Molecular identification of the advanced third-stage larvae (ADV L3) of Gnathostoma lamothei in Tabasco,Mexico

Advanced third-stage larvae (ADV L₃) of Gnathostoma spp. were collected from the muscle tissue of three species of freshwater fish (i.e., Gobiomorus dormitor, Petenia splendida, and Parachromis managuensis) in Swamps of Centla, Tabasco, Mexico. Nine sequences of the ITS2 of the ribosomal DNA of Gnathostoma spp. were compared with sequences obtained from GenBank for G. binucleatum, G. lamothei, G. miyazakii, G. spinigerum, and G. turgidum. Sequences of the ADV L₃ from P. splendida (Isla Chinal), P. managuensis (Isla Chinal), and of two of the six larvae collected from G. dormitor (Tres Brazos), were identical to that of G. binucleatum (GenBank). Sequences from the other four larvae from G. dormitor (Tres Brazos) are identical to the sequence of G. lamothei (GenBank). This is the first record of the intermediate host of G. lamothei. The only species documented to cause human gnathostomiasis in the Americas is G. binucleatum. Our finding of G. binucleatum, and G. lamothei parasitizing the commercially important fish species, G. dormitor in Centla swamps, indicates the possibility of G. lamothei causing human gnathostomiasis in Mexico as well.     

Uncovering signatures of selection in the soybean genome using SSR diversity near QTLs of agronomic importance

The cultivated soybean [Glycine max (L.) Merr.] is widely considered to descend from the wild soybean (G. soja Sieb. & Zucc.). This study was designed to evaluate the genetic variability and differentiation between G. soja and G. max, and to detect signatures of the selection that may have occurred during the domestication process from G. soja to G. max. A total of 192 G. soja accessions and 104 G. max accessions were genotyped using eight selected simple sequence repeat (SSR) markers assigned to three SSR groups. Four SSRs in group A were not located near any known QTL. Three SSRs in group B were associated with seed protein content, and an SSR in group C was associated with resistance to Sclerotinia stem rot. The number of alleles per locus and the level of genetic variability in G. soja were higher than those in G. max. A total of 122 out of 125 alleles were present in G. soja, but only 59 alleles were detected in G. max. The average gene diversity was 0.74 in G. soja and 0.64 in G. max. Four SSRs near QTLs of agronomic importance showed strong genetic differentiation and shift change in high frequency alleles in groups B and C between G. soja and G. max, revealing selection signatures that may reflect the domestication events and recent selective breeding. With reduced diversity in G. max, some undomesticated genes from G. soja should be prime candidates for introgression to increase the pool of diversity in G. max.     

MicroRNA loci support conspecificity of Gyrodactylus salaris and Gyrodactylus thymalli (Platyhelminthes: Monogenea)

The monogenean flatworm Gyrodactylus salaris is a serious threat to wild and farmed Atlantic salmon stocks in Norway. Morphologically, the closely related but harmless Gyrodactylus thymalli on grayling can hardly be distinguished from G. salaris. Until now, molecular approaches could not resolve unambiguously whether G. salaris and G. thymalli represent just one polytypic species, two polytypic species or a complex of more than two species. In the first known genome-wide analysis utilizing 37 conserved microRNA loci, the genetic differentiation of seven populations of G. salaris and G. thymalli was assessed. The concatenated alignment spanned 21,742 bp including 62 variable positions. A neighbor-joining cluster analysis did not support any host-based or mitochondrial haplotype-based grouping of strains. We conclude that a two species concept for G. salaris and G. thymalli does not reflect meaningful biological entities. Instead, G. salaris and G. thymalli are just one species comprising several pathogenic and non-pathogenic strains on various primary hosts. Following the International Code for Zoological Nomenclature, G. salaris Malmberg, 1957 is the valid species name with G. thymalli ?itňan, 1960 becoming the junior synonym. Accordingly, the range of G. salaris is significantly increased, given that formerly G. salaris-free countries such as e.g., Great Britain are now within the species’ natural range. The synonymization of G. salaris and G. thymalli implies severe challenges to current disease management routines, which assume that G. salaris and G. thymalli are readily distinguishable. Protocols for reliable identification of pathogenic and non-pathogenic strains of G. salaris need to be developed.     

