Single step molecular characterization of morphologically similar black truffle species

Species-specific internal ITS primers that amplify polymerase chain reaction (PCR) products of different lengths were selected to distinguish the morphologically similar ectomycorrhizal fungi T. melanosporum, T. brumale and T. indicum by aligning their internal transcribed spacer sequences and taking into account any incidence of intraspecific variability. In multiplex PCR experiments, the species-specific primers yielded the expected amplicons on template DNA isolated from the above mentioned species, while there was no amplification in PCR reactions carried out on fungal DNA from competing truffle species and host plants.     

Allozyme comparison of four littorinid species morphologically similar to Littorina sitkana

Eight species of the genus Littorina were hitherto recognised in the north-western region of the Pacific Ocean: L. sitkana, L. brevicula, L. mandshurica, L. squalida, L. aleutica, L. naticoides, L. kasatka and L. subrotundata. Using allozyme electrophoresis it has been demonstrated that, in the Kurile Islands, three of these species (L. sitkana, L. subrotundata and L. kasatka) co-occur, together with a fourth, still undescribed species (L. sp.). These four species were compared at 16 loci coding for 13 enzymes. All species were easily distinguished by diagnostic enzyme markers. The mean genetic distances and ranges between species pairs are: L. sitkana and L. sp. D = 0.622 (0.561–0.741), L. sitkana and L. subrotundata D=0.981 (0.821–1.110), L. subrotundata and L. sp. D=0.975 (0.955–0.995). The genetic distance between L. kasatka and each of the other three species was greater than 1 (range 1.123–2.087). These data suggest that L. sitkana, L. subrotundata and L. sp could be members of a species complex; according to current classifications these three belong to the subgenus Neritrema. However, the genetic distance between L. kasatka and L. sitkana is much greater than between L. sitkana and other Neritrema species, and thus supports the classification of L. kasatka in the subgenus Littorina.     

Taxonomy of Quercus crassifolia (Fagaceae) and morphologically similar species in Mexico

In this work, ten species morphologically similar to Q. crassifolia are circumscribed based on a phylogenetic species concept. Examination of these species suggests that Q. dysophylla and Q. radiata are of hybrid origin; the putative parents of Q. dysophylla are Q. crassifolia and Q. crassipes, while Q. radiata is likely the result of hybridization between Q. coccolobifolia and Q. conzattii or Q. urbanii. A lectotype of Q. crassifolia is designated here. Dichotomous keys, as well as distribution ranges and photographs, are presented.     

Taxonomy of Pluteus eugraptus and morphologically similar taxa

The status of the taxa morphologically similar to Pluteus eugraptus (Basidiomycota, Agaricales) was investigated with morphological and molecular (ITS region) characters. This group of species belongs in Pluteus sect. Celluloderma based on morphological and molecular characters. Two species, Pluteus multiformis, from Spain and Pluteus eludens from Madeira, Portugal, Russia and USA, are described as new. Both species share pigmented cheilocystidia and a pileipellis composed of both clavate-spheropedunculate and elongated elements with P. eugraptus, but they can be separated based on the characteristics of the cystidia and pileipellis. Pluteus multiformis is characterized by the scarce pleurocystidia, clavate cheilocystidia and caulocystidia and highly polymorphic elements of the pileipellis. Pluteus eludens is characterized mainly by utriform pleurocystidia. Pluteus eugraptus is known only with certainty from the type collection (Sri Lanka), which has been re-examined here, and it is characterized by narrowly lageniform pleurocystidia. Phylogenetic analyses based on ITS region sequence data supported the separation of P. multiformis, P. eludens and an additional collection from Japan that likely represents the true P. eugraptus.     

Isozyme and allozyme markers distinguishing two morphologically similar, medically important Mastomys species (Rodentia: Muridae)

Background  

Two common southern African mice species, Mastomys coucha and M. natalensis, are widely distributed throughout the subregion and overlap in many areas. They also share a high degree of morphological similarity, making them impossible to distinguish in the field at present. These multimammate mice are documented carriers of serious disease vectors causing Lassa fever, plague and encephalomyocarditis, which coupled to their cohabitation with humans in many areas, could pose a significant health risk. A preliminary study reported the presence of isozyme markers at three loci (GPI-2, PT-2, -3) in one population each of M. coucha and M. natalensis. Two additional populations (from the Vaal Dam and Richards Bay) were sampled to determine the reliability of these markers, and to seek additional genetic markers.     

