Aerobic and anaerobic nitrate and nitrite reduction in free-living cells of Bradyrhizobium sp. (Lupinus)

Induction, energy gain, effect on growth, and interaction of nitrate and nitrite reduction of Bradyrhizobium sp. (Lupinus) USDA 3045 were characterized. Both nitrate and nitrite were reduced in air, although nitrite reduction was insensitive to ammonium inhibition. Anaerobic reduction of both ions was shown to be linked with energy conservation. A dissimilatory ammonification process was detected, which has not been reported in rhizobia so far. Nevertheless, anaerobic conversion of nitrate to ammonium was lower than 40%, which suggests the presence of an additional, nitrite reductase of denitrifying type. Nitrite toxicity caused a non-linear relationship between biomass produced and >2 mM concentrations of each N oxyanion consumed. At > or =5 mM initial concentrations of nitrate, a stoichiometric nitrite accumulation occurred and nitrite remained in the medium. This suggests an inhibition of nitrite reductase activity by nitrate, presumably due to competition with nitrate reductase for electron donors. Lowering of growth temperature almost completely diminished nitrite accumulation and enabled consumption as high as 10 mM nitrate, which confirms such a conclusion.     

Potassium nutrition effects on seed alkaloid concentrations,yield and mineral content of lupins (Lupinus angustifolius)

To ensure that narrow-leafed lupin (Lupinus angustifolius L.) meets feed quality standards, the concentration of alkaloids must be kept under the maximum acceptable limit of 200 mg kg^–1 DM. One of the factors that may affect seed alkaloid concentration is soil nutrient deficiency. In this paper, we report the results of glasshouse and field experiments that tested the effect of potassium (K) deficiency on seed alkaloid concentrations. In the glasshouse, seed alkaloid concentrations increased by 385, 400 and 205% under severe K deficiency in sweet varieties (Danja, Gungurru and Yorrel, respectively) of L. angustifolius. The concentration of alkaloids in Fest, the bitter variety, was always high regardless of soil K status. At all levels of applied K (0–240 mg kg^–1 soil), lupanine was the predominant alkaloid in sweet varieties, whereas 13-hydroxylupanine prevailed in the bitter variety. Seed yield of all varieties increased exponentially with increasing amounts of applied K, reaching a maximum at 60 mg K kg^–1 soil. In the field, application of K to deficient soils decreased seed alkaloid concentration at Badgingarra, Western Australia (WA) but not at Nyabing, WA, in 1996. In both field trials, seed yield and mineral content were not affected by the amounts of K fertiliser applied. These findings highlighted the need for adequate K fertilisation of deficient soils in WA to avoid the risk of producing low quality lupin seed with high alkaloid concentrations. K deficiency is involved in stimulating alkaloid production in sweet varieties of L. angustifolius.     

Purification of a cytochrome aa3 terminal oxidase from protoplast membrane vesicles of Micrococcus luteus

Abstract A cytochrome aa₃ terminal oxidase was isolated from protoplast membrane vesicles of Micrococcus luteus grown under aerobic conditions. The purified complex showed similarities to cytochrome c oxidase (EC 1.9.3.1) of the electron transport chain of mitochondria and many prokaryotes. The enzyme was solubilized by subsequent treatment with the detergents CHAPS and n-dodecyl-β-d-maltoside and purified by ion-exchange chromatography using poly-L-lysine agarose and TMAE-fractogel-650 (S) columns, followed by hydroxyapatite chromatography. The purified complex is composed of two major subunits with apparent molecular masses of 54 and 32 kDa. After purification the isolated enzyme contains 12.1 nmol of heme A (mg protein)⁻¹ and exhibits absorption maxima at 424 nm and 598 nm in the oxidized state and at 442 nm and 599 nm in the reduced state. The CO-difference spectrum shows peaks at 428 and 590 nm which is indicative of heme a ₃, furthermore oxygen consumption was found to be sensitive to cyanide.     

The acquisition of phosphorus byLupinus albus L.

Summary It has been demonstrated by an agar film technique thatL. albus can cause the breakdown of colloids of iron/silicate, iron/phosphate, aluminium/silicate and aluminium phosphate and destabilise suspensions of manganese dioxide, calcium mono-hydrogen phosphate and ferric hydroxide. Dissolution of these compounds was most marked in areas adjacent to proteoid roots (dense clusters of secondary laterals of limited growth which develop on lateral roots) and parts of the tap root. Soil associated with these regions of the root system contained more reductants and chelating agents than the bulk soil. Soil from around the roots ofL. albus exhibited much greater reducing and chelating activity than that associated with the roots of rape and buckwheat.     

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.     

The maximum axial growth pressure of roots of spring and autumn cultivars of lupin

The maximum axial growth pressure of roots of Lupinus albus cv Lublanc and Lupinus albus cv Lunoble (spring and autumn-sown cultivars respectively) were measured. The mean values were not significantly different with an overall mean value of 645 kPa. This value is not unusually large for plant roots and is surprising because lupins are known for their ability to penetrate strong soils. The autumn cultivar had a significantly greater maximum elongation rate under zero mechanical impedance than the spring cultivar. The impeded diameters were also larger in the autumn cultivars.     

How does insect visitation trigger floral colour change?

