16S-23S rDNA Research Articles

Phenotypic and genotypic approach to characterize a Trueperella pecoris strain isolated from necrotic vestibulitis of a camel (Camelus dromedarius).

The present study was designed to characterize phenotypically and genotypically a Trueperella (T.) pecoris strain isolated from necrotic vestibulitis of a 10-year-old camel (Camelus dromedarius). The species identity of T. pecoris 203/7 investigated in the present study could be confirmed by phenotypic properties and by phylogenetic analyses based on partial sequencing of the 16S ribosomal RNA (rRNA) gene, the 16S-23S rDNA intergenic spacer region, the glyceraldehyde 3-phosphate dehydrogenase encoding gene gap, elongation factor Tu encoding gene tuf and the target gene rpoB encoding the β-subunit of bacterial RNA polymerase. T. pecoris strain 203/7 was grouped within the genus Trueperella in the family Arcanobacteriaceae. The 16S rRNA gene analysis showed a sequence identity of 99·9% to reference strain T. pecoris DSM 111392T . The present isolate was clearly identified as T. pecoris, the most recently described species of the genus Trueperella. Strain T. pecoris 203/7 was isolated in moderate numbers from necrotic vestibulitis of the camel and could be of some importance for the infectious process. However, the investigated strain represents the first isolation of T. pecoris from a camel.

Morphological and Molecular Characterization, and Demonstration of a Definitive Host, for Sarcocystis masoni from an Alpaca (Vicugna pacos) in China.

Simple SummarySarcocystis spp. are cyst-forming intracellular protozoan parasites characterized by a two-host prey–predator life cycle. The alpaca (Vicugna pacos) is one of the South American camelids (SACs), and in recent years, this animal was introduced to China to be raised for its meat, skin, and wool and to be kept as tourist attractions and as pets. There is considerable confusion regarding the classification and nomenclature of the species of Sarcocystis in SACs. Two Sarcocystis species, named S. auchenia and S. masoni, are currently regarded as valid in SACs based on sarcocyst morphology and 18S rDNA sequences. However, the definitive host of S. masoni remains unknown. Here, S. masoni sarcocysts in an alpaca were morphologically described and molecularly characterized. Furthermore, the life cycle of S. masoni was completed via experimental animal infection. The present analysis showed that S. masoni has a close relationship with S. cameli in the dromedary camel (Camelus dromedaries), and the relationship between the two parasites needs to be clarified in the future.Only 18S rDNA sequences of Sarcocystis spp. in South American camelids (SACs) are deposited in GenBank as references, and the definitive host of S. masoni in SACs is still unclear. Here, S. masoni sarcocysts detected in an alpaca (Vicugna pacos) in China were investigated with the aid of light (LM) and transmission electron (TEM) microscopy, and characterized using four genetic markers, i.e., 18S rDNA, 28S rDNA and ITS, and the mitochondrial cox1. Additionally, the life cycle of the parasite was completed via experimental animal infection. Under LM, S. masoni sarcocysts exhibited numerous 1.3–2.1 μm conical protrusions. Under TEM, the sarcocyst wall contained conical, cylindrical, or irregular-shaped villar protrusions, similar to type 9j. Two dogs (Canis familiaris) fed S. masoni sarcocysts shed sporocysts with a prepatent period of 8–9 days. The newly obtained 18S rDNA sequences showed 98.4–100% identity with those of S. masoni in SACs previously deposited in GenBank. Interestingly, the newly obtained sequences of 18S rDNA and mitochondrial cox1 shared 99.6–100% and 98.2–98.5% identity, respectively, with those of S. cameli in dromedary camels (Camelus dromedaries). Phylogenetic analysis based on sequences of 18S rDNA, 28S rDNA, or mitochondrial cox1 revealed that S. masoni has a close relationship with Sarcocystis spp. in ruminants. The relationship between S. masoni and S. cameli deserves to be further clarified in the future.

