Comparative analysis of prokaryotic diversity in solar salterns in eastern Anatolia (Turkey)

The prokaryotic communities of four salterns (Bingöl, Fadlum, Kemah, and Tuzlagözü) in Turkey were examined and compared using the cultivation and cultivation-independent methods [fluorescence in situ hybridization (FISH) and 454 pyrosequencing]. FISH analysis with universal probes revealed that feeding waters carried 1.6 × 10²–1.7 × 10³ cells mL^?1, while crystallization ponds carried 3.8 × 10⁶–2.0 × 10⁷ cells mL^?1 that were mostly haloarchaea, including square cells (except for Kemah). High-throughput 16S rRNA-based gene sequencing showed that the most frequent archaeal OTUs in Bingöl, Fadlum, Tuzlagözü, and Kemah samples were affiliated with Haloquadratum (76.8 %), Haloarcula (27.8 %), Halorubrum (49.6 %), and Halonotius (59.8 %), respectively. Bacteroidetes was the dominant bacterial phylum in Bingöl and Fadlum, representing 71.5 and 79.5 % of the bacterial OTUs (respectively), while the most abundant bacterial phylum found in the Kemah saltern was Proteobacteria (79.6 %). The majority of the bacterial OTUs recovered from Tuzlagözü belonged to the Cyanobacteria (35.7 %), Bacteroidetes (35.0 %), and Proteobacteria (25.5 %) phyla. Cultivation studies revealed that the archaeal isolates were closely related to the genera Halobacterium, Haloarcula, and Halorubrum. Bacterial isolates were confined to two phyla, Proteobacteria (Alphaproteobacteria and Gammaproteobacteria classes) and Bacteroidetes. Comparative analysis showed that members of the Euryarchaeota, Bacteroidetes, Proteobacteria, and Cyanobacteria phyla were major inhabitants of the solar salterns.     

Non-specific symbiotic germination of <Emphasis Type="Italic">Cynorkis purpurea</Emphasis> (Thouars) Kraezl., a habitat-specific terrestrial orchid from the Central Highlands of Madagascar

Orchids, particularly terrestrial taxa, rely mostly on basidiomycete fungi in the Cantharellales and Sebacinales that trigger the process of seed germination and/or initiate the full development of the seedling. During the course of development, orchids may associate with the same fungus, or they may enlist other types of fungi for their developmental needs leading to resilience in a natural setting. This study examined in vitro seed germination and seedling developmental behavior of Cynorkis purpurea, a terrestrial orchid from the Central Highlands of Madagascar. This species is mostly restricted to gallery forests in the Itremo Massif, in moist substrate between rocks bordering streams. The main objective was to understand the influence of diverse mycorrhizal fungi on seed germination and further development of C. purpurea. The study aims to compare symbiotic versus asymbiotic germination and seedling development with seeds and fungi collected from a 13-km² area in the Itremo region. Seeds collected from the wild were sown with diverse orchid mycorrhizal fungi (OMF) spanning 12 operational taxonomic units (OTUs) in three genera (Tulasnella, Ceratobasidium, and Sebacina) acquired from different habitats. Treatments were assessed in terms of the percentage of germinated seeds and fully developed seedlings against those in asymbiotic control media treatments. Overall, OMF significantly improved seedling development within the 12-week experiment period. Sebacina as a genus was the most effective at promoting seedling development of C. purpurea, as well as having the ability to enter into successful symbiotic relationships with orchids of different life forms; this new knowledge may be especially useful for orchid conservation practiced in tropical areas like Madagascar. A Sebacina isolate from an epiphytic seedling of Polystachya concreta was the most effective at inducing rapid seedling development and was among the five that outperformed fungi isolated from roots of C. purpurea. C. purpurea was found to be a mycorrhizal generalist, despite its specific habitat preference, highlighting the complex interaction between the plant, fungi, and the environment. The potential impact on conservation strategies of understanding the requirements for orchid seed germination and development by identifying and using OMF from diverse sources is discussed in detail.     

