Yellow Lupin Roots Research Articles

Carbon and nitrogen mineralisation from green manures as alternative nitrogen sources in Mediterranean farming

ABSTRACTThe soil incorporation of green manures is a practice that can be used in sustainable agriculture and in organic farming, where nitrogen (N) sources are limited. The aim of this study was to evaluate balansa clover (Trifolium michelianum Savi), yellow lupine (Lupinus luteus L.) and ryegrass (Lolium multiflorum Lam.) as potential alternative N sources. A total of nine treatments were considered in this study: control, aerial of balansa clover, roots of balansa clover, aerial of yellow lupine, roots of yellow lupine, aerial of ryegrass, roots of ryegrass, mixture aerial + roots of yellow lupine and mixture aerial + roots of ryegrass. A laboratory incubation experiment was conducted under controlled conditions during 196 days and carbon and N mineralisation were followed. Results showed that green manures are appropriate N sources for Mediterranean farming. No significant differences in terms of N mineralisation were observed between aerial or roots biomass of the green manures. Besides, 37–55% of total N applied was mineralised in treatments amended with balansa clover or yellow lupine, whereas 13–21% of total N applied was mineralised in ryegrass. It can be concluded that the most efficient green manure for supplying mineral N to the succeeding crop was yellow lupine.

Effect of neutralizing substances on zinc-contaminated soil on the yield and macroelement content in yellow lupine (Lupinus luteus L.)

Some trace elements, for example zinc, play both a positive and a negative role in plant life, which requires their content in soil. If soil is excessively contaminated with zinc, an attempt should be made to reduce the negative effect of this element on plants and other living organisms. For this reason, a study was undertaken to determine whether it was possible to alleviate the effect of soil zinc contamination (0, 150, 300 and 600 mg Zn kg-1 of soil) on the yield and macronutrient content of yellow lupine (Lupinus luteus L.). Compost (3%), bentonite (2%) and zeolite (2% relative to soil mass) were used to reduce the effect of soil zinc contamination. Macroand micronutrients were applied to the soil in the same amounts in all pots: 30 mg N, 30 mg P, 100 mg K, 50 mg Mg, 0.33 mg B, 5 mg Mn and 5 mg Mo per kg soil. Yellow lupine was harvested in the flowering phase and plant material samples were collected for laboratory tests. The induced soil zinc contamination reduced yellow lupine growth and development because a dose of 300 mg Zn kg-1 soil caused plant seedlings to wither. Compost and bentonite reduced the negative influence of soil zinc contamination on yellow lupine yield, especially on aerial parts. The most demonstrable effect of zinc on the macronutrient content of lupine plants was recorded for magnesium and calcium, whose content increased compared to the control in both the aerial parts and roots of yellow lupine. Among the neutralizing substances, the effect of zeolite on the phosphorus, magnesium and calcium content and bentonite on the sodium content in the plants was the most beneficial.