Biochemical genetic markers to identify two morphologically similar South African Mastomys species (Rodentia: Muridae)

The two common southern African mice species (Mastomys coucha and M. natalensis) are morphologically almost identical, making field identification impossible at present. Specimens from two localities were collected and tissue and blood samples taken. The habitat type of each locality was studied, and a distribution map compiled. A definite correlation between biome-type and species range was found to be present. Three isozyme markers were identified: glucose phosphate isomerase in liver, and two general (non-specific) protein coding loci in muscle. In addition, we also identified species characteristic haemoglobin components in both species. This is the first study to report genetic variation within, and differentiation between these species. Our results are of medical importance because Mastomys coucha carries bubonic plague and M. natalensis carries Lassa Fever.     

Sharing the same space: foraging behaviour of saproxylic beetles in relation to dietary components of morphologically similar larvae

1. Saproxylic beetles (Coleoptera) have particular requirements for dead wood. Little is known about their larval development, especially within intermediate and late stages of dead wood decay. 2. We studied the natural foraging behaviour of three saproxylic beetles with morphologically similar (i.e. convergent) larvae and similar habitat requirements –Cucujus cinnaberinus Scopoli (Cucujidae), Pyrochroa coccinea L., and Schizotus pectinicornis L. (Pyrochroidae) – examining dietary components using visual gut content analysis. 3. There were few significant differences in diet among the study species, but there were significant differences for the different dietary components in individual species. Guts of all three species included more plant and fungal material than animal. We also observed that with increasing body size larvae foraged significantly less on fungal material. We observed significant differences among species and selection with respect to body size during the overwintering period. Larvae swallowed more animal and less fungal material with increasing body size. 4. The study species seem to be opportunistic foragers, varying their foraging capability according to seasonality and age. They probably generally foraged woody material infested by filamentous fungi (i.e. xylomycetophagy), swallowing other material that is in their way while foraging, including animal parts and small arthropods. 5. We found that morphologically similar larvae do not differ much in their foraging behaviour even when sharing the same habitat. Larval size seems to be a useful predictor for foraging behaviour of saproxylic beetles within the same guild. The study species are probably occasional predators with less ability to forage on other animals than previously suggested.     

Phylogeography and species discrimination in the Paracalliope fluviatilis species complex (Crustacea: Amphipoda): can morphologically similar heterospecifics identify compatible mates?

We examined the phylogeographic patterns of Paracalliope fluviatilis (Amphipoda) over its entire range and the prevalence of mate discrimination in laboratory mate choice tests using genetically distinct populations. We predicted that mate discrimination would increase as the level of genetic divergence increased between populations. Thirty different haplotypes were found with pairwise sequence divergences in the range 1–23.5% between locations. Individuals were selected from seven genetically distinct populations and males were presented with ‘local’ (same population) or ‘foreign’ (genetically divergent) females. Males were more likely to pair with local than foreign females but there was no evidence of a gradual increase in discrimination. Discrimination became most prominent (approximately 5 : 1 local : foreign) when genetic divergences exceeded approximately 20%; matings between divergent individuals also resulted in significantly fewer females producing eggs. We suggest that: (1) this abrupt shift in discrimination occurs because individuals from different, but similarly divergent, clades rely on different recognition cues (e.g. moulting pheromones with disparate chemical signatures) that trigger recognition and subsequent discrimination of incompatible mates; (2) geological history associated with sea level changes and a series of isolation events may be responsible for the patterns of discrimination that we observed; and (3) amphipods may be more genetically variable relative to other invertebrate taxa. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 196–205.     

