Impacts of Natural Selection on Evolution of Core and Symbiotically Specialized (sym) Genes in the Polytypic Species Neorhizobium galegae.

Nodule bacteria (rhizobia) represent a suitable model to address a range of fundamental genetic problems, including the impacts of natural selection on the evolution of symbiotic microorganisms. Rhizobia possess multipartite genomes in which symbiotically specialized (sym) genes differ from core genes in their natural histories. Diversification of sym genes is responsible for rhizobia microevolution, which depends on host-induced natural selection. By contrast, diversification of core genes is responsible for rhizobia speciation, which occurs under the impacts of still unknown selective factors. In this paper, we demonstrate that in goat's rue rhizobia (Neorhizobium galegae) populations collected at North Caucasus, representing two host-specific biovars orientalis and officianalis (N2-fixing symbionts of Galega orientalis and G. officinalis), the evolutionary mechanisms are different for core and sym genes. In both N. galegae biovars, core genes are more polymorphic than sym genes. In bv. orientalis, the evolution of core genes occurs under the impacts of driving selection (dN/dS > 1), while the evolution of sym genes is close to neutral (dN/dS ≈ 1). In bv. officinalis, the evolution of core genes is neutral, while for sym genes, it is dependent on purifying selection (dN/dS < 1). A marked phylogenetic congruence of core and sym genes revealed using ANI analysis may be due to a low intensity of gene transfer within and between N. galegae biovars. Polymorphism in both gene groups and the impacts of driving selection on core gene evolution are more pronounced in bv. orientalis than in bv. officianalis, reflecting the diversities of their respective host plant species. In bv. orientalis, a highly significant (P0 < 0.001) positive correlation is revealed between the p-distance and dN/dS values for core genes, while in bv. officinalis, this correlation is of low significance (0.05 < P0 < 0.10). For sym genes, the correlation between p-distance and dN/dS values is negative in bv. officinalis but is not revealed in bv. orientalis. These data, along with the functional annotation of core genes implemented using Gene Ontology tools, suggest that the evolution of bv. officinalis is based mostly on adaptation for in planta niches while in bv. orientalis, evolution presumably depends on adaptation for soil niches. New insights into the tradeoff between natural selection and genetic diversity are presented, suggesting that gene nucleotide polymorphism may be extended by driving selection only in ecologically versatile organisms capable of supporting a broad spectrum of gene alleles in their gene pools.

Biosynthesized Ag nanoparticles on urea-based periodic mesoporous organosilica enhance galegine content in Galega.

The biological approach for synthesizing nanoparticles (NPs) using plant extracts is an efficient alternative to conventional physicochemical methods. Galegine, isolated from Galega (Galega officinalis L.), has anti-diabetic properties. In the present study, silver nanoparticles (AgNPs) loaded onto urea-based periodic mesoporous organosilica (AgNPs/Ur-PMO) were bio-synthesized using G. officinalis leaf extract. The synthesized NPs were characterized and confirmed via analysis methods. Different concentrations of biosynthesized AgNPs/Ur-PMO nanoparticles (0, 1, 5, 10, and 20 mg L-1) were used as elicitors in cell suspension culture (CSC) of G. officinalis. The callus cells from hypocotyl explants were treated at their logarithmic growth phase (8th d) and were collected at time intervals of 24, 72, 120, and 168 h. The viability and growth of cells were reduced (by 17% and 35%, respectively) at higher concentrations and longer treatments of AgNPs/Ur-PMO; however, the contents of hydrogen peroxide (H2O2) and malondialdehyde (MDA) were increased (1.23 and 3.01 fold, respectively in comparison with the control average). The highest total phenolic (2.43 mg g-1 dry weight) and flavonoid (2.22 mg g-1 dry weight) contents were obtained 168 h after treatment with 10 mg L-1 AgNPs/Ur-PMO. An increasing tendency in the antioxidant enzyme activities was also observed in all the elicitor concentrations. Treatment with AgNPs/Ur-PMO (in particular 5 mg L-1 for 120 h) significantly enhanced the galegine content (up to 17.42 mg g-1) about 1.80 fold compared with the control. The results suggest that AgNPs/Ur-PMO can be used as an effective elicitor for enhancing galegine production in the CSC of G. officinalis. KEY POINTS: • The green biosynthesis of AgNPs/Ur-PMO was done using G. officinalis leaf extract • Its toxicity as an elicitor increased with increasing concentration and treatment time • AgNPs/Ur-PMO significantly increased the antioxidant capacity and galegine content.

