Investigating species boundaries using DNA and morphology in the mite <Emphasis Type="Italic">Tyrophagus curvipenis</Emphasis> (Acari: Acaridae), an emerging invasive pest,with a molecular phylogeny of the genus <Emphasis Type="Italic">Tyrophagus</Emphasis>

Mites of the genus Tyrophagus (Acari: Acaridae) are among the most widespread and common mites, inhabiting diverse natural and anthropogenic habitats. Some species are pests of agricultural products and stored food and/or live in house dust, causing allergies to humans. We sequenced 1.2 kb of the mitochondrial COI gene for 38 individuals belonging to seven species of Tyrophagus, including T. curvipenis, T. putrescentiae, T. fanetzhangorum, T. longior, T. perniciosus, and T. cf. similis. Molecular phylogenetic analyses (1) recovered two major clades corresponding to the presence or absence of eyespots, and (2) separated all included morphological species. Tyrophagus curvipenis and T. putrescentiae had the lowest between-species genetic distances (range, mean?±?SD): 14.20–16.30, 15.17?±?0.40 (K2P). The highest within-species variation was found in T. putrescentiae 0.00–4.33, 1.78?±?1.44 (K2P). In this species, we recovered two distinct groups; however, no geographical or ecological dissimilarities were observed between them. Based on our analyses, we document important morphological differences between T. curvipenis and T. putrescentiae. For the first time, we record the occurrence of T. curvipenis in the New World and suggest that it may be an emerging pest as it is currently spreading in agricultural produce.     

Unreduced gamete formation in wheat × <Emphasis Type="Italic">Aegilops</Emphasis> spp. hybrids is genotype specific and prevented by shared homologous subgenomes

Key message

The presence of homologous subgenomes inhibited unreduced gamete formation in wheat × Aegilops interspecific hybrids. Unreduced gamete rates were under the control of the wheat nuclear genome.

Abstract

Production of unreduced gametes is common among interspecific hybrids, and may be affected by parental genotypes and genomic similarity. In the present study, five cultivars of Triticum aestivum and two tetraploid Aegilops species (i.e. Ae. triuncialis and Ae. cylindrica) were reciprocally crossed to produce 20 interspecific hybrid combinations. These hybrids comprised two different types: T. aestivum × Aegilops triuncialis; 2n = ABDU^tC^t (which lack a common subgenome) and T. aestivum × Ae. cylindrica; 2n = ABDD^cC^c (which share a common subgenome). The frequency of unreduced gametes in F₁ hybrids was estimated in sporads from the frequency of dyads, and the frequency of viable pollen, germinated pollen and seed set were recorded. Different meiotic abnormalities recorded in the hybrids included precocious chromosome migration to the poles at metaphase I and II, laggards in anaphase I and II, micronuclei and chromosome stickiness, failure in cell wall formation, premature cytokinesis and microspore fusion. The mean frequency of restitution meiosis was 10.1 %, and the mean frequency of unreduced viable pollen was 4.84 % in T. aestivum × Ae. triuncialis hybrids. By contrast, in T. aestivum × Ae. cylindrica hybrids no meiotic restitution was observed, and a low rate of viable gametes (0.3 %) was recorded. This study present evidence that high levels of homologous pairing between the D and D^c subgenomes may interfere with meiotic restitution and the formation of unreduced gametes. Variation in unreduced gamete production was also observed between T. aestivum × Ae. triuncialis hybrid plants, suggesting genetic control of this trait.

     

Early characterization of somatic hybrids from symmetric protoplast electrofusion of <Emphasis Type="Italic">Solanum pinnatisectum</Emphasis> Dun. and <Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.