Production of prickly pear (Opuntia ficus-indica) vinegar in submerged culture using Acetobacter malorum and Gluconobacter oxydans: Study of volatile and polyphenolic composition

Prickly pear (Opuntia ficus-indica (L). Mill.) vinegar has been produced by submerged culture using scale laboratory fermenters at three acetification temperatures (30 °C, 37 °C, and 40 °C). Pure thermotolerant Acetic Acid Bacteria (AAB) which had been previously submitted to molecular identification by PCR–RFLP of the 16S–23S rDNA regions and 16S–23S ITS rDNA sequencing, were used. The AAB used were identified as Acetobacter malorum and Gluconobacter oxydans. The acetic fermentation achieved by A. malorum at 30 °C and 37 °C was more efficient, in terms of acidity, than that accomplished by G. oxydans. The chemical analysis revealed the presence of 85 individual volatile compounds and 17 polyphenolic compounds. The concentration of approximately half the volatile compounds was significantly affected by fermentation temperature, with clearly lower concentrations as temperature increased, whereas few significant differences were observed when comparing the vinegars produced by the two AAB species. Regarding phenolic compounds, significant increases were registered when temperature changed from 30 °C to 40 °C. Furthermore, the vinegars produced by A. malorum presented a greater concentration of phenolic compounds than those produced by G. oxydans.

Reappraisal of theCrangonyx floridanusspecies complex, with the description of a new species ofCrangonyxBate, 1859 (Amphipoda: Crangonyctidae) from northern Florida, USA

AbstractCrangonyx apalachee n. sp. Cannizzaro & Sawicki in Cannizzaro, Sisco & Sawicki, is described from Lake Jackson in Leon County, Florida, USA, a prairie lake that periodically drains through karst sinkholes during times of low rainfall. The new species occurs in tall grass along the lake’s shoreline. Molecular analyses of the nuclear 18S rDNA and 28S rDNA along with the mitochondrial 16S rDNA, combined with morphological analyses were used to delineate the population from closely related crangonyctids. The occurrence of C. apalacheen. sp. in Lake Jackson marks the fifth species to be described from the floridanus complex in Florida. Collections from other locations across north Florida suggest the presence of additional floridanus-complex species still to be described.

Two New Species and One New Genus of Glass Sponges (Hexactinellida: Euplectellidae and Euretidae), From a Transect on a Seamount in the Northwestern Pacific Ocean

Hexactinellid sponges often form structural habitats for other organisms and thus support high biodiversity. Two representative morphotypes of hexactinellid sponges were sampled by a remotely operated vehicle along a transect (depths of 2377–2758 m) on the Ko-Hakucho Guyot in the northwestern Pacific Ocean, both new to science. One new species,Flavovirens hemiglobusgen. et sp. nov., which is pedunculate and bears the main choanosomal spicules of diactins, is clearly attributed to the euplectellid subfamily Bolosominae. Its set of microscleres present (namely, predominating oxyhexasters and discasters, and rare discohexasters and staurodiscs) characterizes it as a new genus. The other new species,Chonelasma tyloscopulatumsp. nov., which is funnel in form without dichotomous branching tubes or lateral oscula and has firm body walls supported by a three-layered dictyonal framework, is clearly attributed to the euretid genusChonelasma(subfamily Chonelasmatinae). Its presence of surface pentactins, two types of scopules, and both oxy- and disco-tipped hexasters or hemihexasters as common microscleres, characterizes it as a new species. The placements are also supported by molecular phylogenetic evidence from nuclear 18S rDNA and 28S rDNA, and mitochondrial 16S rDNA and cytochrome c oxidase subunit I (COI) gene. More sampling efforts should be conducted to improve the understanding of the biodiversity of deep-sea seamount sponges.

Uneven Levels of 5S and 45S rDNA Site Number and Loci Variations across Wild Chrysanthemum Accessions.