Soil fungal community variation by large-scale reclamation in Sanjiang plain,China

Large-scale marshland reclamation can cause substantial changes to the soil fungal community by disturbances associated with the growth of crop plants and by the addition of fertilizers and pesticides. In this study, high-throughput sequencing of the fungal-specific internal transcribed spacer (ITS) gene region was used to identify fungal taxa. We analyzed the variation in soil fungi diversity and community composition in marshland, paddy, and farmland corn soils, and investigated the relationship between soil fungal community composition and soil physicochemical characteristics to quantify the effect of large-scale reclamation on marshland soil environment in the Sanjiang Plain, northeast China. Marshland soil contained most of the 1997 operational taxonomic units (OTUs) found across all sites (1241), while paddy soil had only 614 OTUs and farmland corn soil 817 OTUs. All reclaimed lands presented a decline in richness and diversity of soil fungi at the OTU level, and soil fungal richness was significantly different between marshland and reclaimed sites (P < 0.05), although it did not differ significantly between marshland and farmland corn sites. Additionally, soil fungal community composition showed different trends and structure after the reclamation. One-way analysis of variance showed Basidiomycota, Zygomycota, Glomeromycota, and Chytridiomycota composition differed significantly between marshland and reclaimed sites (P < 0.05). Nine dominant genera (relative abundance >1.5% in at least one site) and many unclassified genera showed significant variation between marshland and reclaimed sites, including Blumeria, Tomentella, Peziza, Hypholoma, Zopfiella, Mrakia, and Fusarium. Soil fungal community composition and diversity were affected by soil moisture, pH, total carbon (C), available nitrogen (N), soil organic carbon, soil dissolved organic carbon, and C/N (the ratio of total carbon to total nitrogen). The present results contribute to understanding the fungal community in marshland ecosystems, and the role of environmental variability as a predictor of fungal community composition.     

Composition and diversity of the bacterial community in snow leopard (<Emphasis Type="Italic">Uncia uncia</Emphasis>) distal gut

Intestinal microflora influences many essential metabolic functions, and is receiving increasing attention from the scientific community. However, information on intestinal microbiota, especially for large wild carnivores, is insufficient. In the present study, the bacterial community in the feces of snow leopards (Uncia uncia) was described based on 16S rRNA gene sequence analysis. A total of 339 near-full-length 16S rRNA gene sequences representing 46 non-redundant bacterial phylotypes (operational taxonomical units, OTUs) were identified in fecal samples from four healthy snow leopards. Four different bacterial phyla were identified: Firmicutes (56.5 %), Actinobacteria (17.5 %), Bacteroidetes (13 %), and Proteobacteria (13 %). The phylum Actinobacteria was the most abundant lineage, with 40.4 % of all identified clones, but Clostridiales, with 50 % of all OTUs, was the most diverse bacterial order. The order Clostridiales was affiliated with four families: Clostridiaceae I, Lachnospiraceae, Peptostreptococcaceae, and Ruminococcaceae. Lachnospiraceae was the most diverse family with 17 OTUs identified. These findings were basically consistent with previous reports on the bacterial diversity in feces from other mammals.     

A model for determining ex situ conservation priorities in big genera is provided by analysis of the subgenera of <Emphasis Type="Italic">Rhododendron</Emphasis> (Ericaceae)

The large size and complex taxonomy of big genera complicates decision making for conservation. We propose that Rhododendron, comprising some 1215 taxa, divided into nine subgenera and many sections, can be used as a model for other big genera. Although Red List assessments placed 715 taxa in a threat category, or listed them as Data Deficient, and moreover Target 8 of the Global Strategy for Plant Conservation requires 75 % of Red List taxa to be held in ex situ collections by 2020, to date there have been few studies of Rhododendron ex situ collections or conservation priorities. Utilising the subgenus structure of Rhododendron as a framework for examining conservation priorities, we analysed the Red List and determined that subgenera Vireya and Hymenanthes have the most acute conservation issues. Examination of taxa in cultivation shows that 844 of 1215 taxa (70 %) are in cultivation, with subgenera varying from 45 to 100 %. Of the 715 Red List taxa, 400 (56 %) are in cultivation, with subgenera varying from 28 to 72 %. Subgenera Vireya and Azaleastrum have the poorest representation in cultivation and should have precedence for ex situ conservation. As no subgenus reaches the requirement for Target 8, further planning is needed for ex situ conservation of Rhododendron. After combining the two analyses, we propose the priorities for ex situ conservation should be ordered (i) Vireya, (ii) Azaleastrum and (iii) Hymenanthes. Finally, we propose five conservation actions for Rhododendron, and summarise our approach as a model for conservation of other big genera.     