Analysis of Complete Chloroplast Genome: Structure, Phylogenetic Relationships of Galega orientalis and Evolutionary Inference of Galegeae.

Galega orientalis, a leguminous herb in the Fabaceae family, is an ecologically and economically important species widely cultivated for its strong stress resistance and high protein content. However, genomic information of Galega orientalis has not been reported, which limiting its evolutionary analysis. The small genome size makes chloroplast relatively easy to obtain genomic sequence for phylogenetic studies and molecular marker development. Here, the chloroplast genome of Galega orientalis was sequenced and annotated. The results showed that the chloroplast genome of G. orientalis is 125,280 bp in length with GC content of 34.11%. A total of 107 genes were identified, including 74 protein-coding genes, 29 tRNAs and four rRNAs. One inverted repeat (IR) region was lost in the chloroplast genome of G. orientalis. In addition, five genes (rpl22, ycf2, rps16, trnE-UUC and pbf1) were lost compared with the chloroplast genome of its related species G. officinalis. A total of 84 long repeats and 68 simple sequence repeats were detected, which could be used as potential markers in the genetic studies of G. orientalis and related species. We found that the Ka/Ks values of three genes petL, rpl20, and ycf4 were higher than one in the pairwise comparation of G. officinalis and other three Galegeae species (Calophaca sinica, Caragana jubata, Caragana korshinskii), which indicated those three genes were under positive selection. A comparative genomic analysis of 15 Galegeae species showed that most conserved non-coding sequence regions and two genic regions (ycf1 and clpP) were highly divergent, which could be used as DNA barcodes for rapid and accurate species identification. Phylogenetic trees constructed based on the ycf1 and clpP genes confirmed the evolutionary relationships among Galegeae species. In addition, among the 15 Galegeae species analyzed, Galega orientalis had a unique 30-bp intron in the ycf1 gene and Tibetia liangshanensis lacked two introns in the clpP gene, which is contrary to existing conclusion that only Glycyrrhiza species in the IR lacking clade (IRLC) lack two introns. In conclusion, for the first time, the complete chloroplast genome of G. orientalis was determined and annotated, which could provide insights into the unsolved evolutionary relationships within the genus Galegeae.

Geographical discrimination of olive oils from Cv. 'Galega Vulgar'.

Olive oils from seven Portuguese regions were selected to study the effect of the geographical origin on the oils' composition. Quality parameters, fatty acids, tocopherols, hydroxytyrosol and tyrosol derivatives, and oxidative stability were evaluated. All olive oils could be classified as extra virgin, and the geographical origin significantly affected the oils chemical composition. Principal component analysis further confirmed the significant impact of the geographical origin on the composition and, indirectly, on stability of the oils, showing that the evaluated parameters could be used as markers for geographical origin identification. Alternatively, Fourier transform infrared spectroscopy was applied, allowing to establish a linear discriminant model that correctly identified the geographical origin of the olive oils with a mean sensitivity of 99 ± 3 % (internal validation), confirming the impact of the oil origin on its characteristics. This finding allowed foreseeing the future application of the spectroscopy approach as a green, fast and non-invasive authentication tool.

Two Cases of Suspected Poisoning With Goat's Rue (Galega officinalis L.) in Horses.

The present report describes two novel cases of suspected intoxication with Galega officinalis in 6- and 21-year-old Arabian mares displaying acute respiratory signs. Both animals showed signs of pulmonary edema at physical examination, with the oldest of the two also manifesting severe dyspnea and foamy nasal discharge. The mares were grazing on the same meadow with hay available ad libitum. Botanical analysis of the latter showed traces of the toxic plant Galega officinalis (L.), which has been daily ingested at a dose of around 14 g of dry matter for three days. Based on the respiratory signs and the presence of goat's rue in the mares' feed, a presumptive diagnosis of plant poisoning was assumed. Dietary change and treatment allowed the 6-year-old mare to fully recover in 3 days while a longer period of about 2 weeks was necessary for the older horse. Horses avoid eating fresh goat's rue as its palatability is low, yet poisoning may still happen in these species when the plant is found in dried and processed feed material.