Revision and affinities ofGalium (Rubiaceae) in Madagascar

The only representatives ofRubiaceae-Rubieae in Madagascar are five species ofGalium. G. thunbergianum andG. chloroionanthum are essentially afromontane species, ± widely distributed in both the African mainland and Madagascar. The new speciesG. andringitrense andG. ankaratrense are endemic to Madagascar but show close affinities to two afromontane—afroalpine groups from the African mainland, theG. simense—G. ruwenzoriense andG. glaciale complex respectively.G. polyacanthum (new combination) exhibits certain primitive morphological traits and appears to be an old Madagascan endemic without close allies. New chromosome counts for populations from the African mainland demonstrate thatG. chloroionanthum andG. simense are 4x (x=11). Distribution patterns and origins of theGalium species in Madagascar and their relationships with African taxa are discussed.     

A new species of Grillotia Guiart, 1927 (Cestoda: Trypanorhyncha) with redescriptions of congeners and new synonyms

A new species of Grillotia, G. gastrica n. sp., is described from the stomach musculature of the teleosts Upeneichthys lineatus (Bloch & Schneider) and Sillaginodes punctatus (Cuvier) from off Perth, Western Australia. The new species most closely resembles G. pristiophori Beveridge & Campbell, 2001 in having six hooks in each principal row of the metabasal tentacular armature but differs in having a smooth scolex tegument and in having a band of hooklets running the entire length of the external surface of the tentacle rather than diminishing in width to a single hooklet as occurs in G. pristiophori. Grillotia heptanchi (Vaullegeard, 1899) is redescribed and the details of the mature segment are described for the first time. Grillotia adenoplusius (Pintner, 1903) is redescribed from the type-specimens and is considered to be the larval stage of G. acanthoscolex Rees, 1944 (syns G. spinosissima Dollfus, 1969 and G. microthrix Dollfus, 1969). The adult of G. adenoplusius is also redescribed based on the types of G. spinosissima. The type-specimens of G. dolichocephala Guiart, 1935 and G. minor Guiart, 1935 were re-examined and G. minor is considered to be a synonym of G. dolichocephala as is G. meteori Palm & Schröder, 2001. Based on an examination of the type-specimens, G. scolecina (Rudolphi, 1819) is treated as a species inquirenda. A list is provided of the species currently placed in Grillotia.     

Incipient Genome Differentiation in Gossypium. II. Comparison of 12 Chromosomes in G. HIRSUTUM, G. MUSTELINUM and G. TOMENTOSUM Using Heterozygous Translocations

Eight homozygous translocation lines (TT) of G. hirsutum marking 3 chromosomes of the A genome and 9 chromosomes of the D genome were crossed with G. hirsutum, G. mustelinum and G. tomentosum, all homozygous for the standard end arrangements (tt). Chiasma frequencies in the G. hirsutum Tt controls were compared with those in the G. hirsutum x G. mustelinum and the G. hirsutum x G. tomentosum Tt hybrids. Both nucleus-wide and region-specific chiasma frequencies were compared.—Some genome differentiation appears to have arisen between G. hirsutum and G. mustelinum. The G. hirsutum x G. mustelinum hybrids had a 1.8 to 1.9% reduction in the nucleus-wide chiasma frequency. Four of the eight TT lines showed a 3.4 to 10.5% reduction in chiasmata in the hybrid translocation quadrivalents, suggesting that chromosomes 1, 21, 23 and 24 may have undergone localized genome differentiation. The two species may differ naturally in the end arrangement of two chromosomes, since a quadrivalent not due to experimentally introduced translocations was observed in 13% of the PMC's of two G. hirsutum x G. mustelinum hybrids.—Very little genome differentiation has occurred between G. hirsutum and G. tomentosum. In the G. hirsutum x G. tomentosum hybrids, the nucleus-wide estimates showed only a very small (0.1 to 0.2%), though statistically significant, lowering of the chiasma frequency, and there was no reduction in chiasma frequency in the more sensitive readings for specific translocation quadrivalents.     