Variability of 31 somatic hybrids of Solanum pinnatisectum Dun. with Solanum tuberosum L. for leaf morphology, plant vigor, resistance to Phytophthora infestans, ploidy level, and cytoplasm type was evaluated in vitro. The composition of these somatic hybrids was as follows: [S. pinnatisectum Dun. (2n = 2x = 24; cytoplasmic type Wγ) + S. tuberosum L. (2n = 4x = 48; cytoplasmic type Tß)]. Based on leaf morphology and plant growth vigor, plants were divided into three groups, including plants close to tbr parent with unlobed leaves, small plants with scarcely dissected leaves, and vigorous plants with asymmetrically and pinnately lobed leaves. Nine of the somatic hybrids were found to be highly resistant to P. infestans. Somatic hybrids were either tetraploid or hexaploid, with hexaploids being predominant. The cytoplasm of somatic hybrids was either Tßγ or Wßγ, with Tßγ being more common. Overall, in contrast to leaf morphology and growth vigor, level of resistance to P. infestans was not related to either ploidy level or type of cytoplasm. These findings demonstrate that early in vitro selection of promising hybrids can be useful in breeding programs.     

Maternal care,larviparous and oviparous reproduction of <Emphasis Type="Italic">Hypoaspis larvicolus</Emphasis> (Acari: Laelapidae) feeding on astigmatid mites

Hypoaspis larvicolus (Acari: Laelapidae) (first report from Turkey) occurred together with Sancassania polyphyllae (Acari: Acaridae) on the larvae of the scarab beetle, Polyphylla fullo (Coleoptera: Scarabaeidae), that were feeding on the roots of strawberry in Aydin, Turkey. Laboratory studies were conducted to (1) observe whether H. larvicolus feeds and completes its life cycle on the various stages of S. polyphyllae or other astigmatid mites, such as Acarus siro, Carpoglyphus lactis and Tyrophagus putrescentiae (Acaridae), and to determine its population growth when feeding on these prey, and (2) to determine development periods, longevity and fecundity of H. larvicolus feeding on C. lactis. Hypoaspis larvicolus females did not feed on S. polyphyllae, but fed, developed and reproduced when A. siro, C. lactis or T. putrescentiae were provided as prey. Hypoaspis larvicolus is larviparous as well as oviparous. The female lays eggs or gives birth to larvae. If a female gives birth to a larva, it is attached under the female’s venter for 1–2 days, a phenomenon recorded for the first time in Hypoaspis; in fact, for the first time in mites. The results of the population growth experiments revealed that H. larvicolus feeding on C. lactis produced the highest number of eggs, juveniles and adults. The developmental periods of H. larvicolus feeding on C. lactis at life-cycle path I (larva to adult) and II (egg to adult) were 12.2?±?0.3 and 15.6?±?0.6 days (females) and 19.5?±?0.2 and 20.9?±?0.4 days (males), respectively. Longevity of females versus males of H. larvicolus was 120.6?±?7.2 versus 91.6?±?13.1 days (life cycle I) and 110.0?±?27.7 versus 118.3?±?10.9 days (life cycle II), respectively.     

Comparison of Killing Activity of Micafungin Against Six <Emphasis Type="Italic">Candida</Emphasis> Species Isolated from Peritoneal and Pleural Cavities in RPMI-1640, 10 and 30% Serum

Currently echinocandins are recommended in Candida peritonitis and pleuritis. We determined micafungin killing rates (k values) at therapeutic concentrations (0.25–2 mg/L) in RPMI-1640 with and without 10 and 30% serum mimicking in vivo conditions against six Candida species isolated from peritoneal and pleural fluid. In RPMI-1640, micafungin was fungicidal against C. glabrata, C. krusei and C. kefyr within 2.27?±?10.68, 2.69?±?10.29 and 3.10?±?4.41 h, respectively, while was fungistatic against C. albicans, C. tropicalis and C. parapsilosis. In 10% serum, ≥?0.25, ≥?0.5, ≥?0.5 and ≥?1 mg/L micafungin produced positive k values (killing) for all C. albicans, C. glabrata, C. kefyr and C. krusei, respectively. In 30% serum, 2 mg/L micafungin produced killing against all C. albicans, C. glabrata and C. kefyr isolates, but was ineffective against C. krusei, C. parapsilosis and 2 of 3 C. tropicalis. Micafungin exposure should be increased against non-albicans species to eradicate fungi from peritoneal and pleural cavities.     