<Emphasis Type="Italic">Pyrola japonica</Emphasis>, a partially mycoheterotrophic Ericaceae,has mycorrhizal preference for russulacean fungi in central Japan

Mycorrhizal symbiosis often displays low specificity, except for mycoheterotrophic plants that obtain carbon from their mycorrhizal fungi and often have higher specificity to certain fungal taxa. Partially mycoheterotrophic (or mixotrophic, MX) plant species tend to have a larger diversity of fungal partners, e.g., in the genus Pyrola (Monotropoideae, Ericaceae). Preliminary evidence however showed that the Japanese Pyrola japonica has preference for russulacean fungi based on direct sequencing of the fungal internal transcribed spacer (ITS) region from a single site. The present study challenges this conclusion using (1) sampling of P. japonica in different Japanese regions and forest types and (2) fungal identification by ITS cloning. Plants were sampled from eight sites in three regions, in one of which the fungal community on tree ectomycorrhizal (ECM) tips surrounding P. japonica was also analyzed. In all, 1512 clone sequences were obtained successfully from 35 P. japonica plants and 137 sequences from ECM communities. These sequences were collectively divided into 74 molecular operational taxonomic units (MOTUs) (51 and 33 MOTUs, respectively). MOTUs from P. japonica involved 36 ECM taxa (96 % of all clones), and 17 of these were Russula spp. (76.2 % of all clones), which colonized 33 of the 35 sampled plants. The MOTU composition significantly differed between P. japonica and ECM tips, although shared species represented 26.3 % of the ECM tips community in abundance. This suggests that P. japonica has a preference for russulacean fungi.     

Diversity of fungi associated with roots of <Emphasis Type="Italic">Calanthe</Emphasis> orchid species in Korea

While symbiotic fungi play a key role in the growth of endangered Calanthe orchid species, the relationship between fungal diversity and Calanthe species remains unclear. Here, we surveyed root associated fungal diversity of six Calanthe orchid species by sequencing the internal transcribed spacer (ITS) region using 454 pyrosequencing. Our results revealed that Paraboeremia and Coprinopsis are dominant fungal genera among Calanthe species. In terms of overall relative abundance, Paraboeremia was the most common fungal genus associated with Calanthe roots, followed by Coprinopsis. Overall fungal diversity showed a significant degree of variation depending on both location and Calanthe species. In terms of number of different fungal genera detected within Calanthe species, C. discolor had the most diverse fungal community, with 10 fungal genera detected. This study will contribute toward a better understanding of those fungi that are required for successful cultivation and conservation of Korean Calanthe species.     

First Record of Larval Secretions of <Emphasis Type="Italic">Cochliomyia macellaria</Emphasis> (Fabricius, 1775) (Diptera: Calliphoridae) Inhibiting the Growth of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>