Water status and water diffusion transport in lupine roots exposed to lead

Water status and diffusion transport were studied in the roots of yellow lupine (Lupinus luteus L., cv. Juno) treated for 48h with two selected concentrations of Pb(NO3)2: 150mgl−1, which inhibited root growth by about 50% (medium stress intensity), as well as 350mgl−1, which almost entirely suppressed root elongation (severe stress intensity). Relative water content (RWC), which characterizes the degree of root water saturation, slightly increased at the lower lead concentration and remained unchanged at the higher lead dose. Ultrastructure analyses under a transmission electron microscope revealed that plasmolysis was not evoked by lead in the apical part of the meristem. Moreover, direct observation of meristem cells using Nomarsky optics indicated enhanced vacuolization in the presence of both lead concentrations. These data suggest that the water status of the roots was not affected by the metal. Due to the fact that proline is involved in the maintenance of turgor in the cells, the metabolism of this amino acid was investigated. In the roots, the activity of enzymes involved in proline synthesis, such as pyrroline-5-carboxylate synthetase (P5CS) and ornithine aminotransferase (OAT), increased at 150mgl−1 Pb2+; nevertheless, proline content was diminished at the lower lead concentration. This effect is likely the result of proline degradation by proline dehydrogenase (PDH), since the activity of this enzyme increased at the lower lead dose. On the other hand, in the presence of 350mgl−1 Pb2+, a low level of proline was correlated with a decrease in the activity of P5CS and OAT, as well as unchanged PDH activity in lupine roots. These data may imply that enzymatic synthesis of proline was strongly damaged by the metal ions. The low level of proline in both experimental variants suggests that proline accumulation is inessential to maintaining the osmotic uptake of water into root cells. NMR spectroscopy showed that exposition of lupine seedlings to lead caused a deceleration in water transport in the roots due to a reduction in the water transfer rate across the membranes (transmembrane transfer) and vacuoles continuum, as well as water diffusion along the root apoplast. Fluorescence staining and immunogold labeling showed the presence of callose strands in cell walls and/or in the vicinity of them. In lead-treated lupine roots, callose was mainly localized in the parenchyma cortex placed lengthwise to the vascular cylinder. Callose deposits in the cell walls may reduce vacuolar transport, as well as increase cell wall resistance to water flow. Deceleration of diffusional water movement to the vascular system, may in turn, influence the rate of long-distance water transport to aerial parts of the plant.

Susceptibility of common herbaceous crops to Phytophthora cinnamomi and its influence on Quercus root rot in rangelands

The susceptibility to Phytophthora cinnamomi of four crops (wheat, oat, vetch, and yellow lupin) commonly planted in rangeland ecosystems in southern Spain was evaluated. By means of in vitro infection experiments, the presence of the pathogen into the roots of yellow lupin (symptomatic) and vetch (asymptomatic) was observed, but never into wheat and oat roots (asymptomatic). It was also demonstrated that yellow lupin stimulated the production of zoospores of P. cinnamomi. Vetch, wheat and oat did not stimulate zoospore production. Under controlled conditions, only yellow lupin induced an increase in the number of viable chlamydospores in the soil. We concluded that the culture of wheat, oat, and vetch in rangelands did not influence the epidemiology of the Quercus root disease, even when asymptomatically-infected vetch is grown, and these crops can constitute an alternative to the culture of yellow lupin in rangeland ecosystems affected by Quercus root rot.

Effect of soil contamination with arsenic and application of different substances on the manganese content in plants

The aim of the study has been to determine the effect of some substances such as dolomite, loam, compost, pinewood bark, peat, lime, charcoal, natural and synthetic zeolite on reducing the impact of soil contamination with arsenic on the content of manganese in some plant species. The content of manganese in the test plants depended on the degree of soil contamination with arsenic, application of different substances as well as on the plant species and organ. Soil contamination with arsenic caused either an increase or a decrease in the content of manganese in plants depending on a plant species and organ. In the series without soil amending substances, in the arsenic contaminated objects the manganese content decreased in above-ground parts of cocksfoot and swede but increased in above-ground parts and roots of maize and yellow lupine, in roots of cocksfoot and swede and in straw and roots of spring barley. On the other hand, the highest rates of arsenic depressed the content of manganese in roots of cocksfoot, swede and spring barley. Addition of any of the aforementioned substances to contaminated soil changed the content of manganese in the plants. The most unambiguous effect of the different substances was determined in the case of above-ground parts of maize as well as above-ground parts and roots of cocksfoot, in which the manganese content fell down, and in roots of yellow lupine, grain and straw of spring barley, in which the content of manganese rose. Charcoal and loam caused the largest and synthetic zeolite led to the smallest changes in the content of manganese in plants.