Abstract.  1. Visitation by the key pollinator, Bombus terrestris , was implicated in inducible flower colour change in Lupinus pilosus . Behaviour at the flower and rate of visitation by these bumble bees had specific effects; exclusion of this flower visitor led to retarded onset, and reduced rate, of colour change.
2. The foraging behaviour of B. terrestris was influenced by floral colour change in L. pilosus . Choice of pre-change flowers was greater than random in relation to the proportion of colour phases available within the plant population.
3. Levels of floral manipulation that mimicked the flower handling characteristics of visiting bumble bees confirmed that triggering of the pollen release mechanism is necessary for the instigation of colour change.
4. Moreover, this study suggests that, in L. pilosus , an aspect of pollination (pollen deposition by bees and/or subsequent pollen tube growth within the style) is linked with colour change and may act as the trigger for such change.     

Development and implementation of a sequence-specific PCR marker linked to a gene conferring resistance to anthracnose disease in narrow-leafed lupin (Lupinus angustifolius L.)

Anthracnose caused by Colletotrichum gloeosporioides is the most serious disease of lupins (Lupinus spp). A cross was made between cultivars Tanjil (resistant) and Unicrop (susceptible) in narrow-leafed lupin (L. angustifolius). Analysis of disease reaction data on the F₂ population and on the resultant F₇ recombinant inbred lines suggested that Tanjil contained a single dominant gene (Lanr1) conferring resistance to anthracnose. The parents and the representative F₂ plants were used to generate molecular markers liked to the Lanr1 gene using the MFLP technique. A co-dominant MFLP polymorphism linked to the Lanr1 gene was identified as a candidate marker. The bands were isolated, re-amplified by PCR, cloned and sequenced. The MFLP polymorphism was converted into a co-dominant, sequence-specific, simple PCR-based marker. Linkage analysis by the computer program MAPMAKER indicated that the marker was 3.5 centiMorgans (cM) from the gene Lanr1. This marker is currently being implemented for marker assisted selection in the Australian National Lupin Breeding Program.     

Über polykomplexartige Strukturen in den Kernen vonLupinus polyphyllus

In electron micrographs ofLupinus polyphyllus nuclei some of the chromocentres exhibit two distinct regions, a network-like (NR) and a banded one (GR). This heterogeneous type of chromocentres was observed in nearly all cells of vegetative and reproductive tissues (root, leaf primordium of the seedling, young and older foliage leaves, carpel, young and older ovules and pericarp), with the exception of pollen mother cells during the first meiotic division. Results of enzymatic digestion show that GR and NR mainly consist of DNA. These observations suggest that GR and the polycomplexes represent different nuclear phenomena.

Herrn Univ.-Prof. Dr.Walter Leinfellner zum 70. Geburtstag gewidmet.     

Cluster-root production and organic anion exudation in a group of old-world lupins and a new-world lupin

We examined the capacity of several Old-World lupin species (Lupinus luteus L., L. hispanicus Boiss. et Reuter and L. angustifolius L.) and one species of a New-World lupin (L. mutabilis Sweet) to form cluster roots under a range of conditions in solution culture. The effect of the synthetic auxin, IBA (indole-3-butyric acid), on cluster-root development in L. luteus and L. albus L. provided with an adequate phosphorus (P) supply was also investigated. In addition, the effect of a high nitrate-N (NO₃-N) supply on the efflux of citrate and malate from roots of L. angustifolius was examined to determine if specific regions of the root system exuded these organic anions. When P-deficient, L. hispanicus, L. luteus and L. mutabilis formed cluster roots that secreted organic anions. Citrate was generally the dominant organic anion exuded, although succinate was also exuded in large quantities from L. luteus. Citrate efflux by L. hispanicus and L. luteus was at least comparable to that reported for P-deficient L. albus[up to 1.092 nmol g^–1 fresh weight (FW) s^–1], but was over an order of magnitude lower in L. mutabilis (0.036 nmol g^–1 FW s^–1). Citrate and malate were not detected in significant amounts from either the lateral roots or the root tips of any species grown under P-sufficient or -deficient conditions. Citrate efflux from cluster roots of L. luteus showed a diurnal pattern, similar to that reported for L. albus, with maximum efflux during the day, and declining to a minimum before dawn. IBA added to the nutrient solution induced cluster-root formation on both L. albus and L. luteus at concentrations of P that would normally suppress the production of these roots. However, the IBA-induced cluster roots did not exude significant amounts of citrate. Although L. angustifolius did not produce cluster roots when P-deficient, it produced cluster-like root structures that exuded citrate (0.053 nmol g^–1 FW s^–1) when grown at a high nitrate-N (NO₃-N) supply. L. angustifolius did not exude significant citrate or malate from lateral roots or root tips when grown at either high or low NO₃-N supply. Our findings for L. hispanicus and L. luteus are the first reports of cluster-root formation in response to P deficiency for these Old-World species, and for L. mutabilis, it is the first report of cluster roots for a New-World lupin species. These reports indicate that evolutionary and biogeographical aspects of cluster-root formation in the genus Lupinus need to be revised. Furthermore, investigation is warranted to determine the capacity of species of the large group of New-World lupins to form cluster roots in soils of their native habitats.