Investigation of the Phytochemical Composition, Antioxidant Activity, and Methylglyoxal Trapping Effect of Galega officinalis L. Herb In Vitro.

Galega officinalis L. has been known for centuries as an herbal medicine used to alleviate the symptoms of diabetes, but its comprehensive chemical composition and pharmacological activity are still insufficiently known. The current study involved the qualitative and quantitative phytochemical analysis and in vitro evaluation of the antioxidative and methylglyoxal (MGO) trapping properties of galega herb. Ultra high-performance liquid chromatography coupled with both the electrospray ionization mass spectrometer and diode-array detector (UHPLC-ESI-MS and UHPLC-DAD) were used to investigate the composition and evaluate the anti-MGO capability of extracts and their components. Hot water and aqueous methanol extracts, as well as individual compounds representing phytochemical groups, were also assessed for antioxidant activity using DPPH (2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl) and ABTS (2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) assays. Quercetin and metformin were used as a positive control. We confirmed the presence of tricyclic quinazoline alkaloids, guanidines, flavonoids, and hydroxycinnamic acids (HCAs) in galega extracts. The polyphenolic fraction was dominated by mono-, di-, and triglycosylated flavonols, as well as monocaffeoylhexaric acids. The in vitro tests indicated which G. officinalis components exhibit beneficial antioxidative and MGO trapping effects. For galega extracts, flavonols, and HCAs, a potent antiradical activity was observed. The ability to trap MGO was noted for guanidines and flavonoids, whereas HCA esters and quinazoline alkaloids were ineffective. The formation of mono-MGO adducts of galegine, hydroxygalegine, and rutin in the examined water infusion was observed.

Metformin: The Answer to Cancer in a Flower? Current Knowledge and Future Prospects of Metformin as an Anti-Cancer Agent in Breast Cancer.

Interest has grown in studying the possible use of well-known anti-diabetic drugs as anti-cancer agents individually or in combination with, frequently used, chemotherapeutic agents and/or radiation, owing to the fact that diabetes heightens the risk, incidence, and rapid progression of cancers, including breast cancer, in an individual. In this regard, metformin (1, 1-dimethylbiguanide), well known as 'Glucophage' among diabetics, was reported to be cancer preventive while also being a potent anti-proliferative and anti-cancer agent. While meta-analysis studies reported a lower risk and incidence of breast cancer among diabetic individuals on a metformin treatment regimen, several in vitro, pre-clinical, and clinical studies reported the efficacy of using metformin individually as an anti-cancer/anti-tumor agent or in combination with chemotherapeutic drugs or radiation in the treatment of different forms of breast cancer. However, unanswered questions remain with regards to areas such as cancer treatment specific therapeutic dosing of metformin, specificity to cancer cells at high concentrations, resistance to metformin therapy, efficacy of combinatory therapeutic approaches, post-therapeutic relapse of the disease, and efficacy in cancer prevention in non-diabetic individuals. In the current article, we discuss the biology of metformin and its molecular mechanism of action, the existing cellular, pre-clinical, and clinical studies that have tested the anti-tumor potential of metformin as a potential anti-cancer/anti-tumor agent in breast cancer therapy, and outline the future prospects and directions for a better understanding and re-purposing of metformin as an anti-cancer drug in the treatment of breast cancer.

Green synthesis of silver nanoparticles: effect of synthesis reaction parameters on antimicrobial activity.

