Morphological data and the first molecular data are provided for four species of the trematode family Bucephalidae Poche, 1907 from marine and freshwater teleost fish species of East Asia. A new species, Rhipidocotyle husi n. sp., was isolated from Huso dauricus from the Amur River, Russia. Adult worms of this species were distinguished from their congeners Rhipidocotyle illense and Rhipidocotyle kovalai by morphological analysis. Three other known species were identified: Bucephalus skrjabini and Prosorhynchus cf. squamatus were detected in Siniperca chuatsi from the Amur River and in Myoxocephalus spp. from the Okhotsk Sea, Russia, respectively, while Prosorynchoides karvei was extracted from Strongylura strongylura from Halong Bay, Vietnam. The 28S ribosomal DNA (rDNA)-based phylogenetic analysis showed that the new species formed a shared polytomy clade with Rhipidocotyle fennica. Phylogenetic analysis of all available molecular data showed that four genera, namely Rhipidocotyle, Bucephalus, Prosorynchoides and Prosorhynchus, are para- or polyphyletic. Molecular-based phylogenetic analysis of morphologically validated bucephalid species indicated that three genera – Rhipidocotyle, Bucephalus and Prosorynchoides – were monophyletic. The genus Prosorhynchus maintained paraphyly, and P. cf. squamatus was more closely related to Dollfustrema spp. than to other Prosorhynchus spp. These findings do not exclude the possibility that representatives of Dollfustrema and P. cf. squamatus belong to the same genus.

Bopyrid isopods of the genus Probopyrus are well-known parasites of freshwater prawns of the genus Macrobrachium. The parasitism of coastal populations of Macrobrachium amazonicum by Probopyrus bithynis, for example, has been documented since the late 1980s. Despite this, molecular data on different populations are not available for any Probopyrus species. The present study is the first to describe Probopyrus populations from distinct regions of the Amazon basin based on sequences of two genes, the mitochondrial cytochrome oxidase C subunit I (COI) and the nuclear 18S ribosomal DNA (18S rDNA) gene. The analyses indicated the presence of two Probopyrus species, each parasitizing either the coastal or the inland populations of M. amazonicum. The results indicated the potential use of the COI barcode for the identification of Probopyrus species. We discuss the potential implications of the findings for the taxonomy of Probopyrus bithynis and other species of the genus Probopyrus.

Echinococcosis is a zoonotic parasitic illness that can cause significant disabilities, and even death for sick people. The disease is caused by the larval stage of cestodes belonging to the Echinococcus genus. In this study, multiple hydatid cysts were excised from an infected porcine liver. The identification of the parasitic species was made by the morphometric assessment of rostellar hooks and molecular detection of ribosomal DNA extant in protoscoleces of the hydatid sand. Rostellar hooks presented an average length of 27.4 µm by optical microscopy. Parasite DNA were detected in samples of hydatid sediment and positive controls by polymerase chain reaction. In conclusion, Echinococcus granulosus was recognized in samples of porcine hydatid cysts by microscopic observation, and the E. granulosus sensu lato strain E. canadensis G6/G7 was identified by molecular assay.

Asynchronous flowering was noted in a recently discovered infestation of siam weed in north Queensland. This may indicate some genetic diversity in the infestation, increasing concerns about the origin of the infestation. Internal transcribed spacer 1 (ITS1) sequence data were obtained for siam weed individuals from north Queensland, Indonesia, Thailand, South Africa, Ivory Coast, Brazil, Colombia, and the U.S. The ITS1 region is 258 base pairs long, and the populations that flower at different times in north Queensland differ by four base substitutions. The genotype common in north Queensland is also reported throughout the native and introduced ranges. The other genotype is reported only in north Queensland and southern Brazil. These data, in conjunction with prior investigations into possible origins, indicate that Brazil is the most likely source of the infestation in Australia.

