DNA sequence of the araBAD-araC controlling region in Salmonella typhimurium LT2

The araB and araC genes of Salmonella typhimurium have been cloned onto the plasmid pBR322. Restriction analysis and subcloning of restriction fragments localized these genes to a 4.4 kb DNA fragment. Complementation analysis revealed that the cloned araB and araC genes from S. typhimurium complemented araB and araC mutant strains of Escherichia coli. Conversely, cloned araB and araC genes from E. coli complemented araB and araC mutant strains of S. typhimurium. The DNA sequences was determined for the S. typhimurium araB and araC controlling region and for the initially translated portions of these genes. The nucleotide sequence of the araB promoter was 87% homologous with the same region in E. coli and contained no deletions or insertions relative to the E. coli sequence. The presumed AUG codon corresponding to the amino terminus of the S. typhimurium araC protein was in the same location as in E. coli. There was, however, considerable divergence from the E. coli sequence preceding the translation start site. The nucleotide sequence of the initial 237 bp in the open reading frame of the S. typhimurium araC gene was 78% homologous with the same sequence in E. coli. By comparison, the amino acid sequence for this region was 91% conserved.     

Molecular phylogeny of Phoma and allied anamorph genera: Towards a reclassification of the Phoma complex

The present generic concept of Phoma is broadly defined, with nine sections being recognised based on morphological characters. Teleomorph states of Phoma have been described in the genera Didymella, Leptosphaeria, Pleospora and Mycosphaerella, indicating that Phoma anamorphs represent a polyphyletic group. In an attempt to delineate generic boundaries, representative strains of the various Phoma sections and allied coelomycetous genera were included for study. Sequence data of the 18S nrDNA (SSU) and the 28S nrDNA (LSU) regions of 18 Phoma strains included were compared with those of representative strains of 39 allied anamorph genera, including Ascochyta, Coniothyrium, Deuterophoma, Microsphaeropsis, Pleurophoma, Pyrenochaeta, and 11 teleomorph genera. The type species of the Phoma sections Phoma, Phyllostictoides, Sclerophomella, Macrospora and Peyronellaea grouped in a subclade in the Pleosporales with the type species of Ascochyta and Microsphaeropsis. The new family Didymellaceae is proposed to accommodate these Phoma sections and related anamorph genera. The present study demonstrated that Phoma radicina, the type species of Phoma sect. Paraphoma and Phoma heteromorphospora, the type species of Phoma sect. Heterospora can be assigned to the Phaeosphaeriaceae and Leptosphaeriaceae respectively.     

Molecular phylogeny of Plumbaginaceae with emphasis on Acantholimon Boiss. based on nuclear and plastid DNA sequences in Iran

The present study represents phylogenetic analyses of Plumbaginaceae with emphasis on Acantholimon from Iran using nrDNA ITS and plastid trnY-T sequences. The analyses support the monophyly and the close relationship of Limonium, Armeria and Psylliostachys. This is the first report of the close relationship between Acantholimon and Cephalorhizum. The data for the position of Cephalorhizum is unclear. The Shimodaira–Hasegawa test of nrDNA ITS and the combined datasets indicated that Acantholimon and Cephalorhizum are distinct genera. The molecular data revealed that the traditionally recognized multi-specific Acantholimon sections (Acantholimon, Acmostegia, Glumaria, Microstegia, Staticopsis and Tragacanthina) are not monophyletic. Their members are intermixed with each other and scattered across the Acantholimon clade, but the smaller sections including Platystegia and Pterostegia, each comprising two species, are monophyletic.     

Re-interpreting the biochronology of the La Celia and Los Gargantones mammal sites (Late Miocene,Murcia, Spain)

The biochronological age of the small-mammal populations of Los Gargantones 1, 2 and La Celia (upper Miocene, La Celia sub-basin, Murcia, Spain) is re-interpreted. The presence in Los Gargantones of Occitanomys adroveri, Parapodemus barbarae, Parapodemus cf. gaudryi, Huerzelerimys turoliensis, Atlantoxerus cf. adroveri, and Alilepus evidences a correlation to MN12 (∼7.5–7 Ma) rather than to MN11 (∼9–7.5 Ma), as inferred previously. The assemblage corresponds to that of the more eastern, near-coast sites of Crevillente 8 and 15, situated in the Alicante area. The stratigraphically highest site of La Celia contains Hispanomys adroveri, a species also indicative of MN12. The presence of Castromys cf. littoralis together with an advanced O. adroveri points to a slightly younger age than that of Los Gargantones, approaching that of MN13 sites. The assemblage best matches that of Crevillente 17. Other species described in this paper are Prolagus crusafonti, Prolagus sp., Parasorex cf. ibericus, Panelimnoecus cf. repenningi, and Blarinella aut Petenyia sp. indet.     

