin Inhibits Peroxide Production in Plant Mitochondria.

The effect of melatonin on respiration and production (release) of hydrogen peroxide during succinate oxidation in mitochondria isolated from lupine cotyledons and epicotyls of pea seedlings was studied. It was shown for the first time that melatonin (10-7-10-3 M) had a significant inhibitory effect on the production of peroxide by plant mitochondria, which was characterized by concentration dependence and species specificity. At the same time, melatonin (at a concentration of up to 100 µM) had virtually no effect on mitochondrial respiration rate and respiratory control coefficient. The results confirm the antioxidant function of melatonin and indicate that it is involved in the regulation of ROS levels and maintenance of redox balance in plant mitochondria.

Effect of herbicides in the oxidative stress in crop winter species.

Most herbicides applied in crop field, stay in the soil for a period, affecting next crop or even the plants using as green manure. Nowadays, the use of herbicides grow to increase productivity, mainly in the grain producing region north of Rio Grande do Sul state. The objective of this study was to evaluate the effects of herbicides fomesafen and sulfentrazone on antioxidant system in Avena sativa1, Vicia sativa2, Raphanus sativus and Lupinus albus. The plants were exposed to varying concentrations of fomesafen3 (0, 0.125, 0.25 and 0.5 kg ha -1) and sulfentrazone (0, 0.3, 0.6 and 1.2 kg ha-1). For this, the activities of, ascorbat peroxidase, catalase and guaiacol enzymes were analyzed, and the levels of MDA were quantificated. Fomesafen and sulfentrazone promoted alterations in balance of plants generating oxidative stress and elicited the response of the antioxidant system of plants, mainly in the high doses of fomesafen, for the species V. sativa and R. sativus. At the same time, the 1,2 kg ha -1 dose of sulfentrazone generated lipid peroxidation for V. sativa, R. sativus and L. albus. Additionally, A. sativa was the species that demonstrated low alterations on antioxidant system with the exposure to herbicide fomesafen and sulfentrazone. Thus, we can we can suggest that the species present a better response in defense of the oxidative stress generated by the herbicides.

Long-Term Effects of Cold on Growth, Development and Yield of Narrow-Leaf Lupine May Be Alleviated by Seed Hydropriming or Butenolide.

In this article, the effects of cold on the development of Lupine angustifolius and the possibility of mitigating it, via seed hydropriming or pre-treatment with butenolide (10-6 M⁻10-4 M), are investigated in two cultivars, differing in their ability to germinate at low temperature. Physiological background of plant development after cold stress was investigated in imbibed seeds. For the first four weeks, the seedlings grew at 7 °C or 13 °C. Seeds well germinating at 7 °C demonstrated higher activity of α-amylase and higher levels of gibberellins, IAA and kinetin. Germination ability at low temperature correlated with dehydrogenase activity and membrane permeability. Seed pre-treatment improved germination at low temperature by decreasing abscisic acid content. Seed hydropriming alleviated cold effects on plant development rate and yield, while butenolide accelerated vegetative development but delayed the generative phase. Potential seed yield may be predicted based on the seed germination vigour and the photosynthetic efficiency measured before flowering.

Immunolocalization of dually phosphorylated MAPKs in dividing root meristem cells of Vicia faba, Pisum sativum, Lupinus luteus and Lycopersicon esculentum.

KEY MESSAGE: In plants, phosphorylated MAPKs display constitutive nuclear localization; however, not all studied plant species show co-localization of activated MAPKs to mitotic microtubules. The mitogen-activated protein kinase (MAPK) signaling pathway is involved not only in the cellular response to biotic and abiotic stress but also in the regulation of cell cycle and plant development. The role of MAPKs in the formation of a mitotic spindle has been widely studied and the MAPK signaling pathway was found to be indispensable for the unperturbed course of cell division. Here we show cellular localization of activated MAPKs (dually phosphorylated at their TXY motifs) in both interphase and mitotic root meristem cells of Lupinus luteus, Pisum sativum, Vicia faba (Fabaceae) and Lycopersicon esculentum (Solanaceae). Nuclear localization of activated MAPKs has been found in all species. Co-localization of these kinases to mitotic microtubules was most evident in L. esculentum, while only about 50% of mitotic cells in the root meristems of P. sativum and V. faba displayed activated MAPKs localized to microtubules during mitosis. Unexpectedly, no evident immunofluorescence signals at spindle microtubules and phragmoplast were noted in L. luteus. Considering immunocytochemical analyses and studies on the impact of FR180204 (an inhibitor of animal ERK1/2) on mitotic cells, we hypothesize that MAPKs may not play prominent role in the regulation of microtubule dynamics in all plant species.