Organ-specific expression of glutamate dehydrogenase (GDH) subunits in yellow lupine

Glutamate dehydrogenase (GDH, EC 1.4.2-4) is present in yellow lupine ( Lupinus luteus cv. Juno) in many isoforms. The number and banding pattern of isoenzymes varies with respect to plant organ and developmental stage. To better understand the complex nature of GDH regulation in plants, the levels of GDH transcripts, enzyme activity and isoenzyme patterns in germinating seeds and roots of yellow lupine were examined. The analysis of GDH cDNA sequences in lupine revealed three mRNA types, of which two encoded the β-GDH subunit and one encoded the α-GDH subunit (corresponding to the GDH1( GDH3) and GDH2 genes, respectively). The relative expression of GDH1 and GDH2 genes was analyzed in various lupine organs by using quantitative real-time PCR. Our results indicate that different mRNA types were differently regulated depending on organ type. Although both genes appeared to be ubiquitously expressed in all lupine tissues, the GDH1 transcripts evidently predominated over those of GDH2. Immunochemical analyses confirmed that, during embryo development, varied expression of two GDH subunits takes place. The α-GDH subunit (43 kDa) predominated in the early stages of germinating seeds, while the β-GDH subunit (44 kDa) was the only GDH polypeptide present in lupine roots. These results firmly support the hypothesis that isoenzyme variability of GDH in yellow lupine is associated with the varied expression of α and β subunits into the complexes of hexameric GDH forms. The presence of several isogenes of GDH in yellow lupine may explain the high number (over 20) of its molecular forms in germinating lupine.

Effect of soil contamination with diesel oil on yellow lupine yield and macroelements content

The study has been undertaken to assay the effect of soil contamination with diesel oil on yellow lupine yield and macroelement contents as well as to examine the relationships between the accumulation of macroelements versus the yellow lupine yielding and some yield quality characteristics. The influence of soil pollution with refinery products depended on the type of soil, diesel oil concentration in soil and mineral fertilisation. Increasing contents of soil pollution with diesel oil promoted higher concentrations of phosphorus, sodium, magnesium and calcium in aboveground parts and roots of yellow lupine. More severe contamination was also responsible for depressed yield of yellow lupine aboveground parts and roots as well as accumulation of nitrogen in aboveground parts and potassium in roots of the crop. The addition of nitrogen to soil diversified the yielding and content of macroelements in yellow lupine. The direction these changes took depended on the plant organ, type of soil and degree of diesel oil contamination.

Calcium Status of Yellow Lupin Symbiosomes as a Potential Regulator of Their Nitrogenase Activity: The Role of the Peribacteroid Membrane

The capacity of symbiosomes from yellow lupin root nodules for active Ca2+uptake and the sensitivity of their nitrogenase activity to a disturbance of the symbiotic Ca partition were investigated. The experiments carried out on the isolated symbiosomes and the peribacteroid membrane (PBM) vesicles, using Ca2+indicators arsenazo III and chlorotetracycline, and the cytochemical Ca visualization with potassium pyroantimonate (PA) provided evidence that an Mg-ATP-energized pump, most likely Mg2+-dependent Ca2+-ATPase catalyzing the active transport of Ca2+from the cytosol of the plant cell into the symbiosomes across the PBM, functions on this membrane. Depleting the symbiosomes of Ca both in vivoandin vitroby treating the intact nodules of yellow lupin root or the purified symbiosomes isolated from the latter with EGTA and Ca2+-ionophore A23187 substantially decreased their nitrogenase activity. The inhibitory effect of calcium deficit in the symbiosomes was not reversed by the addition of calcium to the incubation medium containing the plant tissues under study and was even enhanced under these conditions. The nitrogenase activity of the isolated symbiosomes not experiencing calcium deficit was also inhibited by the addition of relatively high concentrations of exogenous calcium to the incubation medium. These results seem to give evidence that the calcium status of nodule symbiosomes from yellow lupin roots controls their nitrogenase activity. The data obtained suggest that both Ca2+transport on PBM and the low passive permeability of this membrane for the given cation play the key role in such a control.