Molecular and morphologically distinct Scytosiphon species (Scytosiphonales,Phaeophyceae) display similar antioxidant capacities1

The taxonomic status of an alga with complanate thalli, occurring in central Chile and belonging to the genus Scytosiphon, was elucidated. Morphological and molecular features demonstrated that, in addition to the known and widespread constricted S. lomentaria (Lyngb.) Link that occurs along the Chilean coast, there is a Scytosiphon with complanate thalli that occurs only in central Chile—S. gracilis Kogame. Morphological analyses of this previously unreported complanate Scytosiphon showed thalli without constrictions, coherent plurilocular sporangia without ascocysts, and phaeophycean hairs arising from cortical cells. Furthermore, sequences of the internal transcribed spacer (ITS1 and ITS2) regions of the complanate Scytosiphon were 99.8% and 100% identical to those detected in S. gracilis from Korea. ITS‐based comparative analyses showed that complanate Scytosiphon grouped in a different clade than S. lomentaria and S. tenellus Kogame from various parts of the world, including Chilean species. Moreover, molecular analyses suggest the occurrence of two distinct ITS types of S. lomentaria in northern Chile, corresponding to the Korean and Greek types. On the other hand, biochemical analyses of copper‐induced antioxidant responses in S. gracilis and S. lomentaria showed an identical increase in antioxidant enzyme activities. These results suggest that copper tolerance might be a constitutive trait in these species of Scytosiphon.     

Chromosomal and protein evolution in morphologically similar species of Praomys sensu lato (Rodentia, Muridae)

Evidence of extensive chromosomal evolution in a biologically and economically important group of African murids of the Praomys/Mastomys complex was provided by examination of G- and C-band chromosomal data on P. coucha (2n = 32), P. fumatus (2n = 38), P. hildebrandti (2n = 32), P. jacksoni (2n = 28), P. misonnei (2n = 36), and P. cf. tullbergi (2n = 35). A coding system was developed for the chromosomal characters, and analyses were performed by a computer program to find the shortest tree with a minimum of 35 autosomal rearrangements (pericentric inversions, complex translocations, centric fusions, centric fissions, tandem fusions, euchromatic additions, and heterochromatic additions). The resulting phylogenetic hypothesis differs from traditionally accepted hypotheses regarding this complex group of rodents. The cytogenetic data show that 1) there is no support for the dichotomy of Mastomys/Praomys previously based on morphology, 2) the 2n = 32 species from eastern Africa (P. hildebrandti) is distinct from the 2n = 32 species from southern Africa (P. natalensis), and 3) there is a close association between P. jacksoni and P. cf. tullbergi. Polyacrylamide gel electrophoresis of liver membrane proteins demonstrated few differences in protein mobilities between species and even fewer between individuals of the same species taken from different habitats and localities in Kenya. Monoclonal antibodies produced against liver proteins of one species and tested for reactivity to other species confirmed the evolutionary similarity of species of this complex. This immunologic approach may provide a robust data set for future phylogenetic studies of muroid rodents.(ABSTRACT TRUNCATED AT 250 WORDS)     

Geographic variation in hybridization and ecological differentiation between three syntopic,morphologically similar species of montane lizards

To understand factors shaping species boundaries in closely related taxa, a powerful approach is to compare levels of genetic admixture at multiple points of contact and determine how this relates to intrinsic and extrinsic factors, such as genetic, morphological and ecological differentiation. In the Australian Alps, the threatened alpine bog skink Pseudemoia cryodroma co‐occurs with two morphologically and ecologically similar congeners, P. entrecasteauxii and P. pagenstecheri, and all three species are suspected to hybridize. We predicted that the frequency of hybridization should be negatively correlated with genetic divergence, morphological differentiation and microhabitat separation. We tested this hypothesis using a mitochondrial locus, 13 microsatellite loci, morphological and microhabitat data and compared results across three geographically isolated sites. Despite strong genetic structure between species, we detected hybridization between all species pairs, including evidence of backcrossed individuals at the two sites where all three species are syntopic. Hybridization frequencies were not consistently associated with genetic, morphological or ecological differentiation. Furthermore, P. entrecasteauxii and P. pagenstecheri only hybridized at the two sites where they are syntopic with P. cryodroma, but not at the largest site where P. cryodroma was not recorded, suggesting that P. cryodroma may serve as a bridging species. This study reveals the complex dynamics within a three species hybrid zone and provides a baseline for assessing the impact of climate change and anthropogenic habitat modification on future hybridization frequencies.     