Suppression of the auxin response pathway enhances susceptibility to Phytophthora cinnamomi while phosphite-mediated resistance stimulates the auxin signalling pathway.

BACKGROUND: Phytophthora cinnamomi is a devastating pathogen worldwide and phosphite (Phi), an analogue of phosphate (Pi) is highly effective in the control of this pathogen. Phi also interferes with Pi starvation responses (PSR), of which auxin signalling is an integral component. In the current study, the involvement of Pi and the auxin signalling pathways in host and Phi-mediated resistance to P. cinnamomi was investigated by screening the Arabidopsis thaliana ecotype Col-0 and several mutants defective in PSR and the auxin response pathway for their susceptibility to this pathogen. The response to Phi treatment was also studied by monitoring its effect on Pi- and the auxin response pathways. RESULTS: Here we demonstrate that phr1-1 (phosphate starvation response 1), a mutant defective in response to Pi starvation was highly susceptible to P. cinnamomi compared to the parental background Col-0. Furthermore, the analysis of the Arabidopsis tir1-1 (transport inhibitor response 1) mutant, deficient in the auxin-stimulated SCF (Skp1 - Cullin - F-Box) ubiquitination pathway was also highly susceptible to P. cinnamomi and the susceptibility of the mutants rpn10 and pbe1 further supported a role for the 26S proteasome in resistance to P. cinnamomi. The role of auxin was also supported by a significant (P < 0.001) increase in susceptibility of blue lupin (Lupinus angustifolius) to P. cinnamomi following treatment with the inhibitor of auxin transport, TIBA (2,3,5-triiodobenzoic acid). Given the apparent involvement of auxin and PSR signalling in the resistance to P. cinnamomi, the possible involvement of these pathways in Phi mediated resistance was also investigated. Phi (especially at high concentrations) attenuates the response of some Pi starvation inducible genes such as AT4, AtACP5 and AtPT2 in Pi starved plants. However, Phi enhanced the transcript levels of PHR1 and the auxin responsive genes (AUX1, AXR1and AXR2), suppressed the primary root elongation, and increased root hair formation in plants with sufficient Pi. CONCLUSIONS: The auxin response pathway, particularly auxin sensitivity and transport, plays an important role in resistance to P. cinnamomi in Arabidopsis, and phosphite-mediated resistance may in some part be through its effect on the stimulation of the PSR and auxin response pathways.

Copper microlocalisation and changes in leaf morphology, chloroplast ultrastructure and antioxidative response in white lupin and soybean grown in copper excess.

The microlocalisation of Cu was examined in the leaves of white lupin and soybean grown hydroponically in the presence of 1.6 (control) or 192 µM (excess) Cu, along with its effect on leaf morphology, (ultra)structure and the antioxidative response. The 192 µM dose led to a reduction in the total leaf area and leaf thickness in both species, although more strongly so in white lupin. In the latter species it was also associated with smaller spongy parenchyma cells, and smaller spaces between them, while in the soybean it more strongly reduced the size of the palisade parenchyma and epidermal cells. Energy-dispersive X-ray microanalysis showed that under Cu excess the metal was mainly localised inside the spongy parenchyma cells of the white lupin leaves, and in the lower epidermis cell walls in those of the soybean. Cu excess also promoted ultrastructural chloroplast alterations, reducing the photosynthetic capacity index and the green area of the leaves, especially in the soybean. Despite this, soybean appeared to be more tolerant to Cu excess than white lupin, because soybean displayed (1) lower accumulation of Cu in the leaves, (2) enhanced microlocalisation of Cu in the cell walls and (3) greater levels of induced total -SH content and superoxide dismutase and catalase activities that are expected for better antioxidative responses.

Phenylpropanoid pathway metabolites promote tolerance response of lupine roots to lead stress.