The rearranged mitochondrial genome of Podagrion sp. (Hymenoptera: Torymidae), a parasitoid wasp of mantis

The complete sequence of the mitochondrial genome of Podagrion sp. (Hymenoptera: Torymidae) is described. The mitogenome was 15,845 bp in size, and contained typical sets of mitochondrial genes. The base composition of the Podagrion sp. mitogenome was also biased toward A + T bases (81.8%). The mitochondrial genome of Podagrion sp. has a weak AT skew (0.07) and a strong GC skew (?0.26). Podagrion sp. exhibits a novel rearrangement compared with the ancestral order, including six protein-coding genes (nad3, cox3, atp6, atp8, cox2 and cox1), which have inverted to the minor strand from the major strand. The A + T-rich region of Podagrion sp., which is located between trnN and trnI, have five tandem repeats. The apomorphic rearrangements, including the conserved block “cox3-atp6-atp8-cox2-cox1-nad5-nad4-nad4l-nad6-cob” and the special locations of trnV and trnA, were mapped onto the phylogeny of Proctotrupomorpha.     

Assessment of biological colonization of historic buildings in the former Auschwitz II-Birkenau concentration camp

The objective of this study was to assess biological colonization of wooden and brick buildings in the former Auschwitz II-Birkenau concentration camp, and to identify the organisms colonizing the examined buildings. Microbiological analysis did not reveal increased microbial activity, and the total microbial count of the barrack surfaces did not exceed 10³ CFU/100 cm². However, certain symptoms of biodegradation of the buildings were observed. The predominant microflora consisted of bacteria of the genera Bacillus, Sporosarcina, Pseudomonas, Micrococcus, Streptomyces, and Staphylococcus, as well as fungi of the genera Acremonium, Cladosporium, Alternaria, Humicola, Penicillium, and Chaetomium. The microflora patterns varied both in wooden and brick buildings. The structural elements of wooden and brick barracks, and especially of the floors and lower parts of bathroom walls, were infected by cyanobacteria and algae, with the most numerous being cyanobacteria of the genera Scytonema, Chroococcus, Gloeothece, Leptolyngbya, diatoms of the genus Diadesmis, and chlorophytes of the genera Chlorella and Apatococcus. The outer surfaces of the examined buildings were primarily colonized by lichens and bryophytes, with nearly 30 species identified. The dominant species of lichens belonged to the genera Candelariella, Caloplaca, Lecanora, Lecidea, Lepraria, Physcia, and Protoparmeliopsis, and those of bryophytes to the genera Bryum, Ceratodon, Marchantia, and Tortula. The quantity and species diversity of lichens and mosses were much lower in wooden barracks than in brick ones. The external surfaces of those barracks were only affected by Lecanora conizaeoides, Lecanora symmicta, Lepraria cf. incana, and Strangospora pinicola. The study results revealed vast biodiversity among the species colonizing historic buildings. The presence of these groups of organisms, resulting from their natural expansion in the environment, is undesirable, as their excessive growth and spread may lead to progressive biodegradation of buildings. Our assessment of biological contamination will enable the development of a disinfection and conservation plan for the examined buildings.     

Chemical and enzymatic synthesis of monomeric procyanidins (leucocyanidins or 3′,4′,5,7-tetrahydroxyflavan-3,4-diols) from (2R,3R)-dihydroquercetin

The major product from the reduction of (2R,3R)-dihydroquercetin with sodium borohydride is the 2,3-trans-3,4-trans isomer of leucocyanidin [(2R,3S,4R-3,3′,4,4′,5,7-hexahydroxyflavan] whereas the enzymatic reduction product is the 2,3-trans-3,4-cis isomer [(2R,3S,4S)-3,3′,4,4′,5,7-hexahydroxyflavan]. The 3,4-trans isomer may be partly converted to the 3,4-cis isomer under mild acid conditions. The 3,4-cis isomer is more acid-labile, and more reactive both chemically with thiols and enzymatically with a diol reductase, than the 3,4-trans isomer.     