Phylogenetic relationships on 14 morphologically similar species of Pucciniastrum in Japan based on rDNA sequence data

We analyzed the phylogenetic relationships of 49 specimens comprising 14 morphologically similar species of Pucciniastrum distributed in Japan based on the sequence data of the large subunit rDNA (D1/D2), 5.8S rDNA, and internal transcribed spacer (ITS) regions. Neighbor-joining and parsimony analyses generated six major groups for both the D1/D2 and ITS regions. Pucciniastrum circaeae and P. epilobii formed a single group. P. hydrangeae-petiolaris, P. coryli, P. fagi, P. hikosanense, P. tiliae, and P. boehmeriae were each a distinct clade, and P. fagi formed a close relationship with P. hikosanense. However, these analyses did not support the monophyly of the following species: P. kusanoi, P. actinidiae, P. corni, P. styracinum, P. yoshinagai, and P. miyabeanum. Contribution no. 201, Laboratory of Plant Parasitic Mycology, Graduate School of Life and Environmental Sciences, University of Tsukuba, Japan     

Host preferences and behaviour of oxpeckers: co-existence of similar species in a fragmented landscape

The host preferences and behaviour of red-billed and yellow-billed oxpeckers (Buphagus erythrorynchus and Buphagus africanus) were studied at two locations in Kenya and combined with previously published data from East and South Africa. Red-billed oxpeckers are generally more common and have a greater niche breadth with respect to host preferences than yellow-billed oxpeckers, possibly due to the behavioural dominance of the latter. Otherwise, the host preferences are remarkably similar: both prefer larger species of ungulates and are found predominantly on species with manes. There are also no significant differences in the time–activity budgets of the two species controlling for site and host species, and differences in the proportion of time spent on different body parts of hosts are small in absolute magnitude even when statistically significant. Despite these and other basic ecological similarities, no evidence for competitive displacement was detected by comparing host preferences in areas of sympatry with areas of allopatry, even though each species appears to depress the population size of the other when sympatric. Traditional single-community competition theory cannot explain the geographical ecology of these two species. Instead, co-existence appears to be dependent on the patterns of extinction and colonization characterized by metapopulations distributed among a fragmented habitat.     

Molecular species identification of morphologically similar mussel larvae reveals unexpected discrepancy between relative abundance of adults and settlers

Understanding the relative importance of larval supply vs. post-settlement mortality underlies studies of marine invertebrate recruitment, yet is often hampered by researchers' inability to identify species among morphologically similar larvae or early juveniles. In New Zealand, two species of co-occurring intertidal mytilid mussels have morphologically indistinguishable settlers: the blue mussel Mytilus galloprovincialis, which is often numerically dominant in the mid-zone of the rocky intertidal, and the ribbed mussel Aulacomya atra maoriana which is often much less abundant. In this study, we obtained samples of newly settled mussels from 6 sample dates April-May 2005 from the rocky intertidal in Wellington Harbour, New Zealand. We used PCR-RFLP of the cytochrome c oxidase subunit I (COI) mitochondrial gene region to identify settlers to species. Of a total of 224 settlers that could be identified, 64% were identified as Mytilus galloprovincialis and 36% as Aulocomya atra maoriana. The percentage of A. atra maoriana in the samples was unexpectedly high and ranged from 22–50% among the sample dates. This study reinforces the need to quantify larval supply at the species level to understand the relative importance of pre- and post-settlement mortality, and also demonstrates the usefulness of the COI region as a species-specific marker for identifying mussel larvae and juveniles.     

Molecular systematics of Ceratostomella sensu lato and morphologically similar fungi