Ribosomal DNA (rDNA) is an excellent cytogenetic marker owing to its tandem arrangement and high copy numbers. However, comparative studies have focused more on the number of rDNA site variations within the Chrysanthemum genus, and studies on the types of rDNA sites with the same experimental procedures at the species levels are lacking. To further explore the number and types of rDNA site variations, we combined related data to draw ideograms of the rDNA sites of Chrysanthemum accessions using oligonucleotide fluorescence in situ hybridization (Oligo-FISH). Latent variations (such as polymorphisms of 45S rDNA sites and co-localized 5S-45S rDNA) also occurred among the investigated accessions. Meanwhile, a significant correlation was observed between the number of 5S rDNA sites and chromosome number. Additionally, the clumped and concentrated geographical distribution of different ploidy Chrysanthemum accessions may significantly promote the karyotype evolution. Based on the results above, we identified the formation mechanism of rDNA variations. Furthermore, these findings may provide a reliable method to examine the sites and number of rDNA variations among Chrysanthemum and its related accessions and allow researchers to further understand the evolutionary and phylogenetic relationships of the Chrysanthemum genus.

Rhizobial diversity is associated with inoculation history at a two-continent scale.

A total of 120 Mesorhizobium strains collected from the central dry zone of Myanmar were analyzed in a pot experiment to evaluate nodulation and symbiotic effectiveness (SE%) in chickpea plants. Phylogenetic analyses revealed all strains belonged to the genus Mesorhizobium according to 16–23S rDNA IGS and the majority of chickpea nodulating rhizobia in Myanmar soils were most closely related to M. gobiense, M. muleiense, M. silamurunense, M. tamadayense and M. temperatum. Around two-thirds of the Myanmar strains (68%) were most closely related to Indian strain IC-2058 (CA-181), which is also most closely related to M. gobiense. There were no strains that were closely related to the cognate rhizobial species to nodulate chickpea: M. ciceri and M. mediterraneum. Strains with diverse 16S–23S rDNA IGS shared similar nodC and nifH gene sequences with chickpea symbionts. Detailed sequence analysis of nodC and nifH found that the strains in Myanmar were somewhat divergent from the group including M. ciceri and were more closely related to M. muleiense and IC-2058. A cross-continent analysis between strains isolated in Australia compared with Myanmar found that there was little overlap in species, where Australian soils were dominated with M. ciceri, M. temperatum and M. huakuii. The only co-occurring species found in both Myanmar and Australia were M. tamadayense and M. silumurunense. Continued inoculation with CC1192 may have reduced diversity of chickpea strains in Australian soils. Isolated strains in Australian and Myanmar had similar adaptive traits, which in some cases were also phylogenetically related. The genetic discrepancy between chickpea nodulating strains in Australia and Myanmar is not only due to inoculation history but to adaptation to soil conditions and crop management over a long period, and there has been virtually no loss of symbiotic efficiency over this time in strains isolated from soils in Myanmar.

Morphological and Molecular Description of Sarcocystis myodes n. sp. from the Bank Vole (Clethrionomys glareolus) in Lithuania

Simple SummaryRepresentatives of genus Sarcocystis (Apicomplexa, Sarcocystidae) are parasites of mammals, birds, and reptiles. They are characterized by two-host prey-predator life cycle. Rodents are reservoirs of zoonotic diseases and play a significant role in a spread of pathogens. At present, about 40 Sarcocystis species are known to form sarcocysts in muscles and brain of rodents. Most of Sarcocystis spp. in these hosts have been characterized by morphological methods and life cycle investigations. In the present study a new Sarcocystis species, S. myodes is described in skeletal muscles of the bank voles (Clethrionomys glareolus) from Lithuania using morphological, molecular, and phylogenetic analysis. Based on five genetic loci, S. myodes was most closely related to Sarcocystis spp. using predatory mammals as their definitive hosts. The analysis of previous studies indicates that Sarcocystis spp. diversity in voles is not fully revealed. Furthermore, some Sarcocystis spp. formerly detected in voles are important pathogens. Therefore, further molecular examinations are needed for the revision of Sarcocystis spp. in these hosts.Numerous rodent species have been broadly examined for Sarcocystis parasites. Nevertheless, recent investigations on Sarcocystis spp. in voles are lacking. As many as 45 bank voles (Clethrionomys glareolus) captured in several locations in Lithuania were examined in the present study. Based on morphological, genetic, and phylogenetic results, sarcocysts detected in one bank vole were described as Sarcocystis myodes n. sp. Using light microscopy analysis, the observed sarcocysts were ribbon-shaped, 6000–3000 × 70–220 µm in size. Sarcocysts were characterized by a relatively thin (about 1 μm) and apparently smooth cyst wall. The lancet-shaped bradyzoites were 9.6–12.0 × 3.1–4.6 μm in size. By transmission electron microscopy, the sarcocyst wall was up to 1 μm thick, parasitophorous vacuolar membrane had small knob-like blebs. Based on 18S rDNA, 28S rDNA, cox1, rpoB, and ITS1 loci, S. myodes showed highest similarity with S. ratti from the black rat (Rattus rattus). According to phylogenetic placement, S. myodes was most closely related to Sarcocystis spp. that employ predatory mammals as their definitive hosts. Morphologically, sarcocysts of S. myodes have similar features to those of S. cernae, S. dirumpens, and S. montanaensis described in voles, however, they use birds of prey or snakes as their definitive hosts.