In this work, the biosynthesis of silver nanoparticles by Galega officinalis extract using AgNO3 as a precursor was reported. The reaction parameters for the biosynthesis and efficiency in their antimicrobial control against Escherichia coli, Staphylococcus aureus and Pseudomonas syringae were determined. For biosynthesis, a central composite design combined with response surface methodology was used to optimize the process parameters (pH, AgNO3 and extract concentration), and the design was assessed through the size distribution, zeta potential and polydispersity index of the nanoparticles. The results demonstrated that at pH 11, 1.6 mM of AgNO3 and 15% vv-1 of G. officinalis extract were the optimal reaction parameters. Transmission electron microscope (TEM) images and X-ray diffraction (XRD) confirmed the formation of small spherical silver nanoparticles. Antimicrobial assays showed a high inhibitory effect against E. coli, S. aureus and P. syringae, and that effect was larger with silver nanoparticles of a smaller size (23 nm). This work demonstrates that G. officinalis extract is a feasible medium for the synthesis of silver nanoparticles and that the control of the reaction parameters can determine the nanoparticle characteristics and therefore their antimicrobial effectiveness.

AOX1-Subfamily Gene Members in Olea europaea cv. "Galega Vulgar"-Gene Characterization and Expression of Transcripts during IBA-Induced in Vitro Adventitious Rooting.

Propagation of some Olea europaea L. cultivars is strongly limited due to recalcitrant behavior in adventitious root formation by semi-hardwood cuttings. One example is the cultivar "Galega vulgar". The formation of adventitious roots is considered a morphological response to stress. Alternative oxidase (AOX) is the terminal oxidase of the alternative pathway of the plant mitochondrial electron transport chain. This enzyme is well known to be induced in response to several biotic and abiotic stress situations. This work aimed to characterize the alternative oxidase 1 (AOX1)-subfamily in olive and to analyze the expression of transcripts during the indole-3-butyric acid (IBA)-induced in vitro adventitious rooting (AR) process. OeAOX1a (acc. no. MF410318) and OeAOX1d (acc. no. MF410319) were identified, as well as different transcript variants for both genes which resulted from alternative polyadenylation events. A correlation between transcript accumulation of both OeAOX1a and OeAOX1d transcripts and the three distinct phases (induction, initiation, and expression) of the AR process in olive was observed. Olive AOX1 genes seem to be associated with the induction and development of adventitious roots in IBA-treated explants. A better understanding of the molecular mechanisms underlying the stimulus needed for the induction of adventitious roots may help to develop more targeted and effective rooting induction protocols in order to improve the rooting ability of difficult-to-root cultivars.

Galega officinalis extract regulate the diabetes mellitus related violations of proliferation, functions and apoptosis of leukocytes.

BACKGROUND: An impaired leukocytes function is the factor causing the susceptibility of patients with diabetes mellitus to infections. The outmost importance for the understanding of the immunological processes involved in diabetes pathogenesis is to give the characteritics of the immunological profile and changes therein, during the course of desease. Long-used in folk medicine to treat diabetes Galega officinalis L. has been chosen for the correction of the immune system dysfunction. METHODS: The experiments were conducted on male Wistar rats. Fractionation of bone marrow cells suspension was performed in a three-layer ficoll-sodium amidotrizoate density gradient. The lymphocytic-granulocytic cells proliferative activity was studied using enzyme immunoassay with 5-bromo-2'-deoxyuridine (BrdU). For staining of bone marrow preparations May-Gruenwald-Romanowsky-Giemsa (Pappenheim) method was used. To evaluate the content of cationic proteins and myeloperoxidase in neutrophilic leukocytes cytochemical studies were performed. Content of tumor necrosis factor alpha was carried out by immuno-enzymatic analysis. Lymphocytes apoptosis was examined by fluorescent analysis using annexin V. RESULTS: Diabetes mellitus development was accompanied with violation of neutrophils and lymphocytes proliferation, increased activity of myeloperoxidase and enhanced apoptosis process. Administration of Galega officinalis extract under the condition of diabetes promotes the restoration of neutrophils bone marrow pool and the reduction of lymphoblasts number and causes inhibition of the lymphocytes apoptosis process. CONCLUSIONS: Investigated medicine has a pronounced immunocorrective effect under the conditions of diabetes mellitus and can become the basis for creating a new generation of antidiabetic drugs.

The Long-Term Efficacy of a Galactagogue Containing Sylimarin-Phosphatidylserine and Galega on Milk Production of Mothers of Preterm Infants.