Maggot debridement therapy (MDT) consists on the intentional and controlled application of sterilized larvae of the order Diptera on necrotic skin lesions with the purpose of cleaning necrotic tissue and removing pathogenic bacteria. During MDT, a marked antimicrobial activity has been reported in literature specially associated with antibacterial substances from Lucilia sericata (Meigen); however, regarding Cochliomyia macellaria (Fabricius), little is known. This study aimed to evaluate in vitro inhibition of bacterial growth of Pseudomonas aeruginosa and Staphylococcus aureus in contact with excretions and secretions (ES) from C. macellaria larvae. Larval ES were extracted in sterile distilled water and divided in three groups: ES, containing 400 μL of autoclaved ES; ES+BAC, containing 400 μL of autoclaved ES+0.5-μL bacterial inoculum; and CONT-BAC, containing 400 μL of sterile distilled water +0.5 μL of bacterial inoculum. Aliquots of each experimental group were plated by spreading onto Petri dishes. Seedings were made at 0, 1, 2, 4, and 12 h after the extraction of ES. In ES+BAC groups, inhibition of S. aureus was verified between times 1 and 2 h and P. aeruginosa was inhibited between 0 and 4 h. There was no growth observed in any ES group. In the CONT-BAC groups, the number of colonies from time 4 h became countless for S. aureus and decreased for P. aeruginosa. As reported in the literature, we note here that ES have excellent bactericidal activity for both gram-positive and gram-negative bacteria, and this study shows for the first time the action of the bactericidal activity of exosecretions of C. macellaria against S. aureus and P. aeruginosa.     

Different levels of hyphal self-incompatibility modulate interconnectedness of mycorrhizal networks in three arbuscular mycorrhizal fungi within the Glomeraceae

Arbuscular mycorrhizal fungi (AMF) live in symbiosis with most plant species and produce underground extraradical hyphal networks functional in the uptake and translocation of mineral nutrients from the soil to host plants. This work investigated whether fungal genotype can affect patterns of interconnections and structural traits of extraradical mycelium (ERM), by comparing three Glomeraceae species growing in symbiosis with five plant hosts. An isolate of Funneliformis coronatus consistently showed low ability to form interconnected ERM and self-incompatibility that represented up to 21 % of hyphal contacts. The frequency of post-fusion self-incompatible interactions, never detected before in AMF extraradical networks, was 8.9 %. In F. coronatus ERM, the percentage of hyphal contacts leading to perfect hyphal fusions was 1.2–7.7, while it ranged from 25.8–48 to 35.6–53.6 in Rhizophagus intraradices and Funneliformis mosseae, respectively. Low interconnectedness of F. coronatus ERM resulted also from a very high number of non-interacting contacts (83.2 %). Such findings show that AMF genotypes in Glomeraceae can differ significantly in anastomosis behaviour and that ERM interconnectedness is modulated by the fungal symbiont, as F. coronatus consistently formed poorly interconnected networks when growing in symbiosis with five different host plants and in the asymbiotic stage. Structural traits, such as extent, density and hyphal self-compatibility/incompatibility, may represent key factors for the differential performance of AMF, by affecting fungal absorbing surface and foraging ability and thus nutrient flow from soil to host roots.     

High specificity of a rare terrestrial orchid toward a rare fungus within the North American tallgrass prairie

The Orchidaceae are globally distributed and represent a diverse lineage of obligate mycotrophic plants. Given their dependence on symbiotic fungi for germination and/or plant development, fungal community structure in substrates is expected to influence the distribution and persistence of orchid species. Yet, simultaneous characterization of orchid mycorrhizal fungal (OMF) communities in roots and in soil is rarely reported. To explain the co-distributions of OMF in roots, orchid-occupied, and bulk soil, we characterized mycorrhizal fungi associated with Platanthera praeclara over multiple years across its entire natural distribution within the North American tallgrass prairie. Root derived OMF communities included 24 Ceratobasidiaceae and 7 Tulasnellaceae operational taxonomic units (OTUs) though the orchid exhibited high spatio-temporal specificity toward a single Ceratobasidiaceae OTU, which was strongly stable across population sizes and phenological stages of the sampled individuals. The preferred OMF OTUs were primarily restricted to orchid-occupied locations while infrequent or absent in bulk soil. Variation in soil OMF assemblies was explained most by soil moisture, magnesium, manganese, and clay. In this first study of coupled root and soil OMF communities across a threatened grassland ecosystem, we report a strong relationship, further nuanced by soil chemistry, between a rare fungus and a rare orchid.     

