Hidden diversity uncovered in Hygrophorus sect. Aurei (Hygrophoraceae), including the Mediterranean H. meridionalis and the North American H. boyeri, spp. nov.

For many years, the binomial Hygrophorus hypothejus was widely applied to collections from various geographical regions in different continents, assuming a circum-boreal and circum-mediterranean distribution for this species. This hypothesis, however, had never been put to the test. To assess the diversity and species-limits within this complex of yellow-coloured waxcaps, a phylogenetic, morphological and taxonomical investigation into Hygrophorus sect. Aurei and similar species in sect. Olivaceoumbrini was carried out, including material of pan-European origin, as well as the east and west coasts of North America. Following sequencing of the ITS rDNA locus, nine lineages are confirmed in sect. Aurei, most of them highly continentalised. Of these, two are new to science, introduced here as Hygrophorus boyeri sp. nov., from Pinus banksiana and P. rigida forests in eastern North America and from P. muricata and P. contorta forests in western North America, and Hygrophorus meridionalis sp. nov., from Pinus brutia and Pinus halepensis forests in the island of Cyprus and mainland Greece. H. hypothejus is lectotypified and epitypified, and here resolved as a strictly European species, with the old forgotten taxon Hygrophorus siccipes revived as its North American vicariant. The placement of Hygrophorus fuligineus in sect. Aurei is phylogenetically confirmed and detailed comparisons between morphologically similar and phylogenetically affiliated taxa in sect. Aurei and sect. Olivaceoumbrini are provided. The chronic confusion associated with Hygrophorus fuscoalbus, a highly controversial taxon described from Germany nearly two centuries ago and variously interpreted since, is discussed, concluding that this name is too ambiguous to be applied to any currently recognized species.     

The life cycle of Hymenoscyphus fraxineus on Manchurian ash,Fraxinus mandshurica,in Japan

Hymenoscyphus fraxineus causes a lethal disease known as “ash dieback” in the common ash, Fraxinus excelsior, in Europe. It is hypothesized that the fungus originated from East Asia. This fungus is found on the leaf litter of the Manchurian ash, Fraxinus mandshurica, in Japan and is reported to produce apothecia on pseudosclerotial plates formed mainly on decomposing rachises. However, dieback disease has not been reported in Japan, and little is known about the life cycle of H. fraxineus. This study was conducted to explore the behavior and life cycle of this fungus. It was revealed that, after infection by ascospores, H. fraxineus endophytically inhabits the living leaves of F. mandshurica. On fallen leaves, the fungus behaves saprophytically, producing apothecia on pseudosclerotial plates formed mainly on the decomposing rachises. Analysis by real-time quantitative polymerase chain reaction (qPCR) revealed that the amount of H. fraxineus DNA sharply increased in rachises, while such sharp increase of DNA was not found in leaflets.     

Population genetic structure in a threatened tree, Pyrus calleryana var. dimorphophylla revealed by chloroplast DNA and nuclear SSR locus polymorphisms

Pyrus calleryana var. dimorphophylla, a variety of Callery pear (Pyrus calleryana), is endemic to the Tokai district of central Japan, and is currently listed as “Endangered”. The remnant habitats and trees are of limited number, and highly fragmented. As the first step in determining appropriate conservation units, genetic diversity and differentiation in this species were investigated using chloroplast DNA (cpDNA) and nuclear simple sequence repeat (SSR) polymorphisms. All possible remnant trees were genotyped, then six populations were defined based on the results of cpDNA haplotype determination and Bayesian clustering approaches performed using the SSR locus data. Some trees appeared to originate from artificial propagation. Some individuals were difficult to differentiate genetically from the related species, Pyrus × uyematsuana, which is considered to be a hybrid between P. calleryana var. dimorphophylla and a possibly naturalized species, Pyrus pyrifolia, implying that introgression between these species may have occurred. In P. calleryana var. dimorphophylla, anthropogenic factors such as propagation and related species planting are probably major causes of complexity in the genetic structure.     

Hereditäre Paragangliome

Hereditary paragangliomas/pheochromocytomas are inherited as autosomal dominant traits. Three types, i. e. PGL1, PGL2, and PGL3 can be distinguished. They are caused by mutations of the genes SDHD, SDHC, and SDHB, which encode components of mitochondrial complex II (succinate-ubiquinone reductase, SDH) of the respiratory chain. “Loss of heterozygosity” (LOH) of the region harbouring the disease gene is found in all 3 types of paragangliomas. LOH results in functional loss of SDH, the accumulation of succinate and of reactive oxygen species. As a consequence, hypoxia-dependent metabolic pathways are induced which appear to trigger tumorigenesis. PGL3 and PGL4 can be caused by germ-line mutations in either the paternal or the maternal copy of the respective disease gene, i. e. SDHC and SDHB. In contrast, PGL1 only occurs after paternal transmission of a mutation in SDHD. This observation can be explained by partial inactivation (“imprinting”) of the maternal SDHD-gene and induction of hypoxia-dependent genes, which in turn favour non-disjunction and loss of chromosome 11.     