Systematics of the Neotropical genus Catharylla Zeller (Lepidoptera,Pyralidae s. l., Crambinae)

The Neotropical genus Catharylla Zeller, 1863 (type species: Crambus tenellus Zeller, 1839) is redescribed. Catharylla contiguella Zeller, 1872, C. interrupta Zeller, 1866 and Myelois sericina Zeller, 1881, included by Munroe (1995) in Catharylla, are moved to Argyria Hübner. Catharylla paulella Schaus, 1922 and C. tenellus (Zeller, 1839) are redescribed. Six new species are described by Léger and Landry: C. bijuga, C. chelicerata, C. coronata, C. gigantea, C. mayrabonillae and C. serrabonita. The phylogenetic relationships were investigated using morphological as well as molecular data (COI, wingless, EF-1α genes). The median and subterminal transverse lines of the forewing as well as the short anterior and posterior apophyses of the female genitalia are characteristic of the genus. The monophyly of Catharylla was recovered in all phylogenetic analyses of the molecular and the combined datasets, with three morphological apomorphies highlighted. Phylogenetic analyses of the morphology of the two sexes recovered three separate species groups within Catharylla: the chelicerata, the mayrabonillae, and the tenellus species groups. The possible position of Micrelephas Schaus, 1922 as sister to Catharylla, based on both morphological and molecular data, and the status of tribe Argyriini are discussed. The biogeographical data indicate that the chelicerata species group is restricted to the Guyanas and the Amazonian regions whereas the tenellus group is restricted to the Atlantic Forest in the South-Eastern part of Brazil. The mayrabonillae group is widespread from Costa Rica to South Bolivia with an allopatric distribution of the two species. COI barcode sequences indicate relatively strong divergence within C. bijuga, C. mayrabonillae, C. serrabonita and C. tenellus.     

The mermithid species Isomermis lairdi (Nematoda,Mermithidae), previously only known in Africa,found in Europe

The present work contributs to the knowledge on the aquatic mermithids (Nematoda, Mermithidae) occurring in black flies – an insufficiently studied group of parasitic nematodes. Isomermis lairdi Mondet, Poinar & Bernadou, 1977, described from larvae of Simulium damnosum Theobald, 1903 in Western Africa, is reported to occur in Bulgaria. The species was isolated from larvae of Simulium ornatum Meigen, 1818 in a local population of simuliids in a mountain stream near Jeleznitsa Village, Sofia district. Postparasitic juveniles of mermithids were obtained from the hosts and reared to the adult stage. The species was identified by morphological and morphometrical characters of postparasitic juveniles, and of male and female individuals. In the summer of 2012 a relatively high rate of mermithid infection in a local host population was detected (prevalence up to 44.1%). In August of the next year host abundance had considerably declined and other simuliid species, Simulium variegatum Meigen, 1818 and Simulium reptans (Linnaeus, 1758), predominated in the investigated locality. In West Africa, Isomermis lairdi is considered to be a potential biological agent for reducing the population density of the Simulium damnosum complex – the main vector of human onchocerciasis. In Europe, species of the Simulium ornatum complex are among the vectors of onchocerciasis of cattle and deer. The mermithids presumably play a certain role in the epidemiology of these diseases. A brief discussion on the taxonomy of the genus Isomermis Coman, 1953, and of the feasibility of molecular methods in mermithid taxonomy is provided. The species Isomermis lairdi is reported for the first time from Europe.     

Superfamily of α-galactosidase MEL genes of the Saccharomyces sensu stricto species complex