AIMS: To investigate the efficacy of a galactagogue, containing Sylimarin-phosphatidylserine (SILITIDIL) and galega consumed in the first month after delivery by mothers of preterm infants, in maintaining milk production during the first 3-6 months after delivery. MATERIALS AND METHODS: Mothers of infants born at gestational age (GA) between 27 and 32 weeks, enrolled in our previous prospective, double-blind, randomized trial and randomly allocated to receive either the galactagogue (GG) or a placebo (PG), were asked about their milk production at 3 and 6 months after delivery. RESULTS: Of the 100 mothers involved in this study, 45 of GG and 44 of PG responded comprehensively to the questions asked. At the third month after delivery, exclusive human milk administration was reported by 22 mothers of GG and 12 mothers of PG (p < 0.05), whereas 29 mothers of GG and 18 mothers of PG were able to administer >50% of the amount of milk assumed. At the sixth month of life, only eight infants received exclusive human milk (six and two of GG and PG, respectively), and the data are not sufficient for a meaningful clinical evaluation. CONCLUSIONS: It is assumed that a galactagogue during the first month after delivery improves human milk administration to preterm neonates after discharge and for the first 3 months of life.

The role of natural galactagogues during breast feeding: focus on a Galega officinalis based food supplement.

Maternal milk is the optimal food for newborns. To this end, a number of interventions are used to enhance milk production. However, pharmacological interventions may be associated with a perceived risk of adverse effects and therefore many mothers prefer to rely on natural herbal remedies. Several herbal remedies have been traditionally used to this purpose. However, the level of evidence supporting their use is mixed. Among different currently-employed natural remedies, Galega officinalis has emerged to be one of those sustained by the strongest evidence. In this paper, we comment on a galega-based product. It is a standardized food supplement used to support breastfeeding mothers and to promote milk production containing Galega officinalis and other substances, including vitamins and magnesium, with potential effect on mother's well-being. In a recent product evaluation on a large sample of Italian women, the wide majority of mothers have declared to be satisfied with this product, and two third of them reported that the milk production was improved with the use of this product. Noteworthy, this galega-based food supplement was also reported to promote psychological benefit. The evidence of a perceived psychological benefit associated with this product is of particular importance, given the high degree of distress often experienced by mothers during the post-partum period. Last, a high level of safety was perceived by the participants. This galega-based food supplement does have a role in supporting breastfeeding mothers and enhance milk production during lactation. Further clinical trials could provide further evidence on the effectiveness of the product.

Efficacy of a galactogogue containing silymarin-phosphatidylserine and galega in mothers of preterm infants: a randomized controlled trial.

BACKGROUND/OBJECTIVES: Human milk (HM) is the best possible food for all infants, especially for preterm ones, but lactation and breastfeeding are very difficult for mothers of preterm babies and high rates of breastfeeding difficulties have been reported. Our aim was to investigate the efficacy of a galactogogue containing silymarin-phosphatidylserine and galega in increasing milk production during the first month after delivery in a population of mothers of preterm infants. SUBJECTS/METHODS: Mothers of infants with gestational age (GA) between 27+0 and 32+6 weeks were enrolled in this prospective, double-blind, randomized trial and were randomly allocated to receive either the galactogogue containing silymarin-phosphatidylserine and galega, 5 g/day (galactogogue group, GG), or a placebo, 5 g of lactose per day (placebo group, PG) from the 3rd to the 28th day after delivery. RESULTS: Fifty mothers were included in each group. General characteristics of mothers and pregnancies were similar. Milk production was significantly greater in the GG at the 7th day of life and at the 30th day of life. Daily milk production from the 7th to the 30th day of life was 200 (110-380) ml in the GG vs 115 (60-245) ml in the PG (P<0.0001). The total production of milk during the study period was significantly higher in the GG (6523±5298 ml vs 4136±4093 ml; P<0.02). At the end of the study, 45 mothers of the GG were able to reach the target of milk supply of 200 ml/day compared with 25 mothers of the PG (P<0.01). No adverse reactions were noticed in the study groups. CONCLUSIONS: Silymarin-phosphatidylserine and galega increased milk production in mothers of preterm infants without any significant side effects.