Distribution patterns of rDNA loci in the Schedonorus-Lolium complex (Poaceae).

The Schedonorus-Lolium complex of the subtribe Loliinae (Poaceae) includes several economically important forage and turf grasses. This complex encompasses Lolium Linnaeus, 1753, Festuca Linnaeus, 1753 subgenus Schedonorus (P. Beauvois, 1824) Petermann, 1849 and Micropyropsis Romero Zarco et Cabezudo, 1983. New FISH results of 5S and 18S–26S rDNA sequences are presented for three species and the results are interpreted in a review of distribution patterns of 5S and 18S–26S rDNA sequences among other species in the complex. Micropyropsistuberosa Romero Zarco et Cabezudo, 1983 (2n = 2x = 14) displayed a distribution pattern of rDNA sequences identical to that of F.pratensis Hudson, 1762, supporting a close phylogenetic relationship at the bottom of the phylogenetic tree. “Loliummultiflorum” Lamarck, 1779 accessions sourced from Morocco showed a different pattern from European L.multiflorum and could be a unique and previously uncharacterised taxon. North African Festucasimensis Hochstetter ex A. Richard, 1851 had a marker pattern consistent with allotetraploidy and uniparental loss of one 18S–26S rDNA locus. This allotetraploid has previously been suggested to have originated from a hybrid with Festucaglaucescens (Festucaarundinaceavar.glaucescens Boissier, 1844). However, the distribution patterns of the two rDNA sequences in this allotetraploid do not align with F.glaucescens, suggesting that its origin from this species is unlikely. Furthermore, comparisons with other higher alloploids in the complex indicate that F.simensis was a potential donor of two sub-genomes of allohexaploid Festucagigantea (Linnaeus) Villars, 1787. In the overall complex, the proximal locations of both rDNA markers were conserved among the diploid species. Two types of synteny of the two markers could, to a considerable extent, distinguish allo- and autogamous Lolium species. The ancestral parentage of the three Festuca allotetraploids has not yet been determined, but all three appear to have been sub-genome donors to the higher allopolypoids of sub-genus Schedonorus. Terminal locations of both the markers were absent from the diploids but were very frequently observed in the polyploids.

Diatom Biodiversity and Speciation Revealed by Comparative Analysis of Mitochondrial Genomes