In order to study the molecular evolution of the yeasts grouped in the Saccharomyces sensu stricto species complex by analysis of the MEL gene family, we have cloned and sequenced two new species-specific MEL genes from Saccharomyces yeasts: S. paradoxus (MELp) and a Japanese Saccharomyces sp. (MELj). The clones were identified by sequence homology to the S. cerevisiae MEL1 gene. Both clones revealed an ORF of 1413 bp coding for a protein of 471 amino acids. The deduced molecular weights of the α-galactosidase enzymes were 52 767 for MELp and 52 378 for MELj. The nucleotide sequences of the MELp (EMBL accession no. X95505) and the MELj (EMBL accession no. X95506) genes showed 74.7% identity. The degree of identity of MELp to the MEL1 gene was 76.8% and to the S. pastorianus MELx gene, 75.7%. The MELj coding sequence was 75.1% identical to the MEL1 gene and 80.7% to the MELx gene. The data suggest that MEL1, MELj, MELp, and MELx genes are species-specific MEL genes. The strains studied each have only one MEL locus. The MELp gene is located on the S. paradoxus equivalent of S. cerevisiae chromosome X; the MELj gene was on the chromosome that comigrates with the S. cerevisiae chromosome VII/XV doublet and hybridizes to the S. cerevisiae chromosome XV marker HIS3.     

Population dynamics of Frankliniella occidentalis Pergrande and its predator Orius similis Zheng on common crops and surrounding plants

Frankliniella occidentalis Pergrande is important invasive pests in China, causing damage to agricultural production, and Orius similis Zheng is the dominant predator species of F. occidentalis. A two-year survey was conducted to determine the population density of F. occidentalis and O. similis, on chili (Capsicum annuum L.) and maize (Zea mays L.) crops and surrounding weed species, which included white clover (Trifolium repens L.), St. John's wort (Hypericum beanii N. Robson), alfalfa (Medicago sativa L.) and beggarticks (Bidens pilosa L.) in Kunming, southern China. The activity of F. occidentalis on these 6 host plant species was determined using the quartile method. F. occidentalis mainly damaged plants during their flowering stage. The main activity period of F. occidentalis occurred earlier on H. beanii and T. repens than on C. annuum. The peak activity of F. occidentalis occurred in the middle of May (on T. repens). During the whole activity period, the highest thrips densities were recorded on H. beanii among all of the sampled host plant species, followed by C. annuum. The lowest density was recorded on B. pilosa. Dynamics of immature F. occidentalis were more irregular than that of adults. The highest density of O. similis was recorded on Z. mays. It was 2.27–26.43-fold (2017) and 2.01–19.09-fold (2018) higher than that on other host plant species. This study showed that F. occidentalis could migrate between C. annuum and surrounding weeds. The weeds were the main source of thrips on C. annuum. The results indicated that Z. mays can be planted around C. annuum fields as a potential banker plant, to attract O. similis to control F. occidentalis on C. annuum, T. repens and B. pilosa. The flowering period of plants and surrounding plant species has a great effect to the population activities of F. occidentalis and predator O. similis on crops.     

New occurrence of the family Hipponicharionidae (Bradoriida, Arthropoda), in the lower and middle Cambrian of the Cadenas Ibéricas, Spain

The bradoriids Hipponicharion aff. hispanicum and Wimanicharion aff. matthewi are reported from the lower and middle Cambrian strata of the Cadenas Ibéricas, Spain. The genus Hipponicharion seems to be restricted to the Acadobaltic Province. Wimanicharion has been recorded from Sweden and Canada (Nova Scotia). The new discovery of Wimanicharion in Spain indicates its similar palaeobiogeographical distribution to Hipponicharion.     

Differentiation among the Vibrio cholerae serotypes O1, O139, O141 and non-O1, non-O139, non-O141 using specific monoclonal antibodies with dot blotting

Seven different monoclonal antibodies (MAbs) specific to only Vibrio cholerae were produced using a combination of five representative serotypes of V. cholerae for immunization. The first three MAbs (VC-93, VC-82 and VC-223) were specific to the V. cholerae serogroup O1 with different avidity for the serotypes O1 Inaba and O1 Ogawa. The fourth and the fifth MAbs were specific to V. cholerae O139 (VC-812) or O141 (VC-191) serogroups, respectively. The sixth MAb (VC-26) bound to all three serogroups of V. cholerae. The seventh MAb (VC-63) bound to all twenty five isolates of V. cholerae used in this study. None of the seven MAbs showed cross-reactivity with other Vibrio spp. or closely-related V. cholerae species, V. mimicus or other gram-negative bacteria. The eighth MAbs (VC-201) specific to almost all Vibrio spp. was also obtained. In dot blotting, these MAbs can be used to detect a diluted pure culture of V. cholerae in solution with a sensitivity range of from 10⁵ to 10⁷ CFU ml^− 1. However, the detection capability could be improved equivalent to that of PCR technique after preincubation of samples in alkaline peptone water (APW). Thus, these MAbs constitute convenient immunological tools that can be used for simple, rapid and simultaneous direct detection and differentiation of the individual serotypes of V. cholerae in complex samples, such as food and infected animals, without the requirement for bacterial isolation or biochemical characterization.     