Over the past decade, there has been increasing interest in the role of phenolic compounds, especially flavonoids in plants in response to heavy metal stress. In this study, it was found that treatment of yellow lupine (Lupinus luteus L.) with Pb (150mg/l Pb(NO3)2) increased flavonoid contents in both cotyledons (by ca. 67%) and roots (by ca. 54%). Moreover, seedling roots preincubated with flavonoid extracts, derived from Pb-treated lupine cotyledons, exhibited enhanced tolerance to the heavy metal. Flavonoid preincubated lupine seedlings, growing for 48h in the presence of Pb(NO3)2, showed mitigated symptoms of lead stress, which was manifested by a significant increase in the root length and its biomass. Additionally, in seedlings pretreated with the natural flavonoid preparations an impressive rise of the antioxidant capacity was observed. Simultaneously, root cells exhibited reduced accumulation of both H2O2 and O2(-), which was associated with the decreased TBARS content and the number of dying cells under Pb stress. Taken together, accumulation of flavonoids could be an effective event in the plant׳s spectrum of defense responses to heavy metal stress, and the protective role of flavonoids against heavy metals might be associated with their ability to scavenge reactive oxygen species overproduced under lead stress.

Mitigation of Cu stress by legume-Rhizobium symbiosis in white lupin and soybean plants.

The effect of Bradyrhizobium-legume symbiosis on plant growth, toxicological variables and Cu bioaccumulation was studied in white lupin and soybean plants treated with 1.6, 48, 96 and 192 µM Cu. In both species, those plants grown in the presence of root nodule-forming symbiotic Bradyrhizobium showed less root and shoot growth reduction, plus greater translocation of Cu to the shoot, than those grown without symbiotic Bradyrhizobium. The effective added concentrations of Cu that reduced shoot and root dry weight by 50% (EC50), and the critical toxic concentration that caused a 10% reduction in plant growth (CTC10%), were higher in plants grown with symbiotic Bradyrhizobium, and were in general higher in the roots whether the plants were grown with or without these bacteria. The production of malondialdehyde and total thiols was stimulated by Cu excess in the shoots and roots of white lupin grown with or without symbiotic Bradyrhizobium, but mainly in those without the symbionts. In contrast, in soybean, the increases in malondialdehyde and total thiols associated with rising Cu concentration were a little higher (1.2-5.0 and 1.0-1.6 times respectively) in plants grown with symbiotic Bradyrhizobium than without. Finally, the organ most sensitive to Cu excess was generally the shoot, both in white lupin and soybean grown with or without symbiotic Bradyrhizobium. Further, Bradyrhizobium-legume symbiosis appears to increase the tolerance to Cu excess in both legumes, but mainly in white lupin; plant growth was less reduced and CTC10% and EC50 values increased compared to plants grown without symbiotic Bradyrhizobium. Bradyrhizobium N2 fixation in both legumes would therefore seem to increase the phytoremediation potential of these plants when growing on Cu-contaminated sites.

Phytoremediation of soils co-contaminated by organic compounds and heavy metals: bioassays with Lupinus luteus L. and associated endophytic bacteria.

In the central part of the Iberian Peninsula there are old sealed landfills containing soils co-contaminated by several heavy metals (Cu, Zn, Pb, Cd, Ni, As, Cr, Fe, Al, Mn) and organic pollutants of different families (hydrocarbons, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, pesticides and other organochlorinated compounds, phenols and volatile compounds), which this work will address. We have focused on phytoremedial plants that are able to deal with this type of complex pollution, not only species that tolerate the joint effect of heavy metals in the soil, but also those that can take advantage of associated bacteria to efficiently break down organic compounds. This study was carried out with Lupinus luteus and its endophytes in two greenhouse experiments: A) growing in a substrate artificially contaminated with benzo(a)pyrene (BaP), and B) using real co-contaminated landfill soils. Endophytes of roots and shoots were isolated in both bioassays. Plant growth-promotion tests and organic pollutant tolerance and degradation tests were conducted on all strains isolated in bioassay A), and on those proving to be pure cultures from bioassay B). The selected landfill is described as are isolation and test procedures. Results indicate that plants did not show toxicity symptoms when exposed to BaP but did when grown in landfill soil. Some endophytes demonstrated plant growth-promotion capacity and tolerance to BaP and other organic compounds (diesel and PCB commercial mixtures). A few strains may even have the capacity to metabolize those organic pollutants. The overall decline in plant growth-promotion capacity in those strains isolated from the landfill soil experiment, compared with those from the bioassay with BaP, may indicate that lupin endophytes are not adapted to metal concentration in roots and shoots and fail to grow. As a result, most isolated root endophytes must have colonized root tissues from the soil. While preliminary degradation tests showed promising results (some strains exhibiting the potential to use organic pollutants as their sole source of carbon), these are not conclusive and further in-depth degradation assays need to be performed.