Protein markers for identification of different species and varieties of cotton

Reference electrophoretic spectra that allow compiling electrophoretic formulas of certain cotton species and varieties were obtained on the basis of analysis of the electrophoretic spectrum of water-soluble and barely soluble proteins of seeds of diploid cotton species of genomic group A (Gossypium arboreum var. indicum, G. arboreum ssp. obtusifolum, G. herbaceum ssp. africanum, and G. herbaceum Harga), group C (G. australe, G. bickii, G. nelsone, and G. sturtianum), group D (G. davidsonii, G. harknessii, G. klotzschianum, G. raimondii, G. thurberi, and G. trilobum), and amphidiploid species of group AD (G. mustelinum, G. hirsutum ssp. palmeri, G. tricuspidatum Bagota, G. tricuspidatum Mari Galanta, G. barbadense L., and G. hirsutum L.)     

Population studies in Goodyera (Orchidaceae) with emphasis on the hybrid origin of G. Tesselata

Morphological, cytological, and paper Chromatographic studies of populations from northern Michigan and examination of herbarium specimens from throughout North America were used to clarify the relationships ofGoodyera oblongifolia, G. repens var.ophioides, andG. tesselata. A canonical analysis of morphological data from mixed populations of these three species depictsG. tesselata as intermediate betweenG. oblongifolia andG. repens var.ophioides. The latter two species are diploid (2n = 30) andG. tesselata is tetraploid (2n = 60). Triploids (2n = ca. 45) were found in two mixed-species populations in northern Michigan.Goodyera tesselata produces three phenolic compounds present inG. oblongifolia and five different compounds present inG. repens var.ophioides. The range ofG. tesselata is confined to glaciated territory (except for two stations) in northeastern North America where the postglacially produced ranges ofG. oblongifolia andG. repens var.ophioides overlap. However,G. tesselata is quite abundant in areas outside the region of sympatry of the other two species. Based on this evidence, it is postulated thatG. tesselata is an allotetraploid species which resulted from hybridization betweenG. oblongifolia andG. repens var.ophioides during early post-Pleistocene. The slightly earlier blooming season ofG. tesselata may have been selected for to provide a measure of reproductive isolation between the tetraploid and its parents and to adapt the new species to the rather short growing season of northeastern North America.     

Biostratigraphy and histories of upper Miocene - Pliocene Globorotalia, South Atlantic and South West Pacific

The upper Miocene-Pliocene histories of the Globorotalia miozea Plexus and G. crassaformis from the S.W. Pacific and a S. Atlantic site are assessed for their biostratigraphic utility in the mid latitudes of the Southern Hemisphere. As in the S.W. Pacific, Pliocene descendants of G. conoidea (G. pliozea, G. mons) appear in the S. Atlantic near the level at which the G. inflata lineage diverges (as G. puncticulata sphericomiozea). But there is an incomplete history of the latter at the Atlantic site examined, probably due to its northern position. Unlike S.W. Pacific sequences, S. Atlantic strata record a phyletic connection between G. juanai (= G. cibaoensis of authors) and G. crassaformis. The alignment of events in the plexus between the two regions is poor as judged by other foraminiferal datums (extinction of Globoquadrina dehiscens, entries of Globorotalia margaritae and G. crassaformis. As with the G. miozea plexus the records of these taxa probably vary according to latitudinal position.     

Phylogenetic placement of Geastrum melanocephalum and polyphyly of Geastrum triplex