The systematic position and phylogenetic relationships of Ceratostomella sensu lato and phenotypically similar fungi using comparative morphological and culture studies and phylogenetic analyses of the nuclear large- and small-subunit ribosomal DNA were explored. In the light of inferred phylogenies and morphological data the genus Ceratostomella is redescribed, the generic concept is emended and four species are accepted (viz. C. cuspidata, C. pyrenaica, C. rhynchophora and C. rostrata). A new genus Xylomelasma is introduced and delimited from Ceratostomella, with two new species described (viz. X. novaezelandiae and X. sordida). In culture species of both Ceratostomella and Xylomelasma produced sterile mycelium. The genus Lentomitella with a phaeoisaria-like anamorph formed in vitro is reinstated to encompass taxa formerly attributed to the broadly perceived Ceratostomella with three accepted species (viz. L. cirrhosa, L. crinigera and L. tomentosa). Lentomitella and Ceratostomella are clearly distinguishable by the morphology of asci, ascospores and centrum. Lentomitella is compared to phenotypically similar Ceratosphaeria, which formed a harpophora-like anamorph in vitro. In the present phylogenies Ceratostomella, Ceratosphaeria, Lentomitella and Xylomelasma are shown as clearly separate genera belonging to three different groups of perithecial ascomycetes. Ceratostomella, Lentomitella and Xylomelasma reside within a large unsupported clade consisting of members the Ophiostomatales, the freshwater Annulatascaceae and a group of nonstromatic, terrestrial taxa. Ceratosphaeria is well supported within the Magnaporthaceae. The systematic value of morphological characters of ascospores, paraphyses, asci, centrum and conidiogenesis in segregating taxa from Ceratostomella sensu lato and their relatives is discussed.     

Feeding and defecation of Rhodnius (hemiptera: Reduviidae) fed human blood

Feeding and defecation behavior of Rhodnius prolixus Stal, 1859, R. robustus Larrousse, 1927, R. neivai Lent, 1953 and R. pictipes Stal, 1872, artificially fed on human blood, were studied under laboratory conditions. In all species, first instar nymphs did not defecate in the first 30 minutes after feeding. R. pictipes did not accept artificial feeding but fed directly on humans. Nymph and adult R. prolixus had a higher defecation index (DI) than other species; third instar nymphs had the highest DI = 1.62. In all instars, most individuals accepted the food in 3 Pounds minutes and finished feeding in less than 15 minutes.     

Ubiquinone systems of the genus Cladosporium and morphologically similar taxa

Abstract We measured adenosine deaminase (ADA) activity in a guinea pig model of Legionella pneumophila infection. Female Hartley guinea pigs were inoculated intraperitoneally with one-quarter of the LD₅₀ dose of L. pneumophila Philadelphia-1 strain. Control groups were inoculated with clinical isolates of Staphylococcus aureus, Streptococcus pneumoniae, Haemophilus influenzae or Klebsiella pneumoniae . Each group consisted of 5 animals. ADA activity in plasma was assayed calorimetrically before and at various intervals after infection by measuring the amount of ammonia produced after adnosine was added to plasma samples. ADA activity before inoculation was 25.6±6.0 IU/1, it reached 174.4±60.0 IU/1 on day 3 after inoculation of L. pneumophila . ADA activity returned to normal levels on day 14. ADA activity did not increase significantly in guinea pigs infected with the other types of bacteria. These findings suggest that measurement of plasma ADA activity may be useful for the diagnosis of Legionella infection.     

Deep genetic differentiation between two morphologically similar species of wolf herrings (Teleostei,Clupeoidei, Chirocentridae)

Wolf herrings (Chirocentridae; Clupeoidei) are commonly found in local fish markets throughout the Indo‐West Pacific region where they constitute an auxiliary source of food and income for local communities. The validity of the two species of wolf herrings, Chirocentrus dorab Forsskål, 1775 and C. nudus Swainon, 1839, is only supported by slight morphological differences. The identification of either species is challenging, especially for juveniles, and precludes accurate assessments of these natural resources at a species level. As a step towards gaining better knowledge of the genetic structure of these fishes, we examined genetic differentiation between these two species by reconstructing their entire mitogenomic sequences using high‐throughput sequencing technology. We found that the mitogenome of each species shared the same gene content and order that were the same for those found in most other teleost fishes. Despite their high morphological similarity, these two species of Chirocentrus were genetically well differentiated (p‐distance = 16.3% at their cytochrome oxidase I). A mitogenomic time‐calibrated phylogenetic analysis showed that wolf herrings originated about 35 million years ago, and they represent a case of morphological stasis. Furthermore, comparison of published and newly determined mitochondrial COI barcode region sequences from 22 individuals revealed species‐level cryptic genetic diversity within C. dorab. Altogether, these mitochondrial data are effective in discriminating species within this genus and informing population genetic relationships within species of wolf herrings.