A Polymorphism Within the 3′UTR of <Emphasis Type="Italic">NLRP3</Emphasis> is Associated with Susceptibility for Ischemic Stroke in Chinese Population

Stroke was regarded as a severe disorder with high morbidity and high mortality worldwide, ischemic stroke (IS) accounts for 85 to 90 % of new increased stroke cases. Partial mechanisms were elucidated by genetic factors including genomic instability such as single nucleotide polymorphism (SNP). Previous reports demonstrated that inflammation was involved in IS, NLRP3 [nucleotide-binding domain (NOD)-like receptor protein 3], acting as a specific inflammatory gene, however, its function and influence on IS was not well clarified. In this study, a case–control study including 1102 IS patients and 1610 healthy controls was conducted to investigate the association between IS susceptibility with a SNP (rs10754558) in 3′UTR of NLRP3. Logistic regression analysis showed that the heterozygote and the homozygote GG confer a significantly increased risk of CRC after controlling for other covariates (adjusted OR = 1.52, 95 % C.I. 1.19–1.97, P = 0.002; adjusted OR = 2.22, 95 % C.I. 2.18–3.67, P < 0.001, respectively). Carriage of G allele was associated with a greatly increased risk of developing the disease (OR = 1.69, 95 % C.I. 1.31–1.83, P < 0.001). Stratification analysis found that hypertension had interaction with rs10754558 to modulate IS risk. Further in vitro assay revealed that rs10754558 can affect mRNA level of NLRP3, suggesting its possible functional significance. Our data suggested that genetic polymorphisms in NLRP3 may influence IS risk in Chinese population. Replication of our studies in other populations and further functional studies are required for complete comprehension of the roles of NLRP3 polymorphisms in IS risk.     

Variation in Mycorrhizal Associations with Tulasnelloid Fungi among Populations of Five Dactylorhiza Species

Background

Orchid species rely on mycorrhizal symbioses with fungi to complete their life cycle. Although there is mounting evidence that orchids can associate with several fungi from different clades or families, less is known about the actual geographic distribution of these fungi and how they are distributed across different orchid species within a genus.

Methodology/Principal Findings

We investigated among-population variation in mycorrhizal associations in five species of the genus Dactylorhiza (D. fuchsii, D. incarnata, D. maculata, D. majalis and D. praetermissa) using culture-independent detection and identification techniques enabling simultaneous detection of multiple fungi in a single individual. Mycorrhizal specificity, determined as the number of fungal operational taxonomic units (OTUs), and phylogenetic diversity of fungi were compared between species, whereas discriminant analysis was used to compare mycorrhizal spectra across populations and species. Based on a 95% cut-off value in internal transcribed spacer (ITS) sequence similarity, a total of ten OTUs was identified belonging to three different clades within the Tulasnellaceae. Most OTUs were found in two or more Dactylorhiza species, and some of them were common and widespread, occurring in more than 50% of all sampled populations. Each orchid species associated with at least five different OTUs, whereas most individuals also associated with two or more fungal OTUs at the same time. Phylogenetic diversity, corrected for species richness, was not significantly different between species, confirming the generality of the observed orchid mycorrhizal associations.

Conclusions/Significance

We found that the investigated species of the genus Dactylorhiza associated with a wide range of fungal OTUs from the Tulasnellaceae, some of which were widespread and common. These findings challenge the idea that orchid rarity is related to mycorrhizal specificity and fungal distribution.     

Persistence of ecto- and ectendomycorrhizal fungi associated with <Emphasis Type="Italic">Pinus montezumae</Emphasis> in experimental microcosms

Ectomycorrhizal (ECM) and ectendomycorrhizal fungal species associated with Pinus montezumae were recorded in 8 year-old trees established in microcosms and compared with those associated with 2 year-old trees, in order to determine their persistence over the long-term. Mycorrhizal root tips were morphologically and anatomically characterized and sequenced. The extension of extramatrical mycelium of ECM fungi with long exploration strategies was evaluated. In total, 11 mycorrhizal species were registered. Seven mycorrhizal species were detected on both 2 and 8 year-old pines: Atheliaceae sp., Rhizopogon aff. fallax, R. aff. occidentalis, Suillus pseudobrevipes, Tuber separans, Wilcoxina mikolae and Wilcoxina rehmii. One species, Thelephora terrestris, was exclusively associated with two year–old seedlings, while Cenococcum geophilum, Pezizaceae sp. and Pyrenomataceae sp. were exclusively found on 8 year-old trees. Atheliaceae sp. was the ECM fungal species that presented the most abundant mycelium. Finally, we report one new fungal species of Pezizaceae occurring as a symbiont of P. montezumae.     