Stability of fumonisin B<Subscript>1</Subscript>, deoxynivalenol,zearalenone, and T-2 toxin during processing of traditional Nigerian beer and spices

The stability of the Fusarium mycotoxins fumonisin B₁, deoxynivalenol, T-2 toxin, and zearalenone during processing of Nigerian traditional spices (dawadawa, okpehe, and ogiri) and beer (burukutu) using artificially contaminated raw materials was investigated. Results revealed the reduction of these toxins in all the final products. Boiling played a significant role (p?<?0.05) in Fusarium mycotoxin reduction in the traditional spices. The highest percentage reduction of deoxynivalenol (76%) and zearalenone (74%) was observed during okpehe processing (boiled for 12 h). Dehulling and fermentation further demonstrated a positive influence on the reduction of these toxins with a total reduction ranging from 85 to 98% for dawadawa, 86 to 100% for okpehe, and 57 to 81% for ogiri. This trend was also observed during the production of traditional beer (burukutu), with malting and brewing playing a major impact in observed reduction. In addition, other metabolites including deoxynivalenol-3-glucoside, 15-acetyl-deoxynivalenol, α-zearalenol, and β-zearalenol which were initially not present in the raw sorghum were detected in the final beer product at the following concentrations 26?±?11, 16?±?7.7, 22?±?18, and 31?±?16 μg/kg, respectively. HT-2 toxin was also detected at a concentration of 36?±?13 μg/kg along the processing chain (milled malted fraction) of the traditional beer. For the traditional spices, HT-2 toxin was detected (12 μg/kg) in ogiri. Although there was a reduction of mycotoxins during processing, appreciable concentrations of these toxins were still detected in the final products. Thus, the use of good quality raw materials significantly reduces mycotoxin contamination in final products.     

A multiplexed set of microsatellite markers for discriminating <Emphasis Type="Italic">Acacia mangium</Emphasis>, <Emphasis Type="Italic">A. auriculiformis</Emphasis>, and their hybrid

In order to assist breeding and gene pool conservation in tropical Acacias, we aimed to develop a set of multipurpose SSR markers for use in both Acacia mangium and A. auriculiformis. A total of 51 SSR markers (developed in A. mangium and natural A. mangium x A. auriculiformis hybrid) were tested. A final set of 16 well-performing SSR markers were identified, six of which were species diagnostic. The markers were optimized for assay in four multiplex mixes and used to genotype range-wide samples of A. mangium, A. auriculiformis, and putative F₁ hybrids. Simulation analysis was used to investigate the power of the markers for identifying the pure species and their F₁, F₂, and backcross hybrids. The six species diagnostic markers were particularly powerful for detecting F₁ hybrids from pure species but could also discriminate the pure species from F₂ and backcross progenies in most cases (97 %). STRUCTURE analysis using all 16 markers was likewise able to distinguish these cross types and pure species sets. Both sets of markers had difficulties in distinguishing F₂ and backcross progenies. However, identifying F₁ from pure species is the current primary concern in countries where these species are planted. The SSR marker set also has direct application in DNA profiling (probability of identity?=?4.1?×?10^?13), breeding system analysis, and population genetics.     

Development and reproduction of <Emphasis Type="Italic">Amblyseius largoensis</Emphasis> (Acari: Phytoseiidae) feeding on pollen, <Emphasis Type="Italic">Raoiella indica</Emphasis> (Acari: Tenuipalpidae), and other microarthropods inhabiting coconuts in Florida,USA

The red palm mite, Raoiella indica (Acari: Tenuipalpidae), is an important pest of palms (Arecaceae) and other species within the Zingiberaceae, Musaceae and Strelitziaceae families. Raoiella indica was discovered in the USA (Palm Beach and Broward counties, Florida) late in 2007, and it subsequently spread to other Florida counties. The predatory mite Amblyseius largoensis (Acari: Phytoseiidae) has been found associated with R. indica in Florida. In order to verify whether A. largoensis can develop and reproduce when feeding exclusively on R. indica, the biology of this predator was evaluated on various food sources, including R. indica. Five diets [R. indica, Tetranychus gloveri¸ Aonidiella orientalis, Nipaecocus nipae, oak (Quercus virginiana) pollen] and a no-food control were tested to determine the predators’ development, survivorship, oviposition rate, sex ratio and longevity at 26.5 ± 1°C, 70 ± 5% RH and a 12:12 L:D photophase. Amblyseius largoensis was able to complete its life cycle and reproduce when fed exclusively on R. indica. The development of immature stages of A. largoensis was faster and fecundity and survivorship were higher when fed on R. indica or T. gloveri compared to the other food sources. The intrinsic rate of natural increase of A. largoensis was significantly higher when fed on R. indica than on other diets. These results suggest that, despite earlier assessments, A. largoensis can play a role in controlling R. indica.     