On the status of “Urotherium antiquum” (Ameghino) (Xenarthra,Glyptodontidae)

Within the Late Neogene Glyptodontidae of the Pampean region of Argentina, “Urotherium antiquum” was described on the basis of some associated osteoderms of the dorsal carapace, which in addition include a partial skull and left hemimandible. The diagnostic characters are located on the exposed surface of the osteoderms of the dorsal carapace which somewhat resembles those of the Pleistocene genus Neuryurus. Although the relationship of “U. antiquum” to the remaining Glyptodontidae has never been clarified, some cladistic analyses suggest a close phylogenetic affinity with the clade composed of Plohophorus + (Glyptodon + Doedicurus). A careful comparison of “U. antiquum” to well-characterized taxa with similar stratigraphic and geographic provenance reveals that no significant skull differences are observed from Plohophorus figuratus Ameghino. It seems likely that the alleged ornamentation pattern that characterizes this species was produced by a taphonomic alteration of the exposed surface of the osteoderms, a process broadly distributed in glyptodonts having a “rosette” ornamentation pattern. Finally, some characters present in the osteoderms of the type specimen of “U. antiquum” suggest that it may be a juvenile specimen. In summary, “U. antiquum” should be considered a junior synonym of P. figuratus.     

The Use of Albino Adult Hair and Blond Prepubertal Hair Yields Equivalent Results in an In Vitro Hair Perforation Test to Differentiate Between Different Dermatophytic Fungi

An in vitro hair perforation test is used to differentiate isolates of Trichophyton mentagrophytes and Trichophyton rubrum complexes because morphological criteria are insufficient. Here, we performed in vitro hair perforation tests using blond prepubertal hair and albino adult hair to determine whether they differentiate between fungal species. We tested 43 well-characterized dermatophyte strains, Arthroderma spp. [n = 4], Epidermophyton floccosum [n = 1], Microsporum spp. [n = 8], and Trichophyton spp. [n = 30], and examined hair perforation at 3–30 days postinoculation (p.i.). The perforation times were not significantly different between the two hair types (P > 0.05). The T. mentagrophytes complex strains perforated hair 4–5 days p.i., whereas T. rubrum complex strains perforated hair 13–30 days p.i., except for Trichophyton violaceum, which perforated hair after 6–7 days. Thus, the hair perforation test is highly sensitive (100 %) and specific (100 %) for differentiating T. mentagrophytes from T. rubrum complexes 5 days p.i. At 14 and 30 days, the sensitivity and negative predictive value of the test remained unchanged (100 %), but the specificity was reduced (64.3 and 14.3 %, respectively). Consistent with previous reports, we observed “perforating organs” of zoophilic Microsporum canis and geophilic Microsporum gypseum at 4 and 3 days, respectively. This paper offers a “low-cost” and “low-tech” alternative to differentiating dermatophyte species where standard morphological techniques fail and/or where molecular techniques are not a viable option.     

Comparison of the internal transcribed spacer region (ITS) of the ribosomal RNA genes in wild and cultivated two and six-rowed barleys (Hordeum vulgare L.)

The ITS region of the ribosomal RNA genes from two and six-rowed cultivated barley (Hordeum vulgare subsp. distichon and H. v. subsp. hexastichon, respectively), and its two and six-rowed wild relatives (H. v. subsp. spontaneum and H. v. subsp. agriocrithon, respectively) was isolated and sequenced. The entire ITS region is 598 bp in the two-rowed taxa (H. v. subsp. distichon and H. v. subsp. spontaneum) and 599 bp in the six-rowed ones (agriochriton and hexastichon). The ITS1 is 217 bp in the six-rowed barleys (H. v. subsp. agriochriton and H. v. subsp. hexastichon) and 218 bp in the two-rowed barleys (H. v. subsp. distichon and H. v. subsp. spontaneum). The 5.8S region is 163 bp in all studied H. vulgare taxa. The ITS2 region is 217 bp in the two-rowed barleys (H. v. subsp. distichon and H. v. subsp. spontaneum) and 219 bp in the six rowed ones (H. v. subsp. hexastichon and H. v. subsp. agriochriton). The ITS sequence data of the studied taxa and that of three other wild Hordeum species (H. murinum, H. marinum and H. chilense) were aligned and a phylogeny tree was reconstructed using the Lasergene Program. H. v. subsp. spontaneum was appeared as the ancestor of all other H. vulgare taxa.     

