Endosymbiotic bacteria nodulating a new endemic lupine Lupinus mariae-josephi from alkaline soils in Eastern Spain represent a new lineage within the Bradyrhizobium genus

Lupinus mariae-josephi is a recently described endemic Lupinus species from a small area in Eastern Spain where it thrives in soils with active lime and high pH. The L. mariae-josephi root symbionts were shown to be very slow-growing bacteria with different phenotypic and symbiotic characteristics from those of Bradyrhizobium strains nodulating other Lupinus. Their phylogenetic status was examined by multilocus sequence analyses of four housekeeping genes (16S rRNA, glnII, recA, and atpD) and showed the existence of a distinct evolutionary lineage for L. mariae-josephi that also included Bradyrhizobium jicamae. Within this lineage, the tested isolates clustered in three different sub-groups that might correspond to novel sister Bradyrhizobium species. These core gene analyses consistently showed that all the endosymbiotic bacteria isolated from other Lupinus species of the Iberian Peninsula were related to strains of the B. canariense or B. japonicum lineages and were separate from the L. mariae-josephi isolates. Phylogenetic analysis based on nodC symbiotic gene sequences showed that L. mariae-josephi bacteria also constituted a new symbiotic lineage distant from those previously defined in the genus Bradyrhizobium. In contrast, the nodC genes of isolates from other Lupinus spp. from the Iberian Peninsula were again clearly related to the B. canariense and B. japonicum bv. genistearum lineages. Speciation of L. mariae-josephi bradyrhizobia may result from the colonization of a singular habitat by their unique legume host.

Über die bei der Schwarzerle (Alnus glutinosa) und der gewöhnlichen Garten-Lupine (Lupinus mutabilis) auftretenden Wurzelanschwellungen. Der Akademie vorgelegt am 24. Mai 1866.

Series, title and imprint also in Russian.

Cadmium phytoextraction capacity of white lupine ( Lupinus albus L.) and narrow-leafed lupine ( Lupinus angustifolius L.) in three contrasting agroclimatic conditions of Chile

The phytoextraction process implies the use of plants to promote the elimination of metal contaminants in the soil. In fact, metal-accumulating plants are planted or transplanted in metal-contaminated soil and cultivated in accordance with established agricultural practices. The objective of the present study was to evaluate the productivity and Cd phytoextraction capacity of white lupine ( Lupinus albus L.) and narrow-leafed lupine ( Lupinus angustifolius L.), as well as the effect on residual Cd concentration in the soil. Both species of lupines were grown at three CdCl2 rates (0, 1, and 2 mg kg-1), under three agroclimatic conditions in Chile in 2013. In the arid zone (Pan de Azúcar, 73 mm precipitation), narrow-leafed lupine production was significantly (P < 0.05) higher than white lupine (4.55 vs. 3.26 Mg DM ha-1, respectively). In locations with higher precipitation (Santa Rosa, 670 mm; Carillanca, 880 mm), narrow-leafed lupine DM production was slightly higher than in Pan de Azúcar, but white lupine was approximately three times higher. Total plant Cd concentrations in white and narrow-leafed lupine increased as Cd rates increased in the three environments, but they were much higher in narrow-leafed lupine than white lupine; 150%, 58%, and 344% higher in Pan de Azúcar, Santa Rosa, and Carillanca, respectively. Cadmium uptake (g Cd ha-1) and apparent recovery were also higher (P < 0.05) in narrow-leafed lupine in two environments (Pan de Azúcar and Carillanca). These results suggest that narrow-leafed lupine present higher potential as phytoremediation species than white lupine.