The relationship between adiposopathy and glucose-insulin homeostasis is not affected by moderate-intensity aerobic training in healthy women with obesity

The contribution of adiposopathy to glucose-insulin homeostasis remains unclear. This longitudinal study examined the potential relationship between the adiponectin/leptin ratio (A/L, a marker of adiposopathy) and insulin resistance (IR: homeostasis model assessment (HOMA)), insulin sensitivity (IS: Matsuda), and insulin response to an oral glucose tolerance test before and after a 16-week walking program, in 29 physically inactive pre- and postmenopausal women with obesity (BMI, 29–35 kg/m²; age, 47–54 years). Anthropometry, body composition, VO₂max, and fasting lipid-lipoprotein and inflammatory profiles were assessed. A/L was unchanged after training (p =?0.15), despite decreased leptin levels (p <?0.05). While the Matsuda index tended to increase (p =?0.07), HOMA decreased (p <?0.05) and fasting insulin was reduced (p <?0.01) but insulin area under the curve (AUC) remained unchanged (p =?0.18) after training. Body fatness and VO₂max were improved (p <?0.05) while triacylglycerols increased and HDL-CHOL levels decreased after training (p <?0.05). At baseline, A/L was positively associated with VO₂max, HDL-CHOL levels, and Matsuda (0.37?< ρ <?0.56; p <?0.05) but negatively with body fatness, HOMA, insulin AUC, IL-6, and hs-CRP levels (??0.41?< ρ <???0.66; p <?0.05). After training, associations with fitness, HOMA, and inflammation were lost. Multiple regression analysis revealed A/L as an independent predictor of IR and IS, before training (partial R² =?0.10 and 0.22), although A/L did not predict the insulin AUC pre- or post-intervention. A significant correlation was found between training-induced changes to A/L and IS (r =?0.38; p <?0.05) but not with IR or insulin AUC. Although changes in the A/L ratio could not explain improvements to glucose-insulin homeostasis indices following training, a relationship with insulin sensitivity was revealed in healthy women with obesity.     

First aeromycological study in an avocado agroecosystem in Mexico

Avocados (Persea americana Mill.) are economically and nutritionally valuable despite their susceptibility to several fungal diseases. This study was conducted in an agroecosystem of avocado trees in México. The air samples were collected every week by gravimetric methods at a height of 2 m from the ground level. Fungal colonies were isolated and transferred onto PDA and identified using morphological methods. During air sampling, different meteorological variables were measured. The symptoms of avocado diseases were determined by visual observations. To the best of our knowledge, this is the first study on aeromycological characterization in an avocado agroecosystem. Thirty-two airborne fungal genera were identified; Fusarium (97.2 %) and Colletotrichum (94.4 %) were the most common fungal pathogens present in the atmosphere of avocado. In addition, seven genera of important phytopathogenic fungi of other crops (Alternaria, Capnodium, Pestalotia, Stemphylium, Rhizopus, Curvularia, and Phyllachora) were isolated. The maximum concentration of total fungi was observed in June (358 CFU m^?3) and the minimum in September (83 CFU m^?3). The total fungal concentrations were significantly negatively correlated with the temperature. The symptoms of five diseases of avocado, viz. anthracnose, scab, spot, canker trunk, and vascular wilt, were identified in the area study. It was observed that the symptoms of all the above-mentioned diseases were present in June; and in the same month, the fungal concentrations were highest.     