Geastrum melanocephalum, originally described as Trichaster melanocephalus, is characterized by large basidiomata and an evanescent endoperidium. Although Trichaster was recently treated as a synonym of Geastrum and the specific name G. melanocephalum has often been used, it is still controversial whether Trichaster is an independent genus. Although a close affinity of G. melanocephalum and G. triplex has been suggested based on some morphological similarities, it is highly likely that G. triplex is polyphyletic because of its high morphological variability. To clarify the phylogenetic position of G. melanocephalum, it is therefore critical to evaluate the monophyly of G. triplex. This study sampled ITS, LSU, and atp6 genes from 144 specimens of Geastrales including G. melanocephalum and G. triplex from several continents. Results of phylogenetic analyses demonstrated G. melanocephalum is nested within Geastrum and is most closely related to the European and North American group of G. triplex. Morphological similarities of G. melanocephalum and European and North American G. triplex are also suggested. Based on phylogenetic and morphological evidence, we confirm Trichaster is a synonym of Geastrum, and the scientific name Geastrum melanocephalum should be accepted. Moreover, the present study revealed that taxa tentatively identified as “G. triplex” are highly polyphyletic, and a taxonomic revision of “G. triplex” is therefore needed.     

Hybridization drives speciation in Gagea (Liliaceae)

Hybridization seems to play an important role in speciation of Gagea Salisb., a genus which is characterised by polyploid taxa lines and in which diploids (2n = 24) appear only to be common in basal sections. Hybrid detection was applied utilising direct and cloning nrDNA ITS data (ITS1, 5.8S rDNA, ITS2) combined with neighbour and ribotype networks and discussed in connection with previously published cpDNA, morphological and karyological data of the authors. We have evidence of the hybrid origin of taxa within the section Gagea (G. pomeranica, G. megapolitana) and the monophyletic clade of sections Didymobulbos and Fistulosae (G. microfistulosa, G. polidorii, G. cf. bohemica). Morphologically and karyologically differentiated Gagea megapolitana and G. pomeranica, adapted to synanthropic habitats, represent both hybrids of G. pratensis × G. lutea. Gagea microfistulosa represents a hybrid of G. villosa × G. fragifera; Gagea polidorii could represent the reverse hybrid. G. glacialis is also closely related to the latter complex.     

In vitro morphogenetic studies on three apomictic species of Garcinia

Comparison of morphogenetic potential of three important Indian species of Garcinia??G. indica, G. cambogia and G. xanthochymus has been reported. Apomictic seeds of G. indica were found to be morphogenetically most potential followed by G. cambogia. The explants of G. xanthochymus were highly recalcitrant towards in vitro conditions and failed to induce adventitious buds on any of the media tested. High frequency direct shoot bud differentiation was induced in aseptic seed cultures of G. indica and G. cambogia on MS medium supplemented with cytokinins (BAP, kinetin or TDZ). Amongst the three cytokinins tested, TDZ (0.1?C0.5???M) was most effective for adventitious bud differentiation in both G. indica and G. cambogia, however, the proliferating buds failed to elongate. Substantial number of buds induced on BAP supplemented media elongated into shoots after subculture on elongation medium. Addition of NAA along with cytokinins in the induction medium enhanced callusing without improvement in bud induction response. The induced adventitious buds were elongated on MS basal medium containing 0.2% activated charcoal. Direct rooting was achieved in both G. indica and G. cambogia on auxin supplemented media with best response at 10???M IBA concentration in both the species. The in vitro raised plantlets showed 90% survival in the field when transferred after hardening and acclimatization.     

Resistant Germplasm in Gossypium Species and Related Plants to Rotylenchulus reniformis

Gossypium hirsutum, G. herbaceum, G. arboreum, G. barbadense, wild Gossypium spp., Hibiscus spp, and other Malvaceae were tested in the greenhouse to identify germplasm resistant to Rotylenchulus reniformis (Rr). Host resistance was based on Rr egg production per gram of root compared with known G. hirsutum susceptible ''Deltapine 16'' as check. G. longicalyx and Sida rhombifolia were nonhosts. High levels of resistance were found in G. stocksii, G. somalense, and G. barbadense ''Texas 110.'' Other cotton lines with potential value in breeding for Rr resistance were G. herbaceum P.I. 408775; G. arboreum P.I. 41895, P.I, 417891, CB 3839; and G. hirsutum 893. All these supported less than 20% of the egg production on the check. Seventy-three percent of the Hibiscus spp. tested were resistant. Female development and egg production reflected host resistance; healthy females and large egg masses were observed on susceptible plants, and degenerated females and small egg masses on resistant plants. Females penetrating nonhost G. longicalyx never matured to kidney shape.     