The severity of iron chlorosis in sensitive plants is related to soil phosphorus levels.

BACKGROUND: Iron (Fe) deficiency chlorosis, a major nutritional problem in plants growing on calcareous soils, is related to the content and reactivity of soil iron oxides and carbonates. The effects of other soil components, however, need elucidation. In this paper we tested the hypothesis that application of high doses of phosphorus (P) to the soil can aggravate Fe chlorosis. RESULTS: Lupin and sorghum were grown on 24 calcareous soils. Leaf chlorophyll concentration (LCC) in lupin decreased with increasing available P/available Fe ratio in the native soil but LCC in sorghum was unaffected by that ratio. Application of P to the soil resulted in significant reduction of LCC and dry weight in lupin. In sorghum, LCC and dry weight were positively affected by P fertilisation for soils poor in available P whereas the opposite effect was generally observed for the P-rich soils. In another experiment where olive plants were pot-grown on two soils during the 2009­2011 period, P fertilisation affected LCC negatively only in 2009 and 2011 and in the soil that was poorer in iron oxides. CONCLUSION: Application of fertiliser P to Fe chlorosis-inducing soils is likely to aggravate this deficiency. However, this effect depends on the plant and the Fe and P statuses of the soil.

Root-derived auxin contributes to the phosphorus-deficiency-induced cluster-root formation in white lupin (Lupinus albus).

Formation of cluster roots is a typical morphological response to phosphorus (P) deficiency in white lupin (Lupinus albus), but its physiological and molecular mechanisms are still unclear. We investigated the role of auxin in the initiation of cluster roots by distinguishing the sources of auxin, measuring the longitudinal distribution patterns of free indole-3-acetic acid (IAA) along the root and the related gene expressions responsible for polar auxin transport (PAT) in different developmental stages of cluster roots. We found that removal of shoot apex or primary root apex and application of auxin-influx or -efflux transport inhibitors, 3-chloro-4-hydroxyphenylacetic acid, N-1-naphthylphthalamic acid and 2,3,5-triiodobenzoic acid, to the stem did not affect the number of cluster roots and the free-IAA concentration in the roots of P-deficient plants, but when these inhibitors were applied directly to the growth media, the cluster-root formation was greatly suppressed, suggesting the fundamental role of root-derived IAA in cluster-root formation. The concentration of free IAA in the roots was higher in P-deficient plants than in P-adequate ones, and the highest in the lateral-root apex and the lowest in the mature cluster roots. Meanwhile the expression patterns of LaAUX1, LaPIN1 and LaPIN3 transcripts related to PAT was consistent with concentrations of free IAA along the lateral root, indicating the contribution of IAA redistribution in the cluster-root development. We proposed that root-derived IAA plays a direct and important role in the P-deficiency-induced formation of cluster roots.

White lupin cluster root acclimation to phosphorus deficiency and root hair development involve unique glycerophosphodiester phosphodiesterases.

White lupin (Lupinus albus) is a legume that is very efficient in accessing unavailable phosphorus (Pi). It develops short, densely clustered tertiary lateral roots (cluster/proteoid roots) in response to Pi limitation. In this report, we characterize two glycerophosphodiester phosphodiesterase (GPX-PDE) genes (GPX-PDE1 and GPX-PDE2) from white lupin and propose a role for these two GPX-PDEs in root hair growth and development and in a Pi stress-induced phospholipid degradation pathway in cluster roots. Both GPX-PDE1 and GPX-PDE2 are highly expressed in Pi-deficient cluster roots, particularly in root hairs, epidermal cells, and vascular bundles. Expression of both genes is a function of both Pi availability and photosynthate. GPX-PDE1 Pi deficiency-induced expression is attenuated as photosynthate is deprived, while that of GPX-PDE2 is strikingly enhanced. Yeast complementation assays and in vitro enzyme assays revealed that GPX-PDE1 shows catalytic activity with glycerophosphocholine while GPX-PDE2 shows highest activity with glycerophosphoinositol. Cell-free protein extracts from Pi-deficient cluster roots display GPX-PDE enzyme activity for both glycerophosphocholine and glycerophosphoinositol. Knockdown of expression of GPX-PDE through RNA interference resulted in impaired root hair development and density. We propose that white lupin GPX-PDE1 and GPX-PDE2 are involved in the acclimation to Pi limitation by enhancing glycerophosphodiester degradation and mediating root hair development.