First volumetric record of fungal spores in the atmosphere of Montevideo City,Uruguay: a 2-year survey

In Uruguay, aeromycological studies are restricted to a gravimetric analysis performed from December 1942 to March 1944 in Montevideo where spores of Pucciniaceae, Alternaria and Helminthosporium were the only specimens identified. Daily monitoring of airborne fungal spores was carried out for the first time in Montevideo, from April 2012 to March 2014, using a Rotorod sampler in order to evaluate the seasonal variation and influence of meteorological parameters. A total of 548,309.68 spores/m³ were recorded which belong to anamorphs of Higher Fungi (69.18 %), Phyla Ascomycota (12.62 %), Basidiomycota (8.01 %), Oomycota (0.37 %) and Myxomycota (0.06 %). Airborne spores occurred in Montevideo throughout the whole year. However, a seasonal pattern was revealed, with the highest concentrations recorded in autumn and summer. The most abundant spore types were Cladosporium (53.22 %), Alternaria (6.62 %), Didymella Group (5.86 %), Leptosphaeria Group (4.37 %) and Coprinus (4.3 %). Temperature appeared to be the most influential meteorological factor correlating significantly and positively with total spore, Cladosporium, Alternaria and Didymella Group abundance. Relative humidity influenced positively total spore, Cladosporium and Didymella Group concentrations while a weak negative association was obtained for Alternaria. Wind speed correlated negatively with total spore, Cladosporium, Alternaria and Didymella Group. Precipitation showed a negative influence on Alternaria, while positive correlations were observed for Didymella Group. For the first time, fungal spores considered allergenic were recorded in Montevideo atmosphere and the risk of exposure would have been high from December to June. However, long-term sampling is needed to define seasonal prevalence patterns and the influence of meteorological conditions on spore abundance.     

Analysis of the effect of plant growth regulators and organic elicitors on antibacterial activity of <Emphasis Type="Italic">Eucomis autumnalis</Emphasis> and <Emphasis Type="Italic">Drimia robusta</Emphasis> ex vitro-grown biomass

The effects of plant growth regulators (PGRs) and organic elicitors (OEs) on in vitro propagation of Eucomis autumnalis was established. Three-year-old ex vitro grown plants from organogenesis of E. autumnalis and somatic embryogenesis (previously reported protocol) of Drimia robusta were investigated for antibacterial activity. In vitro propagation from leaf explants of E. autumnalis was established using different PGRs and OE treatments for mass propagation, biomass production and bioactivity analysis to supplement the use of wild plant material. Prolific shoots (16.0?±?0.94 shoots per explant) were obtained with MS (Murashige and Skoog in Physiol Plant 15:473–497, 1962) medium containing 100 mg l^?1 haemoglobin (HB), 10 µM benzyladenine (BA) and 2 µM naphthaleneacetic acid (NAA). The shoots were rooted effectively with a combination of 2.5 µM indole-3-acetic acid and 5.0 µM indole-3-butyric acid. The plantlets were successfully acclimatized in a vermiculite-soil mixture (1:1 v/v) in the greenhouse. Three-year-old ex vitro-grown E. autumnalis and D. robusta plants derived via organogenesis and somatic embryogenesis respectively exhibited antibacterial activity and varied with PGR and OE treatments, plant parts and bacteria. The leaves of E. autumnalis ex vitro-derived from a combination of HB, BA and NAA followed by the individual treatments of BA and HB gave the best antibacterial activities (<?1 mg ml^?1: minimum inhibitory concentration from 0.098 to 0.78 mg ml^?1) against all tested pathogenic bacteria (Bacillus subtilis, Enterococcus faecalis, Micrococcus luteus, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa). The bulbs of D. robusta ex vitro-derived from solid culture with 10 µM picloram, 1 µM thidiazuron and 20 µM glutamine exhibited good antibacterial activity against E. faecalis, M. luteus and S. aureus when compared with other treatments and mother plants. The ex vitro-grown E. autumnalis and D. robusta biomass produced with PGRs along with OE treatments confirmed a good potent bioresource and can be used as antibacterial agents. The in vitro plant regeneration of E. autumnalis and D. robusta protocols and ex vitro plants could be used for conservation strategies, bioactivity and traditional medicinal use.