Xanthones,flavones and secoiridoids of american Gentiana species

Isoorientin, isoorientin-4′-O-glucoside, gentiopicrin, xanthones with 1,3,5,8 or 1,3,7,8-oxidation pattern have been isolated from five North American Gentiana species (G. affinis, G. algida, G. calycosa, G. detonsa, G. strictiflora) and one South American species (G. cerastioides). Separations were achieved with droplet counter-current chromatography (DCCC) or centrifugally accelerated preparative TLC. The distribution of the isolated compounds within the genus is discussed.     

In-depth characterization of the GOTCAB saponins in seven cultivated Gypsophila L. species (Caryophyllaceae) by liquid chromatography coupled with quadrupole-Orbitrap mass spectrometer

Roots of Gypsophila L. (Caryophyllaceae) have been shown to accumulate bidesmosides of triterpenoid carboxylic acids, also called GOTCAB saponins (Glucuronide Oleanane-type Triterpenoid Carboxylic Acid 3, 28-Bidesmosides). The study aimed at in-depth characterization of GOTCABs from root extracts of cultivated Gypsophila scorzonerifolia Ser., G. acutifolia Stev. ex Spreng., G. altissima L., G. pacifica Kom., G. paniculata L., G. oldhamiana Miq. and G. zhegualensis Krasnova using ultra high-performance liquid chromatography coupled with hybrid quadrupol-Orbitrap high resolution mass spectrometry (UHPLC-HRMS). Based on the accurate mass measurements, elemental composition, isotopic peak profiles, fragmentation pattern in tandem mass spectrometry (MS/MS) and literature data, a total of 53 GOTCABs were tentatively identified. In addition, 29 core structures, forming between 2 and 12 isobaric isomers were described. They possess gypsogenin, quillaic and gypsogenic acid as sapogenin, substituted at C-3 with O-β-d-galactopyranosyl-(1 → 2)-[pentosyl-(1 → 3)]-β-d-glucuronopyranoside (β-chain). According to the C-28 ester-bonded oligosaccharide (α-chain) saponins were classified into four groups: GOTCABs with C-28 tetra- and pentasaccharide (type I), GOTCABs with C-28 oligosaccharide substituted with methoxycinnamoyl group (type II), GOTCABs with mono- and diacetylated C-28 oligosaccharide (type III) and GOTCABs with C-28 oligosaccharide substituted with both acetyl and methoxycinamoyl groups (type IV). The possible fragmentation pathways of saponins were proposed. Eleven core structures forming between 2 and 7 isobars are undescribed in the literature. To examine the differences between the assayed Gypsophila species at the same environmental conditions, the variation of saponins was estimated by hierarchical clustering on isobaric fingerprints of GOTCABs. The clustering of the studied species revealed three well-defined clusters. The first cluster comprises G. scorzonerifolia (G1) and G. altissima (G3), characterized by GOTCABs from type III. G. acutifolia (G2) and G. pacifica (G4) formed the second cluster accumulating saponins from types II and III. The third cluster grouped G. paniculata (G5), G. oldhamiana (G6) and G. zhegualensis (G7) sharing GOTCABs from types IV in addition to II and III. This is the first report on the saponins from G. scorzonerifolia and G. zhegualensis. An in-depth depiction of the GOTCAB saponin composition of seven cultivated Gypsophila species was achieved. Therefore, saponins are worth investigating for better understanding of the potential use of Gypsophila roots for pharmaceutical purposes.     

Sex determination in ten crane species by DNA marker EE0.6

Using a unique DNA sequence of W-chromosome EE0.6, we carried out molecular sex determination in 383 individuals of ten species of cranes (Grus grus L., G. leucogeranus Pallas, G. japonensis Muller, G. vipio Pallas, G. canadensis L., G. antigone L., G. monacha Temminck, Anthropoides virgo L., Balearica regulorum Bennett, and B. pavonia L.) kept in zoos and other centers of captive breeding. In 211 birds, sex was determined or verified for the first time. The efficiency of using the sex marker EE0.6 for chicks and immature and adult cranes of different species, as well as for interspecific hybrids was shown.     