A triazine-resistant (TR) biotype of Powell amaranth was discovered in 1989 in a potato field treated with metribuzin. A survey of all agricultural counties in Idaho showed that the TR Powell amaranth infestation was localized in the southeastern corner of Gooding county in southern Idaho. To determine the mechanism of triazine resistance, I50 values for inhibition of photosystem II were determined for atrazine, metribuzin, and diuron using thylakoids isolated from TR and triazine-susceptible (TS) biotypes. TR/TS ratios based on I50 values were 134 for atrazine, 62 for metribuzin, and 1.9 for diuron. Results of binding studies with atrazine and metribuzin were consistent with the I50 studies, indicating that resistance was due to reduced binding of triazines to the thylakoid membrane D1 protein. Sequencing the chloroplast psbA gene from TR and TS biotypes revealed a serine 264 to glycine change in the TR biotype. The mutation presumably resulted in reduced hydrogen bonding between triazine herbicides and the Dl protein.

Common ragweed and common cocklebur plants were collected at two sites each in Illinois, Minnesota, and Ohio to analyze intraspecific variability of the gene encoding acetolactate synthase (ALS). A 385-nucleotide fragment within the coding sequence of ALS was compared among 24 plants of each of these two species from the six locations. Common ragweed ALS was highly variable, with polymorphisms observed at 48 (12.5%) of the 385 nucleotides among the 24 plants. Despite the numerous nucleotide polymorphisms, only two inferred amino acid polymorphisms were identified. No apparent population structure was suggested by the ALS sequence data, indicating widespread gene flow consistent with the wind-pollinated nature of common ragweed. In contrast to common ragweed, no ALS polymorphisms were identified among the common cocklebur plants used in this study. As a basis for comparing the extremes observed between common ragweed and common cocklebur, ALS intraspecific variability also was investigated in 10 plants each of tall waterhemp and smooth pigweed. Normalized to the number of plants analyzed, the number of nucleotide polymorphisms for both tall waterhemp and smooth pigweed was greater than that in common cocklebur but less than that observed in common ragweed. Information on variability of herbicide target-site genes may be useful in predicting the likelihood for herbicide-resistance development. However, all four of the species investigated in this study have evolved resistance to ALS-inhibiting herbicides, despite the different levels of ALS variability observed.

Genetic markers have been used in horticulture and conservation biology to identify breeding lines, assess genetic diversity, and examine gene flow. In weed science, analysis of genetic markers is not as common, as the focus often lies in the development of control methods. This is unfortunate, because advances in genetic marker techniques may lead to innovative methods in controlling weedy plants. Microsatellites, random amplified polymorphic DNA (RAPDs), intersimple sequence repeats (ISSRs), amplified fragment length polymorphisms (AFLPs), DNA sequences, single nucleotide polymorphisms (SNPs), and derived cleaved amplified polymorphic sequences (dCAPS) have been used in studying genetics of weedy plants. Beyond assessing genetic diversity of weeds, markers have been used to examine gene flow, patterns of dispersal, ploidy levels, and relationships of weedy and nonweedy species, as well as identifying founder populations. Identification of closely related species may indicate the potential for hybridization, cross reactivity of chemical applications, or nontarget biological control effects. Furthermore, markers have been used in identification of mutations conferring herbicide resistance and identification of regions correlating fitness of weedy and hybrid plants in new habitats. Knowledge from genetic markers provides fundamental information that can be used in advancing chemical and biological control in weed management. This article serves as a review of these marker types and their application to weed science.

Recent advances in sequencing technologies (next-generation sequencing) offer dramatically increased sequencing throughput at a lower cost than traditional Sanger sequencing. This technology is changing genomics research by allowing large scale sequencing experiments in nonmodel systems. Waterhemp is an important weed in the midwestern United States with characteristics that makes it an interesting ecological model. However, very few genomic resources are available for this species. One half of a 70 by 75 picotiter plate of 454-pyrosequencing was performed on total DNA isolated from waterhemp, generating 158,015 reads of an average length of 271 bp, or a total of nearly 43 Mbp of sequence. Included in this sequence was a nearly complete sequence of the chloroplast genome, sequences of several important herbicide resistance genes, leads for simple sequence repeat (SSR) markers, and a sampling of the repeated elements (e.g., transposons) present in this species. Here we present the waterhemp genomic data gleaned from this sequencing experiment and illustrate the value of next-generation sequencing technology to weed science research.