Diatoms (Bacillariophyta) constitute one of the most diverse and ecologically significant groups of phytoplankton, comprising 100,000–200,000 species in three classes Bacillariophyceae, Mediophyceae, and Coscinodiscophyceae. However, due to the limited resolution of common molecular markers including 18S rDNA, 28S rDNA, ITS, rbcL, and cox1, diatom biodiversity has not been adequately ascertained. Organelle genomes including mitochondrial genomes (mtDNAs) have been proposed to be “super barcodes” for distinguishing diatom species because of their rich genomic content, and the rapid progress of DNA sequencing technologies that has made it possible to construct mtDNAs with increasing throughout and decreasing cost. Here, we constructed complete mtDNAs of 15 diatom species including five Coscinodiscophyceae species (Guinardia delicatula, Guinardia striata, Stephanopyxis turris, Paralia sulcata, and Actinocyclus sp.), four Mediophyceae species (Hemiaulus sinensis, Odontella aurita var. minima, Lithodesmioides sp., and Helicotheca tamesis), and six Bacillariophyceae species (Nitzschia ovalis, Nitzschia sp., Nitzschia traheaformis, Cylindrotheca closterium, Haslea tsukamotoi, and Pleurosigma sp.) to test the practicality of using mtDNAs as super barcodes. We found that mtDNAs have much higher resolution compared to common molecular markers as expected. Comparative analysis of mtDNAs also suggested that mtDNAs are valuable in evolutionary studies by revealing extensive genome rearrangement events with gene duplications, gene losses, and gains and losses of introns. Synteny analyses of mtDNAs uncovered high conservation among species within an order, but extensive rearrangements including translocations and/or inversions between species of different orders within a class. Duplication of cox1 was discovered for the first time in diatoms in Nitzschia traheaformis and Haslea tsukamotoi. Molecular dating analysis revealed that the three diatom classes split 100 Mya and many diatom species appeared since 50 Mya. In conclusion, more diatom mtDNAs representing different orders will play great dividends to explore biodiversity and speciation of diatoms in different ecological regions.

Bacteria isolated from Triticum monococcum ssp. monococcum roots can improve wheat hologenome in agriculture.

Triticum monococcum ssp. monococcum is an ancestral wheat species originated from Karacadağ Mountain of Turkey more than ten thousand years ago. Because of environmental and anthropogenic effects, food supply and demand are not balanced. Agricultural activities such as breeding, and fertilization are important to sustain the balance. Conventional breeding and fertilization applications usually neglect contribution of plant related hologenomes in agricultural yield. The disruption of plant growth promoting microorganisms results in intensive usage of chemical fertilizers. The harmony between plant and plant-associated microorganisms is important for sustainability. In this study, isolation, biochemical characterization, and impact on plant growth parameters of natural bacteria associated with Triticum monococcum ssp. monococcum hologenome were aimed. The collection of root samples and isolations of the root-associated bacterial species were carried out from local wheat lands. According to interpretation of three identification methods (MALDI-TOF, 16S rDNA, 16S-23S rDNA) eight isolates are Arthrobacter spp. ESU164, Arthrobacter spp. ESU193, Pseudomonas spp. ESU131, Pseudomonas spp. ESU141, Pseudomonas poae strain ESU182, Pseudomonas thivervalensis strain ESU192, Pseudomonas spp. ESU1531, Bacillus subtilis strain ESU181. For each isolate we investigated biochemical properties especially nitrogen fixation, phosphate solubilization, and indole-3-acetic acid production abilities. The results show that all isolates are nitrogen fixers and the best phosphate solubilizer have been reported as Pseudomonas spp. ESU131 with 2.805 ± 0.439. All isolates are indole-3-acetic acid productors. 2 isolates affected the coleoptile lengths, 7 bacterial isolates showed statistically positive effect on root number, and 5 isolates promote the root lengths and the root fresh weights.

Phylogenetic position of Acanthobothrium cleofanus (Cestoda: Onchoproteocephalidea) using molecular evidence.

Despite the large number of species described to date for the onchoprotepcephalid genus Acanthobothrium (207), only 16 named species have a genetic sequence. With this background, specimens of adult cestodes of the stingray Hypanus longus were collected off San Blas, Nayarit, and onchoproteocephalid larvae in the carangid fish Trachinotus rhodopus from Puerto Ángel, Oaxaca, both located on the Pacific coast of Mexico. The objective of this work is to investigate the phylogenetic position of these adults and larvae using nuclear ribosomal markers (18S rDNA and 28S rDNA). Morphologically, adult specimens were identified as Acanthobothrium cleofanus; larvae were identified only to family level. The phylogenetic position of both taxa was investigated based on the information of two nuclear molecular markers analyzed under Parsimony (PA) and Bayesian Inference (BI) methods. The newly generated sequences of A. cleofanus from Nayarit are identical to the sequences of several samples of Acanthobothrium sp. collected in the Mexican Pacific, which sequence are available in GenBank; DNA sequences obtained from onchoproteocephalid larva clearly place this taxon within Acanthobothrium but representing an independent lineage. In the resulting phylogenetic trees, Uncibilocularis okei was found nested within Acanthobothrium with an unstable position depending on the optimality criteria, indicating the need for more molecular analyzes with a greater number of species of both genera prior to define its phylogenetic relationships.

