Influence of different Sinorhizobium meliloti inocula on abundance of genes involved in nitrogen transformations in the rhizosphere of alfalfa (Medicago sativa L.)

Inoculation of leguminous seeds with selected rhizobial strains is practised in agriculture to ameliorate the plant yield by enhanced root nodulation and nitrogen uptake of the plant. However, effective symbiosis between legumes and rhizobia does not only depend on the capacity of nitrogen fixation but also on the entire nitrogen turnover in the rhizosphere. We investigated the influence of seed inoculation with two indigenous Sinorhizobium meliloti strains exhibiting different efficiency concerning plant growth promotion on nitrogen turnover processes in the rhizosphere during the growth of alfalfa. Quantification of six target genes (bacterial amoA, nirK, nirS, nosZ, nifH and archaeal amoA) within the nitrogen cycle was performed in rhizosphere samples before nodule formation, at bud development and at the late flowering stage. The results clearly demonstrated that effectiveness of rhizobial inocula is related to abundance of nifH genes in the late flowering phase of alfalfa. Moreover, other genes involved in nitrogen turnover had been affected by the inocula, e.g. higher numbers of amoA copies were observed during flowering when the more effective strain had been inoculated. However, the respective gene abundances differed overall to a greater extent between the three plant development stages than between the inoculation variants.     

Traditional food and herbal uses of wild plants in the ancient South-Slavic diaspora of Mundimitar/Montemitro (Southern Italy)

ABSTRACT: BACKGROUND: In Europe, only a limited number of cross-cultural comparative field studies or meta-analyses have been focused on the dynamics through which folk plant knowledge changes over space and time, while a few studies have contributed to the understanding of how plant uses change among newcomers. Nevertheless, ethnic minority groups and/or linguistic "isles" in Southern and Eastern Europe may provide wonderful arenas for understanding the various factors that influence changes in plant uses. METHODS: A field ethnobotanical study was carried out in Mundimitar (Montemitro in Italian), a village of approx. 450 inhabitants, located in the Molise region of South-Eastern Italy. Mundimitar is a South-Slavic community, composed of the descendants of people who migrated to the area during the first half of the 14th century, probably from the lower Neretva valley (Dalmatia and Herzegovina regions). Eighteen key informants (average age: 63.7) were selected using the snowball sampling technique and participated in in-depth interviews regarding their Traditional Knowledge (TK) of the local flora. RESULTS: Although TK on wild plants is eroded in Montemitro among the youngest generations, fiftyseven taxa (including two cultivated species, which were included due to their unusual uses) were quoted by the study participants. Half of the taxa have correspondence in the Croatian and Herzegovinian folk botanical nomenclature, and the other half with South-Italian folk plant names. A remarkable link to the wild vegetable uses recorded in Dalmatia is evident. A comparison of the collected data with the previous ethnobotanical data of the Molise region and of the entire Italian Peninsula pointed out a few uses that have not been recorded in Italy thus far: the culinary use of boiled black bryony (Tamus communis) shoots in sauces and also on pasta; the use of squirting cucumber (Ecballium elaterium) juice for treating malaria in humans; the aerial parts of the elderberry tree (Sambucus nigra) for treating erysipelas in pigs; the aerial parts of pellitory (Parietaria judaica) in decoctions for treating haemorrhoids. CONCLUSIONS: The fact that half of the most salient species documented in our case study - widely available both in Molise and in Dalmatia and Herzegovina - retain a Slavic name could indicate that they may have also been used in Dalmatia and Herzegovina before the migration took place. However, given the occurrence of several South-Italian plant names and uses, also a remarkable acculturation process affected the Slavic community of Montemitro during these last centuries. Future directions of research should try to simultaneously compare current ethnobotanical knowledge of both migrated communities and their counterparts in the areas of origin.     

Synthesis of macrolones with central piperazine ring in the linker and its influence on antibacterial activity

Three macrolides, clarithromycin, azithromycin and 11-O-Me-azithromycin have been selected for the construction of a series of new macrolone derivatives. Quinolone-linker intermediates are prepared by Sonogashira-type C(6)-alkynylation of 6-iodoquinolone precursors. The final macrolones, differing by macrolide moiety and substituents at the position N-1 of the quinolone or by the presence of an ethyl ester or free acid on the quinolone unit attached via a linker. The linker comprises of a central piperazine ring bonded to the 4″-O position of cladinose by 3-carbon ester or ether functionality. Modifications of the linker did not improve antibacterial properties compared to the previously reported macrolone compounds. Linker flexibility seems to play an important role for potency against macrolide resistant respiratory pathogens.     

Biopsy coupled to quantitative immunofluorescence: a new method to study the human vascular endothelium.