<Emphasis Type="Italic">Talaromyces heiheensis</Emphasis> and <Emphasis Type="Italic">T. mangshanicus</Emphasis>, two new species from China

Two new species, Talaromyces heiheensis from rotten wood and T. mangshanicus isolated from soil, are illustrated and described as new to science in sections Trachyspermi and Talaromyces. The phylogenetic positions of the two new species inferred from the internal transcribed spacer, beta-tubulin, calmodulin and RNA polymerase II second largest subunit regions were carried out. Talaromyces heiheensis is phylogenetically closely related to T. albobiverticillius, T. rubrifaciens, T. solicola and T. erythromellis, and characterised by slow growth on Czapek yeast autolysate agar at 25 °C, orange conidia en masse on malt extract agar at 25 °C, biverticillate and terverticillate conidiophores, acerose phialides and subglobose to ellipsoidal, smooth-walled conidia. Talaromyces mangshanicus is related to T. kendrickii, T. qii and T. thailandensis, and characterised by slow-growing colonies with absent or sparse sporulation on CYA agar at 25 °C, conidia en masse greyish purple, purplish red soluble pigment on yeast extract agar (YES) at 25 °C, biverticillate conidiophores, ampulliform phialides and subglobose to ellipsoidal conidia with echinulate walls. They are distinguished from the known species in culture characteristics on four standard media, microscopic features and sequence data.     

Efficiency of spinosad, <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> and <Emphasis Type="Italic">Trichogramma brassicae</Emphasis> against the tomato leafminer in greenhouse

The objective of this study was to evaluate the efficacy of three eco-friendly control agents, either singly or in a pairwise combination, for the control of the tomato leafminer, Tuta absoluta (Meyrick) (Lep: Gelechiidae). They include the naturally derived pesticide spinosad, a commercially available formulation of Bacillus thuringiensis var. Kurstaki (Bt), and a native population of Trichogramma brassicae Bezdenko (Hym: Trichogrammatidae). Tomato plants containing the T. absoluta were treated with one of the seven following treatments in a greenhouse: (1) a single release of T. brassicae against the eggs; (2) two applications of Bt (2 kg ha^?1); (3) and (4) one application of spinosad at two rates (60 and 120 g a.i. ha^?1); (5) T. brassicae release?+?Bt spray; (6) T. brassicae release?+?spinosad spray; and (7) spinosad spray?+?Bt spray. The highest mortality rate was recorded for the spinosad?+?Bt (88.33?±?1.43%) and T. brassicae?+?spinosad (78.33?±?3.74%) combinations, respectively; while the lowest mortality rate was obtained through the single application of T. brassicae (31.67?±?4.84%). Based on our results, the Bt and spinosad seem to be suitable candidates for combination with other biological and cultural techniques towards an integrated management of the tomato leafminer.     

Rediscovery of type material of the bioerosional trace fossil <Emphasis Type="Italic">Talpina</Emphasis> von Hagenow, 1840 and its ichnotaxonomic implications