MOLECULAR CHARACTERIZATION OF RHIZOBIA STRAINS ASSOCIATED WITH MUNGBEAN (VIGNA RADIATA), IN SOILS OF SUDANO-GUINEAN AND SAHELIAN AGROECOLOGICAL ZONES OF MALI

In the Sudano-Guinean and Sahelian zones of Mali, the decline in soil fertility is a key challenge for a sustainable enhancement of agricultural productivity. In addition, studies on the diversity of bacteria associated with Mungbeans are rare, with very few studies conducted on their identification and molecular characterization in the soils of the major agricultural production basins in the Sudano-Guinean and Sahelian Guinean zones of Mali. This study was therefore motivated not only by these different challenges of productivity and soil fertility, but also by the numerous nutritional and agronomic interests of mung beans and the possibility of using its associated bacteria as bio-fertilizers. It aims at contributing to the improvement of mung bean (Vigna radiata L) productivity and the promotion of its cultivation using inocula based on indigenous and efficient bacteria. To achieve this objective, soil samples were taken from various farmers' fields in the two studied areas. The Physico-chemical characteristics of the soils were determined, and trapping trials were conducted in the greenhouse on these soils with Mung bean as the trap plant. The nodules formed on the plants were harvested, the rhizobia contained in these nodules were isolated, the IGS of the 16S-23S rDNA of the isolates was analyzed using PCR-RFLP, and an inoculation test was carried out in a growth chamber to verify the infectivity of the isolates. Eighty-three (83) isolates divided into two groups according to their color (whitish, yellowish) and three (03) IGS types (I, II, III) were encountered with a large dominance of IGS type I in the Guinean zone and that of IGS types I and II in the Sahel zone. Two (02) isolates of type IGS I showed a higher infectivity. These results show a low diversity of rhizobium strains associated with Mung bean in the studied soils. It would be interesting to extend this study to other areas of Mali for strains that associate with Mung bean.

Study of the diversity of 16S-23S rDNA internal transcribed spacer (ITS) typing of Escherichia colistrains isolated from various biotopes in Tunisia.

We investigated the 16S-23S rRNA intergenic spacer region (ISR)-PCR and the phylogenetic PCR analyses of 150 Escherichia coli isolates as tools to explore their diversity, according to their sampling origins, and their relative dominance in these sampling sources. These genetic markers are used to explore phylogenetic and genetic relationships of these 150 E. coli isolates recovered from different environmental sources (water, food, animal, human and vegetables). These isolates are tested for their biochemical pattern and later genotyped through the 16S-23S rRNA intergenic spacer PCR amplification and their polymorphism investigation of PCR-amplified 16S-23S rDNA ITS. The main results of the pattern band profile revealed one to four DNA fragments. Distributing 150 E. coli isolates according to their ITS and using RS-PCR, revealed four genotypes and four subtypes. The DNA fragment size ranged from 450 to 550bp. DNA band patterns analysis revealed considerable genetic diversity in interspecies. Thus, the 450 and 550bp sizes of the common bands in all E. coli isolates are highly diversified. Genotype I appeared as the most frequent with 77.3% (116 isolates), genotype II with 12% (18 isolates); genotype III with 9.7% (14 isolates), and the IV rarely occurred with 4% (2 isolates). Distributing the E. coli phylogroups showed 84 isolates (56%) of group A, 35 isolates (23.3%) of group B1, 28 isolates (18.7%) of group B2 and only three isolates (2%) of group D.