Perennial pepperweed is an invasive plant species in North America, native to temperate Eurasia and northern Africa. Effective biological control depends upon correct taxonomic identification. Therefore, we investigated morphological and genetic data (cpDNA sequences and amplified fragment length polymorphisms [AFLP]) in its native range, where the species is at times treated as multiple taxa (L. latifolium, L. affine and L. obtusum). We also analyzed genetic data to determine the number and distribution of haplotypes and genotypes in the invaded range. Using Bayesian analysis, we found three clusters of AFLP genotypes in the native range, but little correlation between these clusters and morphological characters used to distinguish taxa. Also, we found combinations of morphological character states within many native range plants that are incompatible with current species descriptions, offering no support for splitting L. latifolium sensu lato into three species. In North America 97% of the genetic variation was among populations and there were only eight AFLP genotypes in 288 plants, suggesting few introductions or a severe bottleneck, and little or no creation of new genotypes since introduction. We found plants in the native range that are genetically similar (88 to 99%) to six of the eight invasive AFLP genotypes, suggesting that Kazakhstan and China are origins for much of the North American invasion.

Detection of repeated sequences within complete genomes is a powerful tool to help understanding genome dynamics and species evolutionary history. To distinguish significant repeats from those that can be obtained just by chance, statistical methods have to be developed. In this paper we show that the distribution of the number of long repeats in long sequences generated by stationary Markov chains can be approximated by a Poisson distribution with explicit parameter. Thanks to the Chen-Stein method we provide a bound for the approximation error; this bound converges to 0 as soon as the length n of the sequence tends to ∞ and the length t of the repeats satisfies n2ρt = O(1) for some 0 < ρ < 1. Using this Poisson approximation, p-values can then be easily calculated to determine if a given genome is significantly enriched in repeats of length t.

This paper describes a terebratelloid articulate brachiopod, Gyrothyris williamsi sp. nov., based on 95 specimens from seamounts on the Lord Howe Rise, Coral Sea, SW Pacific Ocean. The new species is attributed to Gyrothyris on the basis of (a) morphological and growth trajectory similarities; (b) phylogenetic analyses of an alignment of DNA sequence (∼2900-sites) obtained from nuclear-encoded small- and large-subunit ribosomal RNA genes (SSU and LSU); and (c) the presence of a distinctive, two-part deletion in the LSU gene. It is distinguished morphologically from Gyrothyris mawsoni and its subspecies by both internal and external morphology and by its isolated geographical distribution, which extends the patchy, known range of this genus to an area some 2000 km north of its previous northern limit around New Zealand. Phylogenetic analyses of the rDNAs and of mitochondrial cox1 gene sequences (663 sites) confirm previous indications that the New Zealand endemic terebratelloid genera form a clade (Neothyris (Calloria, Gyrothyris, Terebratella), but the position of Terebratella with respect to Calloria and Gyrothyris remains weakly established. These sequences disagree inexplicably about the closeness of the relationship between Neothyris parva and N. lenticularis. Analyses of the first sequences from Calloria variegata, a species restricted to the Hauraki Gulf, New Zealand, are consistent with the possibility that it originated locally, and recently, from C. inconspicua. Magellania venosa from S. America/Falklands joins with Antarctic Magellania fragilis and M. joubini to form an rDNA clade that excludes Terebratalia as the putative sister-group of the New Zealand terebratelloid clade. The cox1 (but not the rDNA) sequences of the New Zealand clade pass a test for clock-like rates of evolution, and maximum likelihood pairwise distances suggest that if genetic isolation between the ancestor of Antarctic Magellania and the last common ancestor of the New Zealand terebratelloid clade was initiated by separation of the Antarctic and New Zealand plates ∼90 Mya, isolation from M. venosa was initiated earlier, perhaps ∼145 Mya. However, in the simple phylogenetic reconstruction presented here from cox1 sequences, S. American and Antarctic Magellania spp. do not yield a well-supported clade, perhaps because of differences in base composition.

Eight South American geographical populations of the parasitoid Microctonus hyperodae Loan were collected in South America (Argentina, Brazil, Chile and Uruguay) and released in New Zealand for biological control of the weevil Listronotus bonariensis (Kuschel), a pest of pasture grasses and cereals. DNA sequencing (16S, COI, 28S, ITS1, β-tubulin), RAPD, AFLP, microsatellite, SSCP and RFLP analyses were used to seek markers for discriminating between the South American populations. All of the South American populations were more homogeneous than expected. However, variation in microsatellites and 16S gene sequences corroborated morphological, allozyme and other phenotypic evidence of trans-Andes variation between the populations. The Chilean populations were the most genetically variable, while the variation present on the eastern side of the Andes mountains was a subset of that observed in Chile.