Productos naturales para el tratamiento de la diabetes (I): mecanismos de acción / Produtos naturais para o tratamento da diabetes (I): mecanismos de acçao / Natural products for the treatment of diabetes (I): mechanism of action

La diabetes mellitus tipo 2 (DMT2) es una enfermedad metabólica caracterizada por un hipergluciemia persistente, la cual, si no es tratada adecuadamente, a largo plazo puede producir complicaciones cardiovasculares, trastornos renales o retinopatía. El desarrollo de la enfermedad puede prevenirse o retrasarse en personas con tolerancia a la glucosa alterada, mediante la implementación de los cambios de estilo de vida o el uso de agentes terapéuticos. Algunos de estos medicamentos se han obtenido a partir de plantas o tienen origen microbiano, como la galegina aislada de Galega officinalis, que tiene una gran similitud estructural con la metformina. Picnogenol, miglitol, acarbosa y voglibosa son otros antidiabéticos de origen natural. En la presente revisión se recopilan los principales artículos sobre plantas medicinales y productos naturales utilizados para el tratamiento de la DMT2 y sus comorbilidades, sobre la base de sus mecanismos de acción como agentes antidiabéticos. S excluyen las drogas vegetales ricas en polisacáridos. La inhibición de la alpha-glucosidasa y la alpha-amilasa, efectos sobre la capacitación y transportadores de glucosa, y la modificación de mecanismos mediados por el receptor activados por proliferadores de peroxisomas (PPAR), la inhibición de la actividad tirosina-fosfatasa 1B (PTP1B), la modificación de la expresión génica y las actividades de hormonas implicadas en la homeostasis de la glucosa, tales como la adiponectina, resistina e incretina, yl a reducción del estrés oxidativo, son algunos de los mecanismos en los que productos naturales están involucrados (AU)

Adiabetes mellitus tipo 2 (DMT2) è uma doença metabólica caracterizada por hiperglicemia persistente, a qual, se nao for adequadamente tratada, a longo prazo pode originar complicaçoes cardiovasculares, disturbios renais ou retinopatía. O desnvolvimiento da doença pode ser prevenido ou adiado empessoas com tolerancia à glicose alterada, através da implementaçao de mudanças no estilo de vido ou do uso de agentes terapéuticos. Alguns destes medicamentos tem origen microbiana ou sao derivados de plantas, tal como a galefina, composto isolado da espècie Galega officinalis, que tem uma grande semelhança estrutural com a metformina. Picnogenol, acarbose, miglitol e Voglibose sao exemplos de outros antiabéticos de origen natural. Esta revisao compila os principais artigos sobre plantas medicinais e productos naturais usados para o tratamento da DMT2 e suas comorbilidades, tendo como base os respectivos mecanismos de acçao como agentes antidiabéticos. As drogas vegetais ricas em policárideos sao excluidas. A inhibiçao de alpha-glicosade e da alpha-amilase, efeitos sobre a captaçao e transportadores de glicose, modificaçao de mecanismos mediados pelos recptores activados por proliferadores de peroxisomas (PPAR), inhibiçao da actividade da tirosina fosfatase 1B (PTB1B), modificaçao da expressao génica e da actividade de hormonas envolvidas na homeostase da glicose, como adiponectina, resistina e incretinas, bem como a reduçao do stress oxidativo, sao alguns dos mecanismos em que os productos naturais estao envueltos (AU)

Type 2 diabetes mellitus is a metabolic disorder characterized by persistent hyperglycemia, which can cause long-term cardiovascular complications, kidney disorders, retinopathy and circulatory problems. Development of disease can be prevented or delayed in people with impaired glucose tolerance, by implementing changes in lifestyle or with the use of therapeutic agents. Some of these medicines are derived from plants or microbial origin, such as galegine isolated from Galega officinalis, which has a great similarity with the antidiabetic drug metformin. Pycnogenol, acarbose, miglitol and voglibose are other antidiabetic agents from natural origin. This review compiles the main articles on medicinal plants and natural products used for the treatment of diabetes and its comorbidities, focusing on their mechanisms of action as antidiabetic agents. Polysacharide containing hebal drugs are excluded. The inhibition of alpha-glucosidase and alpha-amylase, the effect on glucose uptake and glucose transporters, the modification of perosixome proliferator activated receptors (PPAR), inhibition of protein-tyrosine phosphatase activity 1B (PTP1B), modification of gene expression and activities of hormones involved in glucose homeostasis, such as adiponectin, resistin and incretin, as well as the reduction of oxidative stress are some of the mechanisms in which natural products are involved (AU)