Limited availability of endothelial tissue is a major constraint when investigating the cellular mechanisms of endothelial dysfunction in patients with metabolic and cardiovascular diseases. We propose a novel approach that combines collection of 200-1,000 endothelial cells from a superficial forearm vein or the radial artery, with reliable measurements of protein expression by quantitative immunofluorescence analysis. This method was validated against immunoblot analysis in cultured endothelial cells. Levels of vascular endothelial cell activation, oxidative stress, and nitric oxide synthase expression were measured and compared in five patients with severe chronic heart failure and in four healthy age-matched subjects. In summary, vascular endothelial biopsy coupled with measurement of protein expression by quantitative immunofluorescence analysis provides a novel approach to the study of the vascular endothelium in humans.     

Mapping DNA damage‐dependent genetic interactions in yeast via party mating and barcode fusion genetics

Condition‐dependent genetic interactions can reveal functional relationships between genes that are not evident under standard culture conditions. State‐of‐the‐art yeast genetic interaction mapping, which relies on robotic manipulation of arrays of double‐mutant strains, does not scale readily to multi‐condition studies. Here, we describe barcode fusion genetics to map genetic interactions (BFG‐GI), by which double‐mutant strains generated via en masse “party” mating can also be monitored en masse for growth to detect genetic interactions. By using site‐specific recombination to fuse two DNA barcodes, each representing a specific gene deletion, BFG‐GI enables multiplexed quantitative tracking of double mutants via next‐generation sequencing. We applied BFG‐GI to a matrix of DNA repair genes under nine different conditions, including methyl methanesulfonate (MMS), 4‐nitroquinoline 1‐oxide (4NQO), bleomycin, zeocin, and three other DNA‐damaging environments. BFG‐GI recapitulated known genetic interactions and yielded new condition‐dependent genetic interactions. We validated and further explored a subnetwork of condition‐dependent genetic interactions involving MAG1, SLX4, and genes encoding the Shu complex, and inferred that loss of the Shu complex leads to an increase in the activation of the checkpoint protein kinase Rad53.     

An ethnobotany of the Lukomir Highlanders of Bosnia & Herzegovina

Background

This aim of this study is to report upon traditional knowledge and use of wild medicinal plants by the Highlanders of Lukomir, Bjela?nica, Bosnia and Herzegovina (B&H). The Highlanders are an indigenous community of approximately 60 transhumant pastoralist families who speak Bosnian (Bosanski) and inhabit a highly biodiverse region of Europe. This paper adds to the growing record of traditional use of wild plants within isolated communities in the Balkans.

Methods

An ethnobotanical study using consensus methodology was conducted in Lukomir in Bjela?nica’s mountains and canyons. Field work involved individual semi-structured interviews during which informants described plants, natural product remedies, and preparation methods on field trips, garden tours, while shepherding, or in settings of their choice. Plant use categories were ranked with informant consensus factor and incorporated into a phylogenetic tree. Plants cited were compared to other ethnobotanical surveys of the country.

Results

Twenty five people were interviewed, resulting in identification of 58 species (including two subspecies) from 35 families, which were cited in 307 medicinal, 40 food, and seven material use reports. Individual plant uses had an average consensus of five and a maximum consensus of 15 out of 25. There were a number of rare and endangered species used as poisons or medicine that are endemic to Flora Europaea and found in Lukomir. Ten species (including subspecies) cited in our research have not previously been reported in the systematic ethnobotanical surveys of medicinal plant use in B&H: (Elymus repens (L.) Gould, Euphorbia myrsinites L., Jovibarba hirta (L.) Opiz, Lilium bosniacum (Beck) Fritsch, Matricaria matricarioides (Less.) Porter ex Britton, Phyllitis scolopendrium (L.) Newman, Rubus saxatilis L., Silene uniflora Roth ssp. glareosa (Jord.) Chater & Walters, Silene uniflora Roth ssp. prostrata (Gaudin) Chater & Walters, Smyrnium perfoliatum L.). New uses not reported in any of the aforementioned systematic surveys were cited for a total of 28 species. Thirteen percent of medicinal plants cited are endemic: Helleborus odorus Waldst. et Kit., Gentiana lutea L., Lilium bosniacum (Beck) Fritsch, Silene uniflora Roth ssp. glareosa (Jord.) Chater & Walters., Silene uniflora Roth ssp. prostrata (Gaudin) Chater & Walters, Salvia officinalis L., Jovibarba hirta (L.) Opiz, and Satureja montana L.

Conclusions

These results report on the cohesive tradition of medicinal plant use among healers in Lukomir, Bosnia and Herzegovina. This work facilitates the community’s development by facilitating local and international conversations about their traditional medicine and sharing insight for conservation in one of Europe’s most diverse endemic floristic regions, stewarded by one of Europe’s last traditional Highland peoples.