An unexpectedly lichenase-stable hexasaccharide from cereal,horsetail and lichen mixed-linkage β-glucans (MLGs): Implications for MLG subunit distribution

Mixed-linkage (1→3),(1→4)-β-d-glucan (MLG) is a biologically and technologically important hemicellulose, known to occur in three widely separated lineages: the Poales (including grasses and cereals), Equisetum (fern-allies), and some lichens e.g. Iceland moss (Cetraria islandica). Lichenase (E.C. 3.2.1.73) is widely assumed to hydrolyse all (1→4) bonds that immediately follow (1→3) bonds in MLG, generating predominantly the tetrasaccharide β-d-Glcp-(1→4)-β-d-Glcp-(1→4)-β-d-Glcp-(1→3)-d-Glc (G4G4G3G; MLG4), the corresponding trisaccharide (G4G3G; MLG3), and sometimes also laminaribiose (G3G; MLG2). The ratio of the oligosaccharides produced characterises each polysaccharide. We report here that digestion of MLG from barley (Hordeum vulgare), Equisetum arvense and C. islandica by Bacillus subtilis lichenase also yields the unexpectedly stable hexasaccharide, β-d-Glcp-(1→3)-β-d-Glcp-(1→4)-β-d-Glcp-(1→4)-β-d-Glcp-(1→4)-β-d-Glcp-(1→3)-d-Glc (G3G4G4G4G3G, i.e. MLG2–MLG4), identified by thin-layer chromatography, gel-permeation chromatography, HPLC (HPAEC), β-glucosidase digestion, ¹H/¹³C-NMR spectroscopy and mass spectrometry. On HPLC, G3G4G4G4G3G is the major constituent of a peak previously ascribed solely to the nonasaccharide G4G4G4G4G4G4G4G3G. Because it was widely presumed that lichenase would cleave G3G4G4G4G3G to MLG2 + MLG4, our data both redefine the substrate specificity of Bacillus lichenase and show previous attempts to characterise MLGs by HPLC of lichenase-digests to be flawed. MLG2 subunits are particularly underestimated; often reported as negligible, they are here shown to be an appreciable constituent of MLGs from all three lineages. We also show that there is no appreciable yield of water-soluble lichenase products with DP > 9; potential identities of products previously labelled DP > 9 are suggested. Finally, this discovery also provides a opportunity to investigate the spatial distribution of subunits along the MLG chain. We show that MLG2 subunits in barley and Cetraria MLG are not randomly distributed, but predominantly found at the non-reducing end of MLG4 subunits.     

Order of enzymic incorporation of O-methyl groups into the O-methyl-d-glucose-containing polysaccharide of Mycobacterium smegmatis. A tritium-labelling study

The order of enzymic incorporation of O-methyl groups into the O-methyl-d-glucose-containing polysaccharide (MGP) of Mycobacterium smegmatis, 3MG(J)→G(I)→G(H)→G(G)→6MG(F)→(GMG)₉(E)→[G(L)→G(D)]→G(C)→[G(K)→G(B)]→G(A)→Ga, where G is d-glucose, 3MG is 3-O-methyl-d-glucose, 6MG is 6-O-methyl-d-glucose, and Ga is d-glyceric acid, was studied by incubating cultures of M. smegmatis with l-[³H-Me]methionine for various times. MGP was then extracted from the cells, and relative radioactivities of residues D, (E + F)_average and J, or of D, E_average, F, and J, were determined. Tritium-labelling of these residues increased in the reducing-to-nonreducing residue direction, the steepness of the gradient becoming more shallow with increasing incubation time. The results are consistent with a biosynthetic mechanism that involves sequential addition of O-methyl groups to residues of the pre-formed d-glucan, in the reducing-to-non-reducing residue direction.     

Three new species of galium (rubiaceae) from Baja California

The species described as new are apparently narrow endemics, all belonging to that section ofGalium having fruits with long straight specialized hairs.Galium wigginsii andG. moranii are of the widespreadG. wrightii-G. parishii complex, whereasG. carterae is most like the rarely collectedG. oresbium Greenman of northeastern Mexico.