Characterization and expression analysis of the yellow lupin (Lupinus luteus L.) gene coding for nodule specific proline-rich protein.

The LlPRP2 gene coding for a proline-rich protein shows a high level of similarity to, as well as significant differences from the family of ENOD2 nodule-specific genes. Several sequence motifs with putative regulatory function were identified in the 5' and 3' noncoding regions of the LlPRP2 gene. Northern blot analysis revealed that the expression of the LlPRP2 gene begins 9 days after inoculation of yellow lupin roots with Bradyrhizobium sp. (Lupinus); the expression is restricted to symbiotic nodules and is not detected in other tissues or organs. Detailed hybridization analysis showed that, when expression is activated, the LlPRP2 transcript is modified so as to produce at least three bands and a continuous distribution of decay intermediates. The modification of the LlPRP2 transcript probably involves degradation from the 5'- and/or 3'-ends of the RNA molecules. Southern blot analysis indicates that only one gene is present in the yellow lupin genome. The presence of genes homologous to the LlPRP2 gene was confirmed for three cultivars of yellow lupin and for Lupinus angustifolius. However, LlPRP2 homologues were not detected in Lupinus albus cv. Bac, indicating that this plant may lack the ENOD2 sequence.

Yellow lupin (Lupinus luteus) tolerates waterlogging better than narrow-leafed lupin (L. angustifolius) III. Comparison under field conditions

This study determined whether the tolerance of yellow lupin to waterlogging, observed in experiments in controlled environments, occurs under field conditions. Of particular interest is the impact of waterlogging on the distribution of roots because lupin is exposed to terminal drought in the south of Western Australia, which in itself can have a profound effect on yield. A field experiment was undertaken in the central grain-growing region of Western Australia near Beverley using hydraulically isolated plots to impose and remove waterlogging in a duplex soil. The responses of root and shoot growth of narrow-leafed and yellow lupin to waterlogging in the field were similar to those observed in the controlled environment experiments. In the field experiment, waterlogging had no effect on seed yield of yellow lupin but reduced it by 61% in narrow-leafed lupin. Waterlogging more than halved the dry weight of narrow-leafed lupin but reduced it by only 19% in yellow lupin. In yellow lupin, yield was 3.4 t/ha with waterlogging and 3.8 t/ha without waterlogging, compared with 1.4 t/ha with waterlogging and 3.5 t/ha without waterlogging in narrow-leafed lupin. Waterlogging had no effect on the harvest index of yellow lupin (0.26) but reduced it from 0.36 to 0.31 in narrow-leafed lupin. The larger effect of waterlogging on the yield of narrow-leafed lupin was mainly attributable to fewer pods. Net root growth ceased during waterlogging in both species. After waterlogging, roots of yellow lupin grew at a similar rate to the controls, whereas roots of narrow-leafed lupin grew at a much slower rate than the controls. Waterlogging halved the root density of yellow lupin at 25 cm depth and almost eliminated the roots of narrow-leafed lupin at this depth. After waterlogging, root production in the surface 10 cm increased to about 0.5 cm/cm 3 in yellow lupin but to 0.2 cm/cm 3 in narrow-leafed lupin. At depth (>20 cm), roots of waterlogged yellow lupin continued to grow while those of waterlogged narrow-leafed lupin grew little, if at all. Yellow lupin tolerated waterlogging in the field better than narrow-leafed lupin because it re-established its root system at depth after waterlogging was removed and it produced more fertile pods.