Friedrich von Hagenow is renowned for his seminal inventory of the diverse invertebrate fauna from the lower Maastrichtian chalk (Upper Cretaceous) of the Isle of Rügen, Germany. Tragically, his immense collection at Stettin (today Szczecin, Poland) was almost completely destroyed during World War II, including the type material of hundreds of species established by him during the first half of the 19th century. Seven belemnite specimens from his collection rediscovered at the Museum für Naturkunde Berlin, as well as 14 belemnites traced at the Museum für Mineralogie und Geologie at the Senckenberg Naturhistorische Sammlungen Dresden, include syntypes of four Talpina ichnospecies, labelled T. ramosa, T. solitaria, T. foliacea and T. sentiformis. To document this material, a new ‘digital cylinder seal technique’ was applied. The rediscovered material allows lectotype designations for T. ramosa and T. solitaria (now Trypanites solitarius), whereas T. foliacea and T. sentiformis, which where not formally published by von Hagenow, are regarded as nomina nuda. As a consequence, the non-topotypical neotypes of T. ramosa and T. solitaria, as designated by Ehrhard Voigt in 1972, are obsolete.     

Chemical and molecular identification of the invasive termite <Emphasis Type="Italic">Zootermopsis nevadensis</Emphasis> (Isoptera: Archotermopsidae) in Japan

Numerous termite species have been introduced outside their native ranges by human transport, and some have become invasive. The dampwood termite Zootermopsis nevadensis (Hagen), which is native to western North America, has been introduced to and become established in Kawanishi City, Hyogo Prefecture, Japan. Zootermopsis nevadensis is subdivided into two subspecies based on cuticular hydrocarbon (CHC) phenotypes: Z. nevadensis nevadensis and Z. nevadensis nuttingi (Haverty and Thorne). Here, we identified Z. nevadensis in Japan as hybrids between the two subspecies. Chemical analysis showed the presence of 7,15-dimethylhenicosane and 5,17-dimethylhenicosane in the CHCs of Z. nevadensis in Japan, corresponding to the CHC phenotype of Z. n. nevadensis. Conversely, all mitochondrial cytochrome c oxidase subunit I sequences of Z. nevadensis in Japan were identical to sequences from Z. n. nuttingi and hybrids between the two subspecies from a native hybrid zone in California, USA. In addition, phylogenetic analysis showed that Z. nevadensis in Japan formed a clade with Z. n. nuttingi and hybrids between the two subspecies. Our results show discordance between the chemical and genetic features of Z. nevadensis in Japan, indicating that individuals of Z. nevadensis in Japan are hybrids between the two subspecies.     

Estimates of Heritability for Growth and Shell Color Traits and Their Genetic Correlations in the Black Shell Strain of Pacific Oyster <Emphasis Type="Italic">Crassostrea gigas</Emphasis>

The Pacific oyster Crassostrea gigas has been introduced widely and massively and became an economically important aquaculture species on a global scale. We estimated heritabilities of growth and shell color traits and their genetic correlations in black shell strain of C. gigas. Analyses were performed on 22 full-sib families in a nested mating design including 410 individuals at harvest (24 months of age). The parentage assignment was inferred based on four panels of multiplex PCR markers including 10 microsatellite loci and 94.9% of the offspring were unambiguously assigned to single parent pairs. The Spearman correlation test (r = ? 0.992, P < 0.001) demonstrated the high consistency of the shell pigmentation (SP) and L* and their same efficacy in shell color measurements. The narrow-sense heritability estimated under the animal model analysis was 0.18 ± 0.12 for shell height, 0.25 ± 0.16 for shell length, 0.10 ± 0.09 for shell width, 0.42 ± 0.20 for total weight, 0.32 ± 0.18 for shell weight, and 0.68 ± 0.16 for L*, 0.69 ± 0.16 for shell pigmentation, respectively. The considerable additive genetic variation in growth and shell color traits will make it feasible to produce genetic improvements for these traits in selective breeding program. High genetic and phenotypic correlations were found among growth traits and among shell color traits. To optimize a selection strategy for both fast growth and pure dark shell strain of C. gigas, it is proposed to take both total weight and black shell as joint objective traits in selective breeding program. Our study offers an important reference in the process of selective breeding in black shell color stain of C. gigas and will facilitate to develop favorable breeding strategies of genetic improvements for this economically important strain.     