Root effects on the turnover of grain legume residues in soil

Das Ziel dieser Arbeit war, die Einflüsse von Wurzeln und Rhizodeposition auf den Umsatz von Körnerleguminosenresiduen und damit verknüpfte mikrobielle Prozesse zu untersuchen. In einem integrierten Versuch wurden Ackerbohne (Vicia faba L.), Erbse (Pisum sativum L.) und Weiße Lupine (Lupinus albus L.) untersucht. Der Versuch bestand aus drei Teilen, zwei Gefäß-Experimenten und einem Inkubationsexperiment, in denen ausgehend von einem Gefäß-Experiment derselbe Boden und dasselbe Pflanzenmaterial verwendet wurden. In Experiment I wurde die Stickstoff-Rhizodeposition der Körnerleguminosenarten, definiert als wurzelbürtiger N nach dem Entfernen aller sichtbaren Wurzeln im Boden, gemessen und der Verbleib des Rhizodepositions-N in verschiednenen Bodenpools untersucht. Dazu wurden die Leguminosen in einem Gefäßversuch unter Verwendung einer in situ 15N-Docht-Methode mit einer 15N Harnstofflösung pulsmarkiert. In Experiment II wurde der Umsatz der N-Rhizodeposition der Körnerleguminosen und der Einfluss der Rhizodeposition auf den anschließenden C- und N-Umsatz der Körnerleguminosenresiduen in einem Inkubationsexperiment untersucht. In Experiment III wurde der N-Transfer aus den Körnerleguminosenresiduen einschließlich N-Rhizodeposition in die mikrobielle Biomasse und die Folgefrüchte Weizen (Triticum aestivum L.) und Raps (Brassica napus L.) in einem Gewächshaus-Gefäßversuch ermittelt. Die in situ 15N Docht-Markierungs-Methode wies hohe 15N Wiederfindungsraten von ungefähr 84 Prozent für alle drei Leguminosenarten auf und zeigte eine vergleichsweise homogene 15N Verteilung zwischen verschiedenen Pflanzenteilen zur Reife. Die Wurzeln zeigten deutliche Effekte auf die N-Dynamik nach dem Anbau von Körnerleguminosen. Die Effekte konnten auf die N-Rhizodeposition und deren anschließenden Umsatz, Einflüsse der Rhizodeposition von Körnerleguminosen auf den anschließenden Umsatz ihrer Residuen (Stängel, Blätter, erfassbare Wurzeln) und die Wirkungen nachfolgender Nichtleguminosen auf den Umsatzprozess der Residuen zurückgeführt werden: Die N-Rhizodeposition betrug zur Reife der Pflanzen bezogen auf die Gesamt-N- Aufnahme 13 Prozent bei Ackerbohne und Erbse und 16 Prozent bei Weißer Lupine. Bezogen auf den Residual N nach Ernte der Körner erhöhte sich der relative Anteil auf 35 - 44 Prozent. Die N-Rhizodeposition ist daher ein wesentlicher Pool für die N-Bilanz von Körnerleguminosen und trägt wesentlich zur Erklärung positiver Fruchtfolgeeffekte nach Körnerleguminosen bei. 7 - 21 Prozent des Rhizodepositions-N wurden als Feinwurzeln nach Nasssiebung (200 µm) wiedergefunden. Nur 14 - 18 Prozent des Rhizodepositions-N wurde in der mikrobiellen Biomasse und ein sehr kleiner Anteil von 3 - 7 Prozent in der mineralischen N Fraktion gefunden. 48 bis 72 Prozent der N-Rhizodeposition konnte in keinem der untersuchten Pools nachgewiesen werden. Dieser Teil dürfte als mikrobielle Residualmasse immobilisiert worden sein. Nach 168 Tagen Inkubation wurden 21 bis 27 Prozent des Rhizodepositions-N in den mineralisiert. Der mineralisierte N stammte im wesentlichen aus zwei Pools: Zwischen 30 Prozent und 55 Prozent wurde aus der mikrobiellen Residualmasse mineralisiert und eine kleinere Menge stammte aus der mikrobielle Biomasse. Der Einfluss der Rhizodeposition auf den Umsatz der Residuen war indifferent. Durch Rhizodeposition wurde die C Mineralisierung der Leguminosenresiduen nur in der Lupinenvariante erhöht, wobei der mikrobielle N und die Bildung von mikrobieller Residualmasse aus den Leguminosenresiduen in allen Varianten durch Rhizodepositionseinflüsse erhöht waren. Das Potential des residualen Körnerleguminosen-N für die N Ernährung von Folgefrüchten war gering. Nur 8 - 12 Prozent des residualen N wurden in den Folgenfrüchten Weizen und Raps wiedergefunden. Durch die Berücksichtigung des Rhizodepositions-N war der relative Anteil des Residual-N bezogen auf die Gesamt-N-Aufnahme der Folgefrucht hoch und betrug zwischen 18 und 46 Prozent. Dies lässt auf einen höheren N-Beitrag der Körnerleguminosen schließen als bisher angenommen wurde. Die residuale N-Aufnahme von Weizen von der Blüte bis zur Reife wurde durch den Residual-N gespeist, der zur Blüte in der mikrobiellen Biomasse immobilisiert worden war. Die gesamte Poolgröße, Residual-N in der mikrobiellen Biomasse und in Weizen, veränderte sich von der Blüte bis zur Reife nicht. Jedoch konnte ein Rest von 80 Prozent des Residual-N in keinem der untersuchten Pools nachgewiesen werden und dürfte als mikrobielle Residualmasse immobilisiert worden sein oder ist noch nicht abgebaut worden. Die zwei unterschiedlichen Folgefrüchte - Weizen und Raps - zeigten sehr ähnliche Muster bei der N-Aufnahme, der Residual-N Wiederfindung und bei mikrobiellen Parametern für die Residuen der drei Körnerleguminosenarten. Ein differenzierender Effekt auf den Umsatz der Residuen bzw. auf das Residual-N-Aneignungsvermögen der Folgefrüchte konnte nicht beobachtet werden.