Taxonomic review of the genus Hypoatherina Schultz 1948 (Atheriniformes: Atherinidae)

The marine atherinid fishes of the genus Hypoatherina Schultz 1948 (Atherinidae: Atherinomorinae) were redefined from both morphological and molecular analyses, and eight of the ten included species were redescribed. In the molecular phylogeny, four regions of mitochondrial DNA were analyzed. The results of both trees of maximum likelihood and Bayesian analyses indicated the paraphyly of the former Hypoatherina. “Atherina” valenciennei and “Atherina” woodwardi, both formerly belonging to Hypoatherina, show closer relationships with Atherinomorus duodecimalis and Atherinomorus aetholepis. “Hypoatherina” celebesensis is also apart from the clade including the majority of Hypoatherina species. In contrast, H. panatela, formerly regarded as a member of the genus Stenatherina Schultz 1948, is included in the present Hypoatherina clade. The present molecular phylogeny of the genus Hypoatherina can be supported by morphology. The genus Hypoatherina is redefined by the following combinations of characters: ascending process of premaxilla long and slender, its height more than 2.7 times the maximum width; both anterior and posterior lateral processes of premaxilla narrow and deep, the anterior process almost the same as or slightly deeper than the posterior process; premaxilla not tapering posteriorly; posterior upper margin of dentary with a prominent process; upper posterior limb of dentary with round or somewhat angular posteroventral corner; anterior preopercular ridge with a deep notch just above the corner; anus situated posterior to or slightly anterior to appressed pelvic-fin tip in adults; dorsoventral height of exposed area in the midlateral scale (third) row wide, almost equal to the maximum height of the scale, and almost the same height as scales just above or below the midlateral row. The redefined Hypoatherina includes the following ten species: H. barnesi—widely distributed in the Indo-Pacific; H. gobio (lectotype designated herein)—restricted to the Red Sea; H. golanii—restricted to the Gulf of Aqaba, inner Red Sea; H. klunzingeri—from Mozambique to eastern South Africa; H. lunata—distributed in Japan and Indonesia; H. panatela—from western and central Pacific Ocean; H. temminckii—widely distributed in the Indo-West Pacific (neotype designated herein); H. tropicalis—restricted to the northeastern coast of Australia; H. tsurugae—occurring in Japan and South Korea; and H. uisila—distributed in the western and central Pacific Ocean.     

A new large species of gecko of the genus Hemidactylus Oken, 1817 (Reptilia: Sauria: Gekkonidae) from the Eastern Ghats,India

A new large species of gecko superficially resembling Hemidactylus maculatus Duméril & Bibron and its allied large Hemidactylus is described from Eastern Ghats. The new species, Hemidactylus kangerensis sp. nov. can be easily distinguished from members of the H. maculatus complex based on the number of femoral pores, i.e. 18–21 on each side, separated medially by 4 non-pored scales. A phylogenetic analysis based on partial sequence of mitochondrial gene cytochrome b shows that the gecko is a member of the “H. prashadi” clade and is sister to H. maculatus, from which it differs in an uncorrected pairwise sequence divergence of 10%. The discovery of a new large gecko from Eastern Ghats advocates the need for biodiversity assessment across the neglected Eastern Ghats.     

TLC screening of thraustochytrid strains for squalene production

Screenings of thraustochytrids (Labyrinthulomycetes) have been conducted for 176 strains isolated from various sites in the Asian region to investigate what type of species and strains accumulate high levels of squalene. Thin layer chromatography (TLC) screening for squalene production revealed that 38 strains were rated as “+” (high), 29 as “±” (medium), and 109 as “?” (low). Further, high performance liquid chromatography analysis strongly supported the TLC screening results. Besides the 18W-13a strain of Aurantiochytrium sp., which was previously recognized as a squalene-rich strain, several strains produced squalene at approximately 1 g L^?1 of culture volume. Squalene production was strongly related to locality, colony color, and phylogenetic clade. Most strains with “+” squalene spots were isolated from Okinawa, a subtropical region of Japan, while the strains with “±” and “?” squalene spots were isolated from wide geographical regions from tropical to subarctic. Approximately half the strains with orange colonies on GTY medium plates produced a high amount of squalene, whereas the other strains with different colors showed less or no squalene spots on TLC. All the squalene-rich strains were assigned to the Aurantiochytrium clade. Overall, our results suggest that (1) the thraustochytrids show tendentious locality in terms of squalene production, (2) a relationship exists between the metabolic synthesis of carotenoid pigments and squalene production, and (3) the Aurantiochytrium clade may have evolved to accumulate squalene.     