Using SSR markers for hybrid identification and resource management in Vietnamese <Emphasis Type="Italic">Acacia</Emphasis> breeding programs

We used a set of 16 SSR markers to check the identity of pure-species and hybrid clones in Vietnam’s Acacia auriculiformis, Acacia mangium, and acacia hybrid (A. mangium × A. auriculiformis) breeding programs. The statistics package HIest, applied to a large synthesized population, enabled accurate allocation of genotypes to the two pure species, F₁ and F₂ inter-specific hybrids and backcrosses, based on estimates of hybridity and heterozygosity. The hybridity status of putatively pure A. mangium and A. auriculiformis clones in adjacent clonal seed orchards was checked. Four out of 100 clones selected as A. mangium were found to be backcrosses (A. mangium × F₁ inter-specific hybrid) while out of 96 clones selected as A. auriculiformis, two were F₁ hybrids and two were backcrosses (A. auriculiformis × F₁ hybrid). The markers were then applied to check the hybridity status of 160 putative acacia F₁ hybrid genotypes that had been selected on morphological criteria from open-pollinated progenies collected from A. auriculiformis and A. mangium parents. Many selections based on morphology were found to be mistaken. Only thirteen of 63 clones originating from A. auriculiformis mothers were F₁ hybrids, four were backcrosses, and the remaining 46 were pure A. auriculiformis. Fewer mistakes were evident for clones selected from A. mangium mothers, with 82 out of 89 clones confirmed as F₁ hybrids, three as backcrosses, and four as pure A. mangium. The occurrence of F₁ hybrids and backcrosses in pure-species seed orchards and their progeny shows that inter-species contamination is an issue requiring management in both pure-species and in hybrid breeding of these species in Vietnam. Examination of genetic distances among verified clones showed patterns of relatedness that were consistent with pedigree records. Implications for resource management as well as for breeding and clonal selection strategies are considered.     

<Emphasis Type="Italic">Tamarix</Emphasis> (Tamaricaceae) hybrids: the dominant invasive genotype in southern Africa

Hybridization is regarded as a rapid mechanism for increasing genetic variation that can potentially enhance invasiveness. Tamarix hybrids appear to be the dominant genotypes in their invasions. Exotic Tamarix are declared invasive in South Africa and the exotic T. chinensis and T. ramosissima are known to hybridize between themselves, and with the indigenous T. usneoides. However, until now, it was not known which species or hybrid is the most prevalent in the invasion. With a biocontrol programme being considered as a way of suppressing the alien Tamarix populations, it is important to document the population genetic dynamics of all species in the region. This investigation sought to identify Tamarix species in southern Africa and their hybrids, describe their population structure, and reveal the geographic origin of the invasive species. To achieve this, nuclear Internal Transcribed Spacer (ITS) sequence data and the multilocus Amplified Fragment Length Polymorphisms (AFLPs) markers were used. Phylogenetic analysis and population genetic structure confirmed the presence of three species in South Africa (T. chinensis, T. ramosissima and T. usneoides) with their hybrids. The indigenous T. usneoides is clearly genetically distant from the alien species T. chinensis and T. ramosissima. Interestingly, the Tamarix infestation in South Africa is dominated (64.7 %) by hybrids between T. chinensis and T. ramosissima. The exotic species match their counterparts from their places of origin in Eurasia, as well as those forming part of the invasion in the US.     

Suitability of edaphic arthropods as prey for <Emphasis Type="Italic">Proctolaelaps bickleyi</Emphasis> and <Emphasis Type="Italic">Cosmolaelaps brevistilis</Emphasis> (Acari: Mesostigmata: Melicharidae,Laelapidae) under laboratory conditions