Avaliação das silagens de capim-elefante aditivadas com nabo forrageiro, pinhão manso e tremoço, pela técnica de produção de gases

Objetivou-se, na condução deste trabalho, a avaliação das silagens de capim-elefante aditivadas com tortas de nabo forrageiro, pinhão manso e tremoço pela técnica de produção de gás. O experimento foi desenvolvido no Laboratório de Nutrição Animal do Centro de Energia Nuclear na Agricultura da Universidade de São Paulo (LANA/CENA/USP). Como doadores de líquido de rúmen, foram utilizados 2 ovinos da raça Santa Inês, machos, adultos, castrados e providos de cânula ruminal permanente. A alimentação dos animais doadores foi constituída de forragem de gramínea cultivada e uma suplementação, ao final do dia, com feno de Tifton, concentrado comercial e sal mineral à vontade. Os substratos foram secos a 60ºC, moídos em moinho do tipo Willey, provido de peneira com perfurações de 2 mm. Os gases produzidos durante os diferentes períodos de fermentação (0, 4, 8, 12, 24, 36, 48, 72 e 96 h) foram medidos com um transducer - medidor de pressão.O experimento foi instalado segundo um delineamento de blocos ao acaso em que os tratamentos foram arranjados em um esquema de parcelas subdivididas no tempo. Os maiores valores de produção de gás observados para os tratamentos em que adicionou-se torta de tremoço quando comparados com as outras tortas, decorreu do fato da torta de tremoço apresentar menor teor de fibras, propiciando assim, uma maior fermentação ruminal e, consequentemente, maior produção de gás em relação a outros alimentos com maior proporção de carboidratos estruturais (parede celular).As taxas de degradação da fração solúvel da matéria seca foi menor para NF 8% e PM 11% em relação às outras silagens estudadas. Foram encontradas diferenças significativas para as TNF, TPM e TT, nos diferentes níveis, em relação ao volume de gases em 96 h de incubação (P<0,05). As silagens contendo torta de tremoço apresentaram maior produção de gases quando comparadas Às outras tortas. em todos os tratamentos, exceto naqueles em que adicionou-se TT, houve diminuição (P<0,05) nos valores médios de degradabilidade da matéria seca às 96 horas, à medida que aumentou-se o nível de inclusão das tortas. As silagens de capim-elefante adicionadas de tortas de nabo forrageiro ou tremoço, nos diferentes níveis, apresentaram maiores taxas de degradação e maiores produções de gases que as adicionadas de torta de pinhão manso.

Avaliação dos efeitos citotóxicos, genotóxicos, mutagênicos, antigenotóxicos, antimutagênicos e anticarcinogênicos do Lupinus albus em camundongos Mus musculus

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Fracionamento da proteína e estudo termoanalítico das leguminosas: grão de bico (Cicer arietinum), variedade Cíciero e tremoço branco (Lupinus albus L.)

Pós-graduação em Alimentos e Nutrição - FCFAR

Otimização para a obtençao de extrato aquoso de tremoço branco (Lupinus albus L.) adicionado de suco de pitanga

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeito do consumo de tremoço (Lupinus albus) e seu isolado protéico no metabolismo do colesterol em hamsters hipercolesterolemizados

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Desempenho operacional de semeadora em função de mecanismos de corte, velocidade e solos, no sistema plantio direto do milho

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Formulação de extrato aquoso de tremoço branco (Lupinus albus L.) adicionado de suco de pitanga utilizando metodologia de superfície de resposta

The aim of the present study was to verify, through the response surface methodology, the optimum conditions for the combined use of aqueous extract of white lupine (Lupinus albus L.) and tropical pitanga juice (Eugenia unifl ora L.) in the development of a drink. Therefore, the independent variables were represented by the volume of aqueous extract of lupine (mL) and by the volume of pitanga juice (mL). The dependent variables (responses) were obtained through sensory test of acceptance (“appearance”, “aroma”, “taste” and “overall acceptability”). The combined optimization of the variables pointed the second formulation (50mL of aqueous extract of lupine and 30mL of pitanga juice) of the experimental project as the most adequate. In conclusion it was possible to get a drink with attractive sensory characteristics using aqueous extract of white lupine and tropical pitanga juice. The usage of response surface methodology made possible the determination of regions of maximum acceptance for each attribute evaluated in the formulation of a drink with aqueous extract of white lupine and pitanga juice, with a minimum of tests.