Gammarinema scyllae sp. n. and Monhystrium mangrovi sp. n. (Nematoda: Monhysteridae) from land crabs from New Caledonia

Two new species of nematodes associated with crabs are described from New Caledonia. Gammarinema scyllaesp. n. from the gill chambers of Scylla serrata (Forsskål) is characterised by 3–4 mm long body, small outer labial and cephalic sensilla, distinct ocelli, short straight spicules and sub-cylindrical tail. Monhystrium mangrovisp. n. from the gill chambers and body cavity of mangrove crab Neosarmatium sp. is characterised by 1–1.4 mm long body; outer labial sensilla longer than cephalic sensilla, amphid located at level with posterior stoma chamber, denticles in posterior stoma chamber and five pairs of genital papilla on tail. Phylogenetic relationships of two new species and other nematodes from the family Monhysteridae are analysed based on 18S and partial 28S rDNA sequences.

Comparative chromosome studies in species of subtribe Orchidinae (Orchidaceae).

In our study, FISH mapping using 18S-5.8S-25S rDNA and 5S rDNA sequences was performed for the first time on Ophrystenthredinifera Willdenow, 1805, Serapiasvomeracea (Burman f., 1770) Briquet, 1910 and Himantoglossumhircinum (Linnaeus, 1753) Sprengel, 1826. A detailed study was also performed on O.tenthredinifera using Giemsa-staining, silver-staining, CMA fluorescence banding and fluorescence in situ hybridisation (FISH) with rDNA probes. We analysed two subspecies, i.e. O.tenthrediniferasubsp.neglecta (Parlatore, 1860) E.G. Camus, 1908 and O.tenthrediniferasubsp.grandiflora (Tenore, 1819) Kreutz, 2004 by the traditional Feulgen method and constructed the karyotype. The cytotaxonomic implications for both taxa are also discussed. In Himantoglossumhircinum, FISH and silver staining highlighted differences in the number of two rDNA families (35S and 5S) with respect to Barliarobertiana (Loiseleur-Deslongchamps, 1807) Greuter, 1967. In addition, fluorescence in situ hybridisation was also applied to diploid (2n = 2x = 36) and triploid (2n = 3x = 54) Anacamptismorio (Linnaeus, 1753) R.M. Bateman, Pridgeon et M.W. Chase, 1997. As far as we are aware, this is the first case of autotriploidy observed in A.morio.

A new genetic linkage map in einkorn wheat (Triticum monococcum) detects two major QTLs for heading date in chromosome 2A and 5A, probably corresponding to the photoperiod and vernalization genes

AbstractEinkorn harbours a wide range of genetic diversity for agronomically important traits. Heading date is one of the important traits for the adaptation of wheat species to various climatic conditions and the cultivation in diverse regions and seasons. In this study, DArTseq and SSR marker‐based linkage map of einkorn wheat was constructed using 150 recombinant inbred lines (RILs) derived from a cross between ID1623 and MONLIS. The linkage map involved 865 SNP, 787 Silico‐DArT and 16 SSR DNA markers mapped to 741 loci in seven linkage groups and covered 1158.54 cM with an average density of 1.56 cM/locus. Four QTLs associated with heading date, detected in all environments by composite interval mapping (CIM), were positioned across the 2A, 5A and 7A einkorn wheat chromosomes. Two major and stable QTLs, QThd‐2A.2 and QThd 5A.1, were identified on chromosomes 2A and 5A, respectively. These QTLs probably coincide with photoperiod and vernalization genes in accordance with previously published maps. The new map will facilitate quantitative trait locus mapping, map‐based cloning and comparative genomics in wheat species.

Prevalence, Enterotoxigenic Potential and Antimicrobial Resistance of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus (MRSA) Isolated from Algerian Ready to Eat Foods.