Soils are often complex habitats inhabited by a wide range of organisms, some harmful to plants and others beneficial, for example by attacking harmful organisms. Beneficial organisms include predatory mites, some of which have been commercialized for biological control of pest insects and mites. The objective of this work was to evaluate under laboratory condition the suitability of representative soil insect and mite pests, especially Aceria tulipae (Keifer), as prey to the soil-inhabiting predatory mites Proctolaelaps bickleyi (Bram) and Cosmolaelaps brevistilis (Karg). Predation, oviposition and survivorship of recently molted adult females of the predators were assessed in the dark in rearing chambers at 25 ± 1 °C and 75 ± 3% RH. Predation rate by P. bickleyi on A. tulipae was significantly higher than that by C. brevistilis (196.3 vs. 71.0 specimens/day). About 482 A. tulipae were preyed by each P. bickleyi at each day, when 500 A. tulipae were made available daily to the predator. Oviposition rate on that prey was also higher for P. bickleyi (4.2 eggs/day). For C. brevistilis, the highest level of oviposition was on Caliothrips phaseoli (Hood) (1.2 eggs/day). Survivorship was always higher for C. brevistilis (≥ 70%), given its ability to remain alive relatively long even in the absence of prey. High rates of survivorship of P. bickleyi were observed on A. tulipae, Bradysia matogrossensis (Lane) and Protorhabditis sp. Promising results were obtained for P. bickleyi on A. tulipae and even on other prey, justifying the conduction of complementary studies under field condition.     

Human milk and mucosal lacto- and galacto-<Emphasis Type="Italic">N</Emphasis>-biose synthesis by transgalactosylation and their prebiotic potential in <Emphasis Type="Italic">Lactobacillus</Emphasis> species

Lacto-N-biose (LNB) and galacto-N-biose (GNB) are major building blocks of free oligosaccharides and glycan moieties of glyco-complexes present in human milk and gastrointestinal mucosa. We have previously characterized the phospho-β-galactosidase GnbG from Lactobacillus casei BL23 that is involved in the metabolism of LNB and GNB. GnbG has been used here in transglycosylation reactions, and it showed the production of LNB and GNB with N-acetylglucosamine and N-acetylgalactosamine as acceptors, respectively. The reaction kinetics demonstrated that GnbG can convert 69 ± 4 and 71 ± 1 % of o-nitrophenyl-β-d-galactopyranoside into LNB and GNB, respectively. Those reactions were performed in a semi-preparative scale, and the synthesized disaccharides were purified. The maximum yield obtained for LNB was 10.7 ± 0.2 g/l and for GNB was 10.8 ± 0.3 g/l. NMR spectroscopy confirmed the molecular structures of both carbohydrates and the absence of reaction byproducts, which also supports that GnbG is specific for β1,3-glycosidic linkages. The purified sugars were subsequently tested for their potential prebiotic properties using Lactobacillus species. The results showed that LNB and GNB were fermented by the tested strains of L. casei, Lactobacillus rhamnosus (except L. rhamnosus strain ATCC 53103), Lactobacillus zeae, Lactobacillus gasseri, and Lactobacillus johnsonii. DNA hybridization experiments suggested that the metabolism of those disaccharides in 9 out of 10 L. casei strains, all L. rhamnosus strains and all L. zeae strains tested relies upon a phospho-β-galactosidase homologous to GnbG. The results presented here support the putative role of human milk oligosaccharides for selective enrichment of beneficial intestinal microbiota in breast-fed infants.     

Productivity and biochemical composition of <Emphasis Type="Italic">Tetradesmus obliquus</Emphasis> and <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis>: effects of different cultivation approaches

The present work evaluated biomass productivity, carbon dioxide fixation rate, and biochemical composition of two microalgal species, Phaeodactylum tricornutum (Bacillariophyta) and Tetradesmus obliquus (Chlorophyta), cultivated indoors in high-technology photobioreactors (HT-PBR) and outdoors both in pilot ponds and low-technology photobioreactors in a greenhouse in southern Italy. Microalgae were grown in standard media, under nitrogen starvation, and in two liquid digestates obtained from anaerobic digestion of agro-zootechnical and vegetable biomass. P. tricornutum, cultivated in semi-continuous mode in indoor HT-PBRs with standard medium, showed a biomass productivity of 21.0?±?2.3 g m^?2 d^?1. Applying nitrogen starvation, the lipid productivity increased from 2.3 up to 4.5?±?0.5 g m^?2 d^?1, with a 24 % decrease of biomass productivity. For T. obliquus, a biomass productivity of 9.1?±?0.9 g m^?2 d^?1 in indoor HT-PBR was obtained using standard medium. Applying liquid digestates as fertilizers in open ponds, T. obliquus gave a biomass productivity (10.8?±?2.0 g m^?2 d^?1) not statistically different from complete medium such as P. tricornutum (6.5?±?2.2 g m^?2 d^?1). The biochemical data showed that the fatty acid composition of the microalgal biomass was affected by the different cultivation conditions for both microalgae. In conclusion, it was found that the microalgal productivity in standard medium was about doubled in HT-PBR compared to open ponds for P. tricornutum and was about 20 % higher for T. obliquus.     