In vitro evaluation, in vivo quantification, and microbial diversity studies of nutritional strategies for reducing enteric methane production

The main objective of the present work was to study nutritive strategies for lessening the CH4 formation associated to ruminant tropical diets. In vitro gas production technique was used for evaluating the effect of tannin-rich plants, essential oils, and biodiesel co-products on CH4 formation in three individual studies and a small chamber system to measure CH4 released by sheep for in vivo studies was developed. Microbial rumen population diversity from in vitro assays was studied using qPCR. In vitro studies with tanniniferous plants, herbal plant essential oils derived from thyme, fennel, ginger, black seed, and Eucalyptus oil (EuO) added to the basal diet and cakes of oleaginous plants (cotton, palm, castor plant, turnip, and lupine), which were included in the basal diet to replace soybean meal, presented significant differences regarding fermentation gas production and CH4 formation. In vivo assays were performed according to the results of the in vitro assays. , when supplemented to a basal diet (Tifton-85 hay sp, corn grain, soybean meal, cotton seed meal, and mineral mixture) fed to adult Santa Ines sheep reduced enteric CH4 emission but the supplementation of the basal diet with EuO did not affect ( > 0.05) methane released. Regarding the microbial studies of rumen population diversity using qPCR with DNA samples collected from the in vitro trials, the results showed shifts in microbial communities of the tannin-rich plants in relation to control plant. This research demonstrated that tannin-rich , essential oil from eucalyptus, and biodiesel co-products either in vitro or in vivo assays showed potential to mitigate CH4 emission in ruminants. The microbial community study suggested that the reduction in CH4 production may be attributed to a decrease in fermentable substrate rather than to a direct effect on methanogenesis.

PepT1 mRNA expression levels in sea bream (Sparus aurata) fed different plant protein sources

[EN] The expression and regulation of intestinal oligopeptide transporter (PepT)-1 when vegetable sources are used as a substitute for fish meal in the diet of marine fish has not yet been explored. In the present study, as part of our ongoing work on elucidating PepT1 gene expression in relation to different dietary treatments, we have now isolated and deposited in Genbank database (accession no. GU733710) a cDNA sequence representing the PepT1 in the sea bream (Sparus aurata). The ?de novo? prediction of the three-dimensional structure of PepT1 protein is presented. We also analyzed diet-induced changes in the expression of PepT1 mRNA via real-time RT-PCR using the standard curve method. Sea bream were fed for 140 days with one of the following four diet formulations (43% protein/21% lipid): a control fast growth-promoting diet (C), and three diets with the same formulation but in which 15% of the fish meal was substituted by protein concentrates either from lupine (LPC), chick pea (CPC), or green pea (PPC). Fish fed PPC had significantly (p < 0.05) lower levels of PepT1 transcripts in the proximal intestine than the controls, whereas PepT1 transcript levels in fish fed LPC or CPC were not significantly different from the controls. Although growth was similar between fish fed with different diets during the first 72 days of feeding, growth of the fish fed with PPC was reduced during the second part of the trial and was significantly (p < 0.05) lower than fish fed LPC and CPC diets by the end of the experiment. Correlation between these results and fish growth performances highlights that the intestinal PepT1 mRNA level may serve as a useful marker of the dietary protein quality and absorption efficiency.

Conservation of endangered Lupinus mariae-josephae in its natural habitat by inoculation with selected, native Bradyrhizobium strains

Lupinus mariae-josephae is a recently discovered endemism that is only found in alkaline-limed soils, a unique habitat for lupines, from a small area in Valencia region (Spain). In these soils, L. mariae-josephae grows in just a few defined patches, and previous conservation efforts directed towards controlled plant reproduction have been unsuccessful. We have previously shown that L. mariae-josephae plants establish a specific root nodule symbiosis with bradyrhizobia present in those soils, and we reasoned that the paucity of these bacteria in soils might contribute to the lack of success in reproducing plants for conservation purposes. Greenhouse experiments using L. mariae-josephae trap-plants showed the absence or near absence of L. mariae-josephae-nodulating bacteria in ‘‘terra rossa’’ soils of Valencia outside of L. mariaejosephae plant patches, and in other ‘‘terra rossa’’ or alkaline red soils of the Iberian Peninsula and Balearic Islands outside of the Valencia L. mariae-josephae endemism region. Among the bradyrhizobia able to establish an efficient symbiosis with L. mariae-josephae plants, two strains, LmjC and LmjM3 were selected as inoculum for seed coating. Two planting experiments were carried out in consecutive years under natural conditions in areas with edapho-climatic characteristics identical to those sustaining natural L. mariae-josephae populations, and successful reproduction of the plant was achieved. Interestingly, the successful reproductive cycle was absolutely dependent on seedling inoculation with effective bradyrhizobia, and optimal performance was observed in plants inoculated with LmjC, a strain that had previously shown the most efficient behavior under controlled conditions. Our results define conditions for L. mariae-josephae conservation and for extension to alkaline-limed soil habitats, where no other known lupine can thrive.