Metabolic analysis revealed altered amino acid profiles in Lupinus albus organs as a result of boron deficiency.

We analysed the changes in the metabolites of Lupinus albus organs (leaf-blades, petioles, apexes, hypocotyls and roots) as a consequence of B deficiency. The deficiency did not affect malate concentration and induced only minor changes in the sugar content, suggesting that the carbohydrate metabolism is little affected by the deficiency. Contrarily, marked changes in the content of free amino acids were observed, with some specific variations associated with the different organs. These changes indicate that various aspects of metabolism implicated in the amino acid accumulation were affected by B deficiency. Most of the detected changes appear to have implications with some stress responses or signalling processes. Asparagine and proline that increase in many stresses also accumulated in petioles, apexes and hypocotyls. Accumulation of γ-aminobutyric acid shunt amino acids, indicative of production of reactive oxygen species, occurs in the same three organs and also the roots. The increase in the branched-chain amino acids, observed in all organs, suggests the involvement of B with the cytoskeleton, whereas glycine decrease in leaf-blades and active growing organs (apexes and roots) could be associated with the proposed role of this amino acids in plant signalling in processes that might be associated with the decreased growth rates observed in B deficiency. Despite the admitted importance of free amino acids in plant metabolism, the available information on this matter is scarce. So our results bring new information concerning the effects of B deficiency in the metabolism of the several L. albus organs.

Localized application of soil organic matter shifts distribution of cluster roots of white lupin in the soil profile due to localized release of phosphorus.

BACKGROUND AND AIMS: Phosphorus (P) is a major factor controlling cluster-root formation. Cluster-root proliferation tends to concentrate in organic matter (OM)-rich surface-soil layers, but the nature of this response of cluster-root formation to OM is not clear. Cluster-root proliferation in response to localized application of OM was characterized in Lupinus albus (white lupin) grown in stratified soil columns to test if the stimulating effect of OM on cluster-root formation was due to (a) P release from breakdown of OM; (b) a decrease in soil density; or (c) effects of micro-organisms other than releasing P from OM. METHODS: Lupin plants were grown in three-layer stratified soil columns where P was applied at 0 or 330 mg P kg(-1) to create a P-deficient or P-sufficient background, and OM, phytate mixed with OM, or perlite was applied to the top or middle layers with or without sterilization. KEY RESULTS: Non-sterile OM stimulated cluster-root proliferation and root length, and this effect became greater when phytate was supplied in the presence of OM. Both sterile OM and perlite significantly decreased cluster-root formation in the localized layers. The OM position did not change the proportion of total cluster roots to total roots in dry biomass among no-P treatments, but more cluster roots were concentrated in the OM layers with a decreased proportion in other places. CONCLUSIONS: Localized application of non-sterile OM or phytate plus OM stimulated cluster-root proliferation of L. albus in the localized layers. This effect is predominantly accounted for by P release from breakdown of OM or phytate, but not due to a change in soil density associated with OM. No evidence was found for effects of micro-organisms in OM other than those responsible for P release.

Efficiency of white lupin in the removal of mercury from contaminated soils: soil and hydroponic experiments.

This study examined the ability of the white lupin to remove mercury (Hg) from a hydroponic system (Hg concentrations 0, 1.25, 2.5, 5 and 10 micromol/L) and from soil in pots and lysimeters (total Hg concentration (19.2 +/- 1.9) mg/kg availability 0.07%, and (28.9 +/- 0.4) mg/kg availability 0.09%, respectively), and investigated the accumulation and distribution of Hg in different parts of the plant. White lupin roots efficiently took up Hg, but its translocation to the harvestable parts of the plant was low. The Hg concentration in the seeds posed no risk to human health according to the recommendations of the World Health Organization, but the shoots should not be used as fodder for livestock, at least when unmixed with other fodder crops. The accumulation of Hg in the hydroponically-grown plants was linear over the concentration range tested. The amount of Hg retained in the roots, relative to the shoots, was almost constant irrespective of Hg dose (90%). In the soil experiments, Hg accumulation increased with exposure time and was the greater in the lysimeter than in the pot experiments. Although Hg removal was the greater in the hydroponic system, revealing the potential of the white lupin to extract Hg, bioaccumulation was the greatest in the lysimeter-grown plants; the latter system more likely reflects the true behaviour of white lupin in the field when Hg availability is a factor that limits Hg removal. The present results suggest that the white lupin could be used in long-term soil reclamation strategies that include the goal of profitable land use in Hg-polluted areas.