Therapeutic Potential of Temperate Forage Legumes: A Review.

The discovery of bioactive molecules from botanical sources is an expanding field, preferentially oriented to plants having a tradition of use in medicine and providing high yields and availability. Temperate forage legumes are Fabaceae species that include worldwide-important crops. These plants possess therapeutic virtues that have not only been used in veterinary and folk medicine, but have also attracted the interest of official medicine. We have examined here Medicago sativa (alfalfa), Trifolium pratense and T. repens (clovers), Melilotus albus and M. officinalis (sweet clovers), Lotus corniculatus (birdsfoot trefoil), Onobrychis viciifolia (sainfoin), Lespedeza capitata (roundhead lespedeza), and Galega officinalis (goat's rue). The phytochemical complexes of these species contain secondary metabolites whose pharmacological potentials deserve investigation. Major classes of compounds include alkaloids and amines, cyanogenic glycosides, flavonoids, coumarins, condensed tannins, and saponins. Some of these phytochemicals have been related to antihypercholesterolemia, antidiabetic, antimenopause, anti-inflammatory, antiedema, anthelmintic, and kidney protective effects. Two widely prescribed drugs have been developed starting from temperate forage legumes, namely, the antithrombotic warfarin, inspired from sweet clover's coumarin, and the antidiabetic metformin, a derivative of sainfoin's guanidine. Available evidence suggests that temperate forage legumes are a potentially important resource for the extraction of active principles to be used as nutraceuticals and pharmaceuticals.

[THE ALKALOID-FREE FRACTION FROM Galega officinalis EXTRACT PREVENTS OXIDATIVE STRESS UNDER EXPERIMENTAL DIABETES MELLITUS].

The effect of alkaloid-free fraction from Galega officinalis extract on the process of formation of reactive oxygen species and indicators of prooxidant-antioxidant balance was investigated in rat peripheral blood under conditions of experimental diabetes mellitus. It was shown that alkaloid-free fraction from Galega officinalis extract prevents oxidative stress development in rats with streptozotocin-induced diabetes, providing antioxidant and antiradical mobilization mechanisms to protect the blood system. In the case of extract application to animals with studied pathology, one can observe a reducing effect of reactive oxygen species generation in leukocytes, inhibition of proteins and lipids oxidative modification processes and increased activity of key enzymes of rat peripheral blood antioxidant system (superoxide dismutase, catalase and glutathione peroxidase). The revealed biological effect could be explained by the presence of biologically active substances with antioxidant properties in the extract composition (phytol and flavonoids).

Genomic features separating ten strains of Neorhizobium galegae with different symbiotic phenotypes.

BACKGROUND: The symbiotic phenotype of Neorhizobium galegae, with strains specifically fixing nitrogen with either Galega orientalis or G. officinalis, has made it a target in research on determinants of host specificity in nitrogen fixation. The genomic differences between representative strains of the two symbiovars are, however, relatively small. This introduced a need for a dataset representing a larger bacterial population in order to make better conclusions on characteristics typical for a subset of the species. In this study, we produced draft genomes of eight strains of N. galegae having different symbiotic phenotypes, both with regard to host specificity and nitrogen fixation efficiency. These genomes were analysed together with the previously published complete genomes of N. galegae strains HAMBI 540T and HAMBI 1141. RESULTS: The results showed that the presence of an additional rpoN sigma factor gene in the symbiosis gene region is a characteristic specific to symbiovar orientalis, required for nitrogen fixation. Also the nifQ gene was shown to be crucial for functional symbiosis in both symbiovars. Genome-wide analyses identified additional genes characteristic of strains of the same symbiovar and of strains having similar plant growth promoting properties on Galega orientalis. Many of these genes are involved in transcriptional regulation or in metabolic functions. CONCLUSIONS: The results of this study confirm that the only symbiosis-related gene that is present in one symbiovar of N. galegae but not in the other is an rpoN gene. The specific function of this gene remains to be determined, however. New genes that were identified as specific for strains of one symbiovar may be involved in determining host specificity, while others are defined as potential determinant genes for differences in efficiency of nitrogen fixation.