<Emphasis Type="Italic">Nibribacter flagellatus</Emphasis> sp. nov., isolated from rhizosphere of <Emphasis Type="Italic">Hibiscus syriacus</Emphasis> and emended description of the genus <Emphasis Type="Italic">Nibribacter</Emphasis>

A Gram-stain negative, aerobic, motile by flagella, rod-shaped strain (THG-T16^T) was isolated from rhizosphere of Hibiscus syriacus. Growth occurred at 10–40 °C (optimum 28–30 °C), at pH 6.0–8.0 (optimum 7.0) and at 0–1.0% NaCl (optimum 0%). Based on 16S rRNA gene sequence analysis, the near phylogenetic neighbours of strain THG-T16^T were identified as Nibribacter koreensis KACC 16450^T (98.6%), Rufibacter roseus KCTC 42217^T (94.7%), Rufibacter immobilis CCTCC AB 2013351^T (94.5%) and Rufibacter tibetensis CCTCC AB 208084^T (94.4%). The DNA G+C content of strain THG-T16^T was determined to be 46.7 mol%. DNA–DNA hybridization values between strain THG-T16^T and N. koreensis KACC 16450^T, R. roseus KCTC 42217^T, R. immobilis CCTCC AB 2013351^T, R.tibetensis CCTCC AB 208084^T were 33.5?±?0.5% (31.7?±?0.7% reciprocal analysis), 28.1?±?0.2% (25.2?±?0.2%), 17.1?±?0.9% (10.2?±?0.6%) and 8.1?±?0.3% (5.2?±?0.1%). The polar lipids were identified as phosphatidylethanolamine, two unidentified aminophospholipids, an unidentified aminolipid and three unidentified lipids. The quinone was identified as MK-7 and the polyamine as sym-homospermidine. The major fatty acids were identified as C_16:1 ω5c, C_17:1 ω6c, iso-C_15:0, summed feature 3 (C_16:1 ω7c and/or C_16:1 ω6c) and summed feature 4 (iso-C_17:1 I and/or anteiso-C_17:1 B). On the basis of the phylogenetic analysis, chemotaxonomic data, physiological characteristics, and DNA–DNA hybridization data, strain THG-T16^T represents a novel species of the genus Nibribacter, for which the name Nibribacter flagellatus sp. nov. is proposed. The type strain is THG-T16^T(=?KACC 19188^T?=?CCTCC AB 2016246^T).     

Hybridization success is largely limited to homoploid <Emphasis Type="Italic">Prunus</Emphasis> hybrids: a multidisciplinary approach

Prunus fruticosa is a rare shrub occurring in Eurasian thermophilous forest-steppe alliances. The species frequently hybridizes with cultivated Prunus species in Europe (allochthonous tetraploid P. cerasus and partly indigenous diploid P. avium). Propidium iodide flow cytometry, distance-based morphometrics, elliptic Fourier analysis and embryology were employed to evaluate the extent of hybridization in six Slovak populations. Flow cytometric analyses revealed three ploidy levels: diploid (P. avium), triploid (P. × mohacsyana) and tetraploid (P. fruticosa, P. × eminens and P. cerasus). In addition, P. fruticosa and P. cerasus, at the tetraploid level, were found to differ in absolute genome size. An embryological evaluation suggested the existence of a triploid block in P. × mohacsyana and significant potential for hybridization among tetraploid taxa (indicated also by a continuous distribution of genome size data and further mirrored by morphometrics). Although hybrids significantly differ in ploidy level and embryological characteristics, they are almost indistinguishable using morphological characters. Hybridization with P. cerasus thus turns out to be a significant threat to wild populations of P. fruticosa compared to the relatively weak influence of P. avium.