Heavy Metal-induced Polypeptides in Lupin Roots are Similar to Pathogenesis-related Proteins

Heavy metals, such as lead, copper and cadmium, induced in yellow lupin roots the appearance of polypeptides of MW about 16kDa (Przymusiński and Gwózdó, 1994). It was shown that this fraction, migrating on 12.5% (w/v) denaturing polyacrylamide gels as a single broad band, is in fact composed of six components, differing only slightly in their electrophoretic mobility. The sequence analysis ofN-terminal amino acids indicated that all polypeptides composing the 16kDa fraction, showed high degree of homology to the PR-10 family of pathogenesis-related proteins, and among them the polypeptide designated PR-L1 is most related to the PR proteins. Some of those lupin proteins display also a sequence homology (50-55%) to tree pollen allergens and RNase from ginseng callus. In addition to PR-like proteins, the cytosolic Zn, Cu-superoxide dismutase was also detected within 16 kDa protein fraction. The location of the enzyme partially corresponds to the fastest migrating PR-Ls polypeptides (PR-L5 and PR-L6), however the majority of SOD protein was localized below the PR-Ls polypeptide bands.

Prenylated flavonoids in the roots of yellow lupin

A further investigation of the methanol-soluble compounds in yellow lupin roots has revealed a new diprenylchromone, a new coumaronochromone (lupinalbin H), a new isoflavone 5,7,4′-trihydroxy-8,3′-di-(3,3-dimethylallyl)isoflavone(isolupalbigenin), and some complex flavanones. The latter compounds have been identified as two known diprenylated flavanones (lonchocarpol A and euchrestaflavanone A), two diasteroisomeric pairs of dihydrofuranoflavanones (lonchocarpols C 1 and C 2, and lonchocarpols D 1 and D 2; the structures formerly proposed for lonchocarpols C and D were also reinvestigated), a new furanoflavanone (lupinenol), and three 8-prenylflavanones with an additional (2 RS)-hydroxy-3-methyl-3-butenyl side chain. The structures of the latter flavanones were unambiguously identified by spectroscopic ( 1H NMR) comparison with 6-, 8- and 3′-prenylnaringenins chemically prepared from (2 S)-naringenin. The antifungal activity of the prenylated naringenins, and of the various yellow lupin flavanones, was determined by TLC place bioassays using Cladosporium herbarum as the test fungus.

Partial purification, characterization, and kinetic analysis of isoflavone 5- O-methyltransferase from yellow lupin roots

An isoflavone 5- O-methyltransferase was partially purified from the roots of yellow lupin ( Lupinus luteus) by fractional precipitation with ammonium sulfate, followed by gel filtration and ion-exchange chromatography using a fast-protein liquid chromatography system. This enzyme, which was purified 810-fold, catalyzed position-specific methylation of the 5-hydroxyl group of a number of substituted isoflavones. The methyltransferase had a pH optimum of 7 in phosphate buffer, an apparent pI of 5.2, a molecular weight of 55,000, no requirement for Mg 2+, and was inhibited by various SH-group reagents. Substrate interaction kinetics of the isoflavonoid substrate and S-adenosyl- l-methionine gave converging lines which were consistent with a sequential bireactant binding mechanism. Furthermore, product inhibition studies showed competitive inhibition between S-adenosyl- l-methionine and S-adenosyl- l-homocysteine and noncompetitive inhibition between the isoflavone and either S-adenosyl- l-homocysteine or the 5- O-methylisoflavone. The kinetic patterns obtained were consistent with an ordered bi bi mechanism, where S-adenosyl- l-methionine is the first substrate to bind to the enzyme and S-adenosyl- l-homocysteine is the final product released. The physiological role of this enzyme is discussed in relation to the biosynthesis of 5- O-methylisoflavones of this tissue.

New 3-methoxyflavones in the roots of yellow lupin (Lupinus luteus L. cv. Topaz).