Differential secondary metabolite accumulation and performance of Chlosyne lacinia fed with Tithonia diversifolia or Vernonia polyanthes

The larvae of Chlosyne lacinia use Asteraceae species as host plants almost exclusively. The aims of this study were to investigate whether secondary metabolites of Vernonia polyanthes and Tithonia diversifolia are metabolized, excreted intact and/or sequestered during the larval stage of C. lacinia and if they affect feeding behavior or performance. The HPLC-DAD-MS analyses of plants and C. lacinia extracts led to the identification of 25 compounds. Larvae fed with T. diversifolia developed until the 4th instar completing metamorphosis into the adult phase, while larvae fed with V. polyanthes developed only until the 2nd instar. In addition, the larvae in the 3rd and 4th instars fed with T. diversifolia accumulated secondary metabolites taken from these leaves. On the other hand, larvae in the 2nd instar showed an accumulation of apigenin-7-O-glycuronyl and hydroxylated 3-O-E-caffeoylquinic acid when fed with V. polyanthes. The latter compound was probably produced after oxidation of the 3-O-E-caffeoylquinic acid by the phase I metabolism of larvae. Therefore, C. lacinia may have developed tolerance to deterrent compounds produced by T. diversifolia, while some compounds present in V. polyanthes may have been the cause for deficient development and the death of larvae.     

Chromosome numbers in the genus Lippia (Verbenaceae)

The genus Lippia (Verbenaceae) comprises about 200 taxa mainly distributed in Brazil, Mexico, Central America, Africa, Argentina and Paraguay. Some problems involving the number and delimitation of species have been reported. In order to contribute to the solving of these problems, the chromosome numbers of 14 Lippia species are documented. The following species were collected at Espinhaço Range, Southeast Brazil: Section Zapania (L. corymbosa, L. diamantinensis, L. hermannioides, L. lacunosa, L. rotundifolia, L. rubella), section Rhodolippia (L. florida, L. lupulina, L. pseudothea, L. rosella), section Goniostlachyum (L. glandulosa, L. pohliana, L. sidoides) and section Dioicolippia (L.filifolia). Immature inflorescences were collected and the ideal size for chromosome observation was determined. The majority of species have a haploid chromosome number from 10 to 14. Few species have a higher chromosome number, which suggests the occurrence of polyploidy. The relationships between chromosome numbers and the taxonomic sections are also discussed.     

Genetic regulation of flavonoid content in seeds and seedlings of Melilotus alba

Chemical analysis of seeds and seedlings of the CC and cc genotypes in Melilotus alba indicated that these alleles affect flavonoid biosynthesis. The CC seed coats contained orientin and iso-orientin, which were absent in the cc seed coats. The pigment responsible for the red pigmentation of young seedlings of CC genotypes was a cyanidin glycoside. The embryos of seeds of both the CC and cc genotypes contained a flavonoid tentatively identified as a 6,8-di-C-pentosylapigenin. The observation that 3′,4′-dihydroxyflavonoids were absent in the cc genotype and that 4′-hydroxyflavonoids were present in both genotypes indicated that the C/c alleles controlled the 3′-hydroxylation of flavonoids. The C/c alleles did not, however, control 3′-hydroxylation of cinnamic acids since caffeic acid was detected in both genotypes.     

Cloning of the modification methylase gene of Bacillus centrosporus in Escherichia coli

The gene specifying a sequence-specific modification methylase of Bacillus centrosporus has been cloned in Escherichia coli using the restriction endonuclease HindIII and the plasmid pBR322. The selection was based on detection of new methylation properties rendering recombinant plasmids carrying the methylase gene nonsusceptible to BcnI endonuclease cleavage. The presence of a 3.2-kb HindIII fragment in either orientation conferred BcnI resistance on the recombinant plasmids. These results suggest that the BcnI methylase gene is expressed in E. coli under the control of a promoter located on the cloned fragment. The relative level of BcnI methylase enzyme in E. coli was similar to that in B. centrosporus. The recombinant clones do not exhibit any BcnI restriction-endonuclease activity.