Late Miocene Proboscidea (Mammalia) from Macedonia and Samos Island, Greece: preliminary results

The abundant Late Miocene proboscidean remains of Greece have never been studied in detail and compared with those of Eurasia in order to determine their taxonomy and their biostratigraphical and palaeoecological significance. The first results of such study are given in this article. During the past decades, several new proboscidean specimens have been added to the old collections, significantly enriching the available material. The Axios Valley (Macedonia, Greece) proboscidean fossils belong mainly to two species of Choerolophodon: C. anatolicus of early Vallesian age and C. pentelici of late Vallesian–Turolian age. Deinotherium giganteum is rare and recognized only in the late Vallesian locality Ravin de la Pluie of Axios Valley. A zygodont form has also been identified in the Turolian of Axios Valley, attributed to “Mammut” sp. The Late Miocene localities of Nikiti (Macedonia, Greece) revealed several remains of C. pentelici, which are similar to the Turolian ones of Axios Valley. The Samos proboscidean collection includes C. pentelici, “Tetralophodon” atticus, “Mammut” sp. and Deinotherium gigantissimum. The taxonomy of the Late Miocene peri-Mediterranean Choerolophodon is given, and the biostratigraphy and palaeoecology of the Greek Late Miocene proboscideans are discussed.     

Establishment and optimization of a regionally applicable maize gene-flow model

Because of the rapid development of transgenic maize, the potential effect of transgene flow on seed purity has become a major concern in public and scientific communities. Setting a proper isolation distance in field experiments and seed production is a possible solution to meet seed-quality standards and ensure adventitious contamination of products is below a specific threshold. By using a Gaussian plume model as basis and data recorded by meteorological stations as input, we have established a simple regionally applicable maize gene-flow model for prediction of the maximum threshold distances (MTD) at which gene-flow frequency is equal to or lower than a threshold value of 1 or 0.1 % (MTD_1%, MTD_0.1%). After optimization of the model variables, simulated outcrossing rate was a good fit to data obtained from field experiments (y = 1.156x, R ² = 0.8913, n = 30, P < P _0.01). In the process of model calibration, it was found that only 15.82 % of the total amount of the pollen released by each plant participated in the dispersal process. The variable “a” for genetic pollen competitiveness between donor and recipient was introduced into our model, for the “Zinuo18” and “Su608” used, “a” was 17.47. Finally, the model was successfully used in the spring maize-growing region of Northeast China. The range of MTD_1% and MTD_0.1% in this region varied from 10 m to 49 m and from 17 m to 125 m, respectively.     

Cytogenetic study on some Fritillaria species of Iran

Five species (14 ecotypes) belonging to three subgenera of ornamental-medicinal Iranian Fritillaria plant were chromosomally and karyotypically assessed, using squash technique and 1 % (w/v) aceto-orcein stain. All species were diploid (2n = 2x = 24) having mean chromosome length of 16.8 μm (14.2–18.6 μm). The satellites varied in number (1–4 pairs) and in size (1.27–3.01 μm): mostly locating on long arms. Four chromosome types (“m”, “sm”, “st”, “T”) formed 11 different karyotypic formulas: the latter is being reported for the first time in some ecotypes in either S1 or S4. Nine chromosomal parameters were calculated. ANOVA verified intra- and inter-specific chromosomal variation in examined Iranian Fritillaria species. Twelve different methods were used to assess the degree of karyotype asymmetry. Among those, one qualitative (Stebbins classification) and seven quantitative (TF %, CV_TL, DI, AsK %, A2, Rec, CG %) parameters verified that S2 and S5 species were recognized as having the most asymmetrical and symmetrical karyotypes, respectively.     

Reproductive biology of an alpine orchid Phaius delavayi

The genus Phaius is distributed mainly in pantropical areas, and most species reported are autogamous. In contrast, Phaius delavayi, an alpine plant endemic to temperate zones of China, is strictly pollinator dependent. Only female workers and males of the bumblebee species B. hypnorum and B. lepidus are effective pollinators. The pollinaria are deposited on top of the head or the prothorax of the pollinators. The different depositions of pollinaria are regarded as an adaptation of the chamber dimensions of the flower to the size of the pollinator. The natural fruit set reported here (24.6–37.1%) suggests that pollination is successful in the deceptive P. delavayi. The yellow perianth and sweet floral scent of P. delavayi are important cues in attracting pollinators, while flowering time and mass population are associated with the process of “learning and avoidance” of the rewardless flowers. This orchid, therefore, employs a deceptive pollination strategy with diverse pollinators and pollinaria depositions rather than autogamy as do its congeners.     