Staphylococcus aureus causes a foodborne intoxication due to the production of enterotoxins and shows antimicrobial resistance, as in the case of methicillin-resistant strains (MRSA). Herein, we analyzed 207 ready-to-eat foods collected in Algeria, reporting a S. aureus prevalence of 23.2% (48/207) and respective loads of coagulase positive staphylococci (CPS) ranging from 1.00 ± 0.5 to 5.11 ± 0.24 Log CFU/g. The 48 S. aureus isolates were widely characterized by staphylococcal enterotoxin gene (SEg)-typing and 16S-23S rDNA intergenic spacer region (ISR)-PCR, as well as by detecting tst and mecA genes, genetic determinants of toxic shock syndrome toxin-1 and methicillin resistance, respectively. We found that the S. aureus isolates belonged to seven different SEg-types harboring the following combinations of genes: (1) selW, selX; (2) egc (seG, seI, seM, seN, seO), selW, selX; (3) seA, seH, seK, seQ, selW, selX; (4) seB, selW, selX; (5) seD, selJ, seR, selW, selX; (6) seH, selW, selX, selY; and (7) seA, egc, selW, selX, while among these, 2.1% and 4.2% were tst- and mecA- (staphylococcal chromosomal cassette mec-type IV) positive, respectively. Selected strains belonging to the 12 detected ISR-types were resistant towards antimicrobials including benzylpenicillin, ofloxacin, erythromycin, lincomycin, tetracyclin, kanamycin, oxacillin, and cefoxitin; 8.3% (1/12) were confirmed as MRSA and 16.7% (2/12) were multidrug resistant. The present study shows the heterogeneity of the S. aureus population in Algerian ready-to-eat foods as for their toxigenic potential and antimicrobial resistance, shedding the light on the quality and safety related to the consume of ready-to-eat foods in Algeria.

Pseudopolydora (Annelida: Spionidae) from the Arabian Gulf, Kuwait

Seven species of Pseudopolydora are described and illustrated from the Arabian Gulf, Kuwait: P. achaeta Radashevsky & Hsieh, 2000, P. antennata (Claparède, 1868), P. arabica Radashevsky & Al-Kandari, 2020, P. auha sp. nov., P. kuwaiti sp. nov., P. melanopalpa sp. nov., and P. multispinosa sp. nov. The morphology of the developed planktonic larvae is described for P. antennata and P. kuwaiti sp. nov. Adults of all species live in tubes in soft sediments, while adults of P. kuwaiti sp. nov. also bore in shells of gastropods and dead corals encrusted by coralline algae. Pseudopolydora antennata and P. arabica form dense settlements up to 50 000 individuals per 1 m2, while other species are comparatively rare. The phylogenetic relationships between the examined species and other Pseudopolydora (18 species in total) were assessed in an analysis of sequence data of four gene fragments: mitochondrial 16S rDNA, nuclear 18S rDNA and 28S rDNA, and Histone 3 (2473 bp in total). Three species complexes are distinguished involving P. antennata, P. diopatra Hsieh, 1992 and P. paucibranchiata (Okuda, 1937). Sets of adult morphological features shared by species of each complex and an identification key to Pseudopolydora species from the Arabian Gulf are provided.

A potential probiotic Leuconostoc mesenteroides TBE-8 for honey bee

An isolated bacterium TBE-8, was identified as Leuconostoc mesenteroides according to the sequences of 16S rDNA and the 16S–23S rDNA intergenic spacer region. The probiotic properties of the L. mesenteroides TBE-8 strain were characterized and revealed that TBE-8 could utilize various carbohydrates, exhibited high tolerance to sucrose’s osmotic pressure and acidic conditions, and could mitigate the impact of the bee pathogen Paenibacillus larvae. In addition, we found that the TBE-8 broth increased the expression of the nutrition-related genes major royal jelly protein 1 and vitellogenin in bees by approximately 1400- and 20-fold, respectively. The expression of genes encoding two antibacterial peptides, hymenoptaecin and apidaecin, in the bee abdomen was significantly increased by 17- and 7-fold in bees fed with the TBE-8 fermented broth. Furthermore, we fed four-frame bee colonies with 50% sucrose syrup containing TBE-8 and can detect the presence of approximately 2 × 106 16S rDNA copies of TBE-8 in the guts of all bees in 24 h, and the retention of TBE-8 in the bee gut for at least 5 days. These findings indicate that the L. mesenteroides TBE-8 has high potential as a bee probiotic and could enhance the health of bee colonies.