     

The ecological aspect of ethnobotany and ethnopharmacology of population in Bosnia and Herzegovina

This paper contains first systematical revision of the results on traditional use of wild medicinal and aromatic herbs on the territory of Bosnia and Herzegovina (B&H)--west of Balkan Peninsula; Southeast of Europe. There have been detected 227 plants belonging to 71 different plant families, which are being used with ethno therapeutic purpose. Results were obtained by method of open ethno botanical interview which comprised 150 persons, whose average age was 63. Medicinal plants in ethno therapy are being used either in fresh, raw or dried condition. Different herbal parts, depending on period of vegetation season, sometimes even in winter, are basis for preparation of infusions (59%), decoct (19%), tinctures (4%). Especially original are balms known as Bosnian "mehlems", which are fresh cuted herbal parts mixed with lukewarm resin, raw cow butter or honey. In ethno therapy are mostly being used aerial plant organs. Majority of herbs is being used for treatment of illnesses of respiratory (22%), gastrointestinal (19%) and urinary and genital system (9%), for treatment of skin conditions (11%), as well as for nervous system and heart diseases (16%). The most original plants on the field of ethno pharmacology, comparing with ethno therapy practice of other regions, are as follows: Ballota nigra, Aesculus hippocastanum, Calluna vulgaris, Centaurea cyanus, Euphrasia rostkoviana, Geranium robertianum, Gentiana asclepiadea, Helichrysum italicum, Lycopodium clavatum, Marrubium vulgare, Nepeta cataria, Populus tremula, Ruta graveolens, Tamus communis, Teucrium montanum, T. chamaedrys, and endemic plants Gentiana lutea subsp. symphyandra, Teucrium arduini, Micromeria thymifolia, Satureja montana, S. subspicata, Rhamnus fallax and Viola elegantula. There haven't been noticed significant differences in the frequencies of medicinal plants use among different ethnical groups. But, it has been perceived that longer ethno therapeutic tradition possess inhabitants of sub- and Mediterranean areas, as well as inhabitants of the mountain areas of B&H, regardless their ethnicity.     

Genomic fingerprinting of Bradyrhizobium japonicum isolates by RAPD and rep-PCR

Genetic diversity of indigenous Bradyrhizobium japonicum population in Croatia was studied by using different PCR-based fingerprinting methods. Characteristic DNA profiles for 20 B. japonicum field isolates and two reference strains were obtained using random primers (RAPD) and two sets of repetitive primers (REP- and ERIC-PCR). In comparison with the REP, the ERIC primer set generates fingerprints of lower complexity, but still several strain-specific bands were detected. Different B. japonicum isolates could be more efficiently distinguished by using combined results from REP- and ERIC-PCR. The most polymorphic bands were observed after amplification with four different RAPD primers. Both methods, RAPD and rep-PCR, resulted in identical grouping of the strains. Cluster analysis, irrespective of the fingerprinting method used, revealed that all the isolates could be divided into three major groups. Within the major groups, the degree of relative similarity between B. japonicum isolates was dependent upon the method used. Our results indicate that both RAPD and rep-PCR fingerprinting can effectively distinguish different B. japonicum strains. RAPD fingerprinting proved to be slightly more discriminatory than rep-PCR.     

In-situ phytoextraction of Ni by a native population of Alyssum murale on an ultramafic site (Albania)

Ultramafic outcrops are widespread in Albania and host several Ni hyperaccumulators (e.g., Alyssum murale Waldst. &; Kit.). A field experiment was conducted in Pojske (Eastern Albania), a large ultramafic area in which native A. murale was cultivated. The experiment consisted in testing the phytoextraction potential of already installed natural vegetation (including A. murale) on crop fields with or without suitable fertilisation. The area was divided into six 36-m² plots, three of which were fertilised in April 2005 with (NPK + S). The soil (Magnesic Hypereutric Vertisol) was fully described as well as the mineralogy of horizons and the localisation of Ni bearing phases (TEM-EDX and XRD). Ni availability was also characterised by Isotopic Exchange Kinetics (IEK). The flora was fully described on both fertilised and unfertilised plots and the plant composition (major and trace elements) and biomass (shoots) harvested individually were recorded. The soil had mainly two Ni-bearing phases: high-Mg smectite (1.3% Ni) and serpentine (0.7% Ni), the first one being the source of available Ni. Ni availability was extremely high according to IEK and confirmed by Ni contents in Trifolium nigriscens Viv. reaching 1,442 mg kg^?1 (A new hyperaccumulator?). Total biomass yields were 6.3 t ha^?1 in fertilised plots and 3.2 t ha^?1 in unfertilised plots with a highly significant effect: fertilisation increased dramatically the proportion of A. murale in the plots (2.6 t ha^?1 vs. 0.2 t ha^?1). Ni content in the shoots of A. murale reached 9,129 mg kg^?1 but metal concentration was not significantly affected by fertilisation. Phytoextracted Ni in total harvest reached 25 kg Ni ha^?1 on the fertilised plots. It was significantly lower in unfertilised plots (3 kg Ni ha^?1). Extensive phytomining on such sites could be promising in the Albanian context by domesticating already installed natural populations with fertilisation.