The four species of Algyroides Bibron & Bory, 1833 form part of the relatively plesiomorphic Palaearctic clade of lacertids comprising Lacerta and its allies. An estimate of phylogeny based on DNA sequence from parts of the 12S and 16S rRNA mitochondrial genes confirms the monophyly of the genus already suggested by several morphological features. The molecular data also indicates that relationships within the clade are: (A. nigropunctatus (A. moreoticus (A. fitzingeri, A. marchi))); this agrees with an estimate of phylogeny based on morphology that assumes the taxon ancestral to Alygroides was relatively robust in body form, and not strongly adapted to using crevices. Initial morphological evolution within Algyroides appears to involve adaptation to crypsis in woodland habitats. The most plesiomorphic form (A. nigropunctatus) is likely to have originally climbed extensively on tree boles and branches and there may have been two subsequent independent shifts to increased use of litter and vegetation matrices with related anatomical changes (A. moreoticus, A. fitzingeri), and one to increased use of crevices (A. marchi). Some members of Algyroides are strikingly similar in superficial morphology to particular species of the equatorial African genus Adolfus. This resemblance results from a combination of many shared primitive features plus a few independently acquired derived ones that are likely to give performance advantage in the relatively similar structural niches that these forms occupy. This study provides evidence that: (1) the use of a combination of molecular and morphological data may sometimes allow the estimation of ancestral anatomical features when these are otherwise unknown; (2) process considerations may permit a choice to be made in cases of character evolution where tree topology means that equally parsimonious alternatives exist; such decisions about character evolution may allow ecological shifts to be similarly assessed; (3) parallel evolution in ecological analogues may involve relatively few characters.

In this paper the Leishmania (Viannia) braziliensis complex is defined as containing all species of the actual subgenus Viannia. Organisms of the L. (V) braziliensis complex are the causative agents of localized human cutaneous and mucocutaneous leishmaniasis in South America, much of Central America and some areas of North America. In our search for better species and subspecies diagnostic probes we focused our research on repetitive DNA, since it provides a greater number of target sites for hybridization. In this work we report the isolation and sequencing of a 1·8 kb DNA region, LbJ38, which is probably tandemly repeated or dispersed at least 4 times along one chromosome and is naturally present in L. (V) braziliensis genomic DNA. This region contains microsatellites and simple repeat DNA sequences and was isolated by screening a genomic DNA cosmid library with complex- and species-specific probes. No homology was found with other Leishmania microsatellite or repetitive DNA. The utility of this repetitive sequence and primers derived from it in the identification of L. (V) braziliensis is demonstrated. As far as we are aware, this is the first report of sequence characterized repetitive microsatellite and GC rich simple repeat DNA from the nuclear genome of New World Leishmania.

Two cDNAs encoding cysteine proteinases were isolated from a cDNA library constructed from feeding females of Heterodera glycines. The library was screened with a cysteine proteinase gene fragment originally amplified from cDNA of H. glycines. Database searches predict that 1 cDNA (hgcp-I) encodes a cathepsin L-like proteinase, while the second (hgcp-II) encodes a cathepsin S-like enzyme. Both predicted proteins contain a short secretion signal sequence, a long pro-peptide and a mature protein of 219 amino acids. Southern blot analysis suggests that the cathepsin S-like enzyme, HGCP-II, is encoded by a single-copy gene in contrast to the cathepsin L-like proteinase, HGCP-I which may have 2 homologues. The regions encoding the mature proteinases were cloned into an expression vector and recombinant protein produced in E. coli. HGCP-I was shown, after refolding, to cleave the synthetic peptide Z-Phe-Arg-AMC, and this activity could be inhibited by the engineered rice cystatin Oc-IΔD86. HGCP-II showed no activity against the synthetic substrates tested. The knowledge gained from these studies will improve our understanding of plant nematode proteinases and aid the development of a rational proteinase inhibitor-based approach to plant nematode resistance.