An aquaporin protein is associated with drought stress tolerance.

Water channel proteins known as aquaporins (AQPs) regulate the movement of water and other small molecules across plant vacuolar and plasma membranes; they are associated with plant tolerance of biotic and abiotic stresses. In this study, a PIP type AQPs gene, designated as GoPIP1, was cloned from Galega orientalis, a high value leguminous forage crop. The GoPIP1 gene consists of an 870 bp open reading frame encoding a protein of 289 amino acids, and belongs to the PIP1 subgroup of the PIP subfamily. The transcript level of GoPIP1 was higher in the root of G. orientalis than in the leaf and stem. The level of GoPIP1 transcript increased significantly when treated with 200 mM NaCl or 20% polyethylene glycol (PEG) 6000. Transient expression of GoPIP1 in onion epidermal cells revealed that the GoPIP1 protein was localized to the plasma membrane. Over-expression of GoPIP1 increased the rosette/root ratio and increased sensitivity to drought in transgenic Arabidopsis plants. However, GoPIP1 over-expression in Arabidopsis had no significant effect under saline condition. The present data provides a gene resource that contributes to furthering our understanding of water channel protein and their application in plant stress tolerance.

[Silymarin/galega administration in term and preterm mothers to sustain breast feeding: an observational study]. / La somministrazione di silimarina/galega per il sostegno dell'allatamento al sone: studio prospettico osservazionale in madri di neonati a termine e pretermine.

AIM: Breastfeeding is considered the most important source of nutrition both in the first year of life for term and preterm infants, because of its well-known positive effects on short and long-term outcome. Unfortunately not all the mothers can begin or maintain an adequate milk secretion. Premature delivery and prolonged length of stay in the hospital can influence maternal milk production due to maternal anxiety and/ or neonatal pathologies related to prematurity. A variety of herbal and pharmaceutical products have been recommended as galactogogues, substances that promote lactation. METHODS: In this observational study, the authors compared the effect on breast milk production of an oral maternal supplementation of two galactogogues (sylimarine and galega-Lutein latte®) in a term and preterm mothers group (preterm infants recovered in neonatal intensive care unit-NICU). RESULTS: The breast milk production significantly reduced at start in preterm mothers (N.=16) versus term mothers (N.=16) (P<0.08), after sylimarine and galega supplementation was the same in both groups within two months of life (P=NS). CONCLUSION: This galactogogues supplementation could be recommended to maintain an adequate lactation in premature mothers, especially when their infants are still recovered in the NICU.

Overexpression of the Galega orientalis gibberellin receptor improves biomass production in transgenic tobacco.

Gibberellins (GAs) are well-known phytohormones that contribute to a wide range of plant growth and development functions including stem elongation and leaf expansion. GA receptors perceive GA and transmit signals to activate GA-regulated reactions. In this study, a GA receptor gene with homology to other leguminous plants was isolated from Galega orientalis and termed GoGID. The 1732-bp full-length GoGID gene included an open reading frame of 1035 bp encoding a peptide of 344 amino acids. Sequence analysis indicated that GoGID shares conserved HGGS motif and active amino acid sites (Ser-Asp-Val/IIe) that are essential for maintaining it GA-binding activity. GoGID mRNA expression was more abundant in leaves than in roots or stems and could be up-regulated by the exogenous hormones. Overexpression of GoGID in transgenic tobacco plants promoted plant elongation and improved biomass production. These results suggested that GoGID functions as a GA receptor to alter GA-mediated signaling. GoGID may have a role in genetic engineering for the improvement of forage crops.