The low fertility of Chinese white poplar: dynamic changes in anatomical structure, endogenous hormone concentrations, and key gene expression in the reproduction of a naturally occurring hybrid

Key message

We report that low fertility during intraspecific hybridization in Chinese white poplar was caused by prefertilization barriers, reduced ovules, and embryonic abortion. Hormone concentrations and gene expression patterns were also evaluated during the fertilization process.

Abstract

Hybrid vigor holds tremendous potential for yield increases and trait improvement; however, some hybridization combinations within Populus show very low fertility. To explore the causes of this low fertility in intraspecific hybridization of Chinese white poplar, we examined anatomical structure, hormone levels and expression of key genes in two unique crossing combinations of Populus × tomentosa “Pt02” × P. × tomentosa “LM50”, and (P. × tomentosa × P. alba cv. bolleana “Ptb”) × P. × tomentosa “LM50”. The seed set potential in the intraspecific hybridization P. × tomentosa “Pt02” × P. × tomentosa “LM50” was quite low, which was likely caused by prefertilization barriers, reduced ovule numbers, and embryonic abortion in ovaries. During intraspecific hybridization, we found reduced indoleacetic acid (IAA) in pistils, which may cause pollen tube deformations and increased IAA in heart-stage embryos, which may affect embryo development. Gibberellin A3 (GA3) decreased from the zygote dormancy stage to globular-stage embryos, which may be caused by failure of fertilization in specific embryos. The maximum zeatin (Z) concentration was found in heart-stage embryos, but Z concentrations quickly decreased, which may affect endosperm development. Increasing concentrations of abscisic acid (ABA) during zygote dormancy and eight-cell proembryo stages likely induced abscission of the infructescence. High ABA concentrations also regulated embryo maturity. Measurement of genes expression showed that high expression of SRK and/or SLG may result in rejection of pollen by stigmatic papillae through a mechanism, reminiscent of self-incompatibility. Also, low expression of LEC1 and FUS3 may cause embryonic abortion. Identification and eventual bypassing of these barriers may allow future genetic improvement of this key woody crop species.     

Electrophysiological and behavioural responses of the housefly to “sweet” volatiles of the flowers of Caralluma europaea (Guss.) N.E. Br.

In sapromyiophilous plants, up to date, long range attraction of fly pollinators has been thoroughly investigated and attributed to “fetid” floral compounds, while the “sweet” floral scent fraction has not been specifically investigated and its role has received little attention. The aim of the present study was to verify if terpenoids, which are the main compounds of the floral bouquet of Caralluma europaea, play a role in the attraction of its pollinator Musca domestica. Terpinolene, α-terpinene and linalool, described as the three main volatiles of the flowers of C. europaea, were evaluated in electrophysiological investigations and blends of these compounds as well as the whole fresh flowers were used in behavioural assays. Antennae of housefly adults showed positive dose-dependent responses to all the chemicals tested. Houseflies were attracted by the odour of the fresh flowers and by the reconstructed terpenoid blend at the dose of 100 μg. At the dose of 10 μg, the blend did not produce any attraction. The results of the present study support the hypothesis that terpinolene, α-terpinene and linalool emitted by C. europaea flowers are involved in pollinator attraction and demonstrate the importance of the “sweet” scent in this sapromyiophilous species.     

Diversity within the <Emphasis Type="Italic">Hygrophorus agathosmus</Emphasis> group (Basidiomycota,Agaricales) in Northern Europe

The North European species of the Hygrophorus agathosmus group in subsection Tephroleuci were studied. Three new species are identified based on morphology, ecology and sequence data. Two species are associated with Pinus spp. One of these is described here as H. suaveolens, while the other one is only known from one locality in the Nordic countries and seems to have a more South European distribution range. A closely related sister species to H. agathosmus is identified based on ITS sequence data, H. cf. agathosmus. It is confirmed to have an intercontinental distribution range and to be associated with Picea spp. probably on more acidic to neutral soil, whereas H. agathosmus s.s. has a more limited North-East European distribution range and occurs in older and rich Picea abies forests. A neotype for H. agathosmus is here selected from South Sweden. Hygrophorus agathosmus f. albus and H. agathosmus f. aureofloccosus are confirmed as forms. No genetic differences in the ITS region between specimens with grey cap colour and the two forms were observed. A key to the species in Northern Europe is provided.