An isopropenyl ( = 3,3-dimethylallyl) 3-methoxyflavone (1) and its hydrate (5) were isolated from the roots of yellow lupin, Lupinus luteus L. cv. Topaz. Their structures were unambiguously determined to be 5,7,4′-trihydroxy-3-methoxy-6-(3,3-dimethylallyl)flavone (1) and 5,7,4′-trihydroxy-6-(3-hydroxy-3-methylbutyl)-3-methoxyflavone (5) by a combination of chemical and spectroscopic methods, and the new flavones were named topazolin and topazolin hydrate, respectively.Antifungal tests against the growth of Cladosporium herbarum indicated that, in spite of its phenolic nature and the possession of an isopentenyl sidechain, topazolin (1) had only weak fungitoxic activity.

Isoflavonoids of yellow lupin. Part I. New complex isoflavones in the roots of yellow lupin (Lupinus luteus L., cv. Barpine).

Six new isoflavones each having a prenyl (3, 3-dimethylallyl) or a modified prenyl group at C-6 or C-8 of the isoflavone skeleton, lupiwighteone [5, 7, 4'-trihydroxy-8-(3, 3-dimethylallyl)isoflavone, 1], luteone hydrate [5, 7, 2', 4'-tetrahydroxy-6-(3-hydroxy-3-methylbutyl)isoflavone, 3 (already known as a fungal metabolite of luteone)], lupiwighteone hydrate [5, 7, 4'-trihydroxy-8-(3-hydroxy3-methylbutyl)isoflavone, 4], 2, 3-dehydrokievitone hydrate [5, 7, 2', 4'-tetrahydroxy-8-(3-hydroxy3-methylbutyl)isoflavone, 5], hydroxywighteone [5, 7', 4'-trihydroxy-6-(3-hydroxymethyl-3-methylallyl)isoflavone, 6], hydroxyparvisoflavone A [a 2-hydroxymethyl-2-methylpyrano-isoflavone, 7], and a new coumaronochromone, lupilutin [5, 7, 4'-trihydroxy-8-(3-hydroxy-3-methylbutyl)-coumaronochromone, 8], were isolated from the roots of yellow lupin (Lupinus luteus L., cv. Barpine). Together with these new isoflavonoids, the presence of simple isoflavones (genistein and 2'-hydroxygenistein) and complex isoflavones (wighteone, luteone, 2, 3-dehydrokievitone (2), parvisoflavones A and B, and lupinisoflavones A and B) in the yellow lupin roots has also been confirmed.

Desaturation of oleoyl and linoleoyl residues linked to phospholipids in growing roots of yellow lupin

When growing lupin roots were fed with [ 14C]oleate and [ 14C]linoleate most of the absorbed molecules remained in the free fatty acid pool. Part of the 14C-labelled fatty acid was integrated first into phosphatidylcholine and subsequently into phosphatidylethanolamine. Then, progressive desaturation took place. The kinetics of these processes suggests that desaturation in phosphatidylcholine was followed by transacylation of polyunsaturated fatty acids into phosphatidylethanolamine. To produce good yields of desaturation (about 48%), seeds had to be less than 1-year old. With respect to linoleate desaturation, seedlings had to be 2–7-days old. Increased percentages of oleate desaturation were obtained when the roots were grown on a medium enriched with calcium.

Localization of Glutamate Dehydrogenase and Glutamate Synthase in Roots and Nodules of Lupinus Seedlings

Activities of glutamate dehydrogenase (EC 1.4.1.2. GDH) and glutamate synthase (EC 1.6.1.53. GOGAT) in bacteroids, mitochondria, and cytosol of root nodules, as well as in mitochondria and cytosol of roots of yellow lupine were studied. A high NADH-dependent GDH activity was proved in the nodules, localized mainly in the mitochondrial fraction. In bacteroids and cytosol of the nodules, the NADH-dependent GOGAT activity exceeded the GDH activity. Both GDH and GOGAT revealed a significantly higher activity with NADH than in the presence of NADPH. Histochemical studies demonstrated the highest activity of glutamate dehydrogenase in the roots of lupine in the cells surrounding the nodule. Preliminary electron-microscopic investigations showed that in these cells two morphological types, at least, of mitochondria are apparent.