Descriptions of Three New Longidorus Species from Alaska (Nematoda: Longidoridae)

Three new Longidorus species, L. alaskaensis n. sp., L. paralaskaensis n. sp., and L. bernardi n. sp., are described from specimens collected near Fairbanks, Alaska. Longidorus alaskaensis differs from all species of Longidorus by the presence of a caecum-like structure situated at the reflex of the oviduct. Longidorus paralaskaensis most closely resembles L. alaskaensis n. sp., L. crassus Thorne, L. picenus Roca, Lamberti &Agostinelli, and L. silvae Roca, differing from the last three of these species by having a parallel vs. a tapered lip region, and from all four by having a more narrowly rounded tail tip. Longidorus paralaskaensis differs from L. alaskaensis by having a longer odontostyle (119-128 vs. 110-118 μm) and by lacking the caecum-like structure found at the reflex of the oviduct. Longidorus bernardi n. sp. most closely resembles L. mirus Khan, Chawla &Seshadri, from which it differs by having a longer tail with a more acutely rounded tip, a longer body length (3.5-4.6 vs. 3.0-3.6 μm), and a larger c'' value (1.6-1.8 vs. 1.3-1.6). Longidorus bernardi differs from L. sylphus Thorne, L. africanus Merny, L. auratus Jacobs &Heyns, and L. conicaudatus Khan by having a slightly expanded lip region vs. a lip region with parallel body walls and a more finely rounded tail tip.     

Phylogenic analysis of the genus Leishmania by cytochrome b gene sequencing

In a previous report (Luyo-Acero et al., 2004), we demonstrated that cytochrome b (Cyt b) gene analysis is an effective method for classifying several isolates of the genus Leishmania; hence, we have further applied this method to other Leishmania species in an effort to enhance the accuracy of the procedure and to construct a new phylogenic tree. In this study, a total of 30 Leishmania and Endotrypanum WHO reference strains, clinical isolates from our patients assigned to 28 strains (human and non-human pathogenic species) and two species of the genus Endotrypanum were analyzed. The Cyt b gene in each sample was amplified by PCR, and was then sequenced by several primers, as reported previously. The phylogenic tree was constructed based on the results obtained by the computer software MEGA v3.1 and PAUP* v4.0 Beta. The present phylogenic tree was almost identical to the traditional method of classification proposed by Lainson and Shaw (1987). However, it produces the following suggestions: (1) exclusion of L. (Leishmania) major from the L. (L.) tropica complex; (2) placement of L.tarentolae in the genus Sauroleishmania; (3) L. (L.) hertigi complex and L. (V.) equatorensis close to the genus Endotrypanum; (4) L. (L.) enrietti, defined as L. (L.) mexicana complex, placed in another position; and (5) L. (L.) turanica and L. (L.) arabica are located in an area far from human pathogenic Leishmania strains. Cyt b gene analysis is thus applicable to the analyzing phylogeny of the genus Leishmania and may be useful for separating non-human pathogenic species from human pathogenic species.     

Pollen morphology of Lathyrus (Fabaceae) taxa in the Platystylis section from Turkey

In this study, the pollen morphology of 18 wild taxa of Lathyrus L. grown in Turkey: L. pallescens (Bieb.) Koch, L. brachypterus Cel., L. haussknechtii Sirj., L. karsianus P. H. Davis, L. satdaghensis P. H. Davis, L. nivalis Hand.-Mazz, L. atropatanus (Grossh.) Sir., L. armenus (Boiss. and Huet) Sirj., L. cyaneus (Stev.) Koch var. cyaneus, L. cyaneus var. pinnatus?Davis, L. digitatus (Bieb.) Fiori, L. tukhtensis Czecz., L. variabilis (Boiss. and Ky.) Maly, L. spathulatus Cel. L. elongatus (Bornm.) Sirj., L. cilicicus Hayek and Siehe, L. boissieri Sirj., and L. bitlisicus Pe?men was examined by light microscopy and scanning electron microscopy. The pollen grains were 3-zonocolporate, of spheroidal-subprolate-prolate (P/E?=?0.957?C1.252) types, and medium in size. Equatorial view: rectangular or elliptical-obtuse-convex, polar view: circular, triangular or quinquangular-obtuse-convex. The smallest pollen grains belonged to L. elongatus (P?=?36.972/E?=?38.636) and the largest to L. cyaneus var. cyaneus (P?=?46.332/E?=?32.864). The ornamentation was perforate-foveolate or slightly reticulate. Some photographs included in this work were taken using both light microscopy and scanning electron microscopy.     

Three new species of Liparis s.l. (Orchidaceae: Malaxideae) from Southwest China based on morphological characters and phylogenetic evidence

Liparis aureolabella and L. mengziensis, two new species from the karst region of southwestern China, and L. bingzhongluoensis, a new species from montane region in Yunnan, are described and illustrated. L. aureolabella is easily distinguished from its relatives by having abaxially purple leave with purple reticulate veins prominent adaxially, a lip auriculate at base, and falcate-lanceolate pollinia. Liparis mengziensis is closely related to L. petiolata and L. auriculata, but differs from them by having an ovate to broadly ovate leaf, purple lip and apex connate along the margins. Liparis bingzhongluoensis is similar to Liparis nanlingensis, but the new species is characterized by having a lip with two transparent ridges on its disc, longitudinally concave basal callus and triangular column wings. Phylogenetic analyses based on nuclear ribosomal ITS and plastid matK sequences showed that L. aureolabella and L. mengziensis are nested with L. petiolata or L. auriculata in a monophyletic clade. L. bingzhongluoensis is sister to a clade formed by L. nanlingensis, L. tsii, L. sasakii and L. krameri. Moreover, morphological comparisons strongly support that the three species as separated species newly to science.     

Leishmania species: Detection and identification by nested PCR assay from skin samples of rodent reservoirs

Many rodent species act as reservoir hosts of zoonotic cutaneous leishmaniasis in endemic areas. In the present study a simple and reliable assay based on nested PCR was developed for the detection and identification of Leishmania parasites from rodent skin samples. We designed Leishmania-specific primers that successfully amplified ITS regions of Leishmania major, Leishmania gerbilli and Leishmania turanica using nested PCR. Out of 95 field collected Rhombomys opimus, 21 were positive by microscopic examination and 48 by nested PCR. The percentage of gerbils infected with L. major, L. gerbilli and L. turanica was 3.2%, 1.1% and 27.4%, respectively. In 15.8% of the rodents, we found mixed natural infections by L. major and L. turanica, 1.1% by L. major and L. gerbilli, and 2.1% by the three species. We concluded that this method is simple and reliable for detecting and identifying Leishmania species circulating in rodent populations.     

Analysis of MC1R genetic variation in Lepus species in Mediterranean refugia

A study on the inter- and intraspecies variation of MC1R gene was performed in Lepus species inhabiting the Mediterranean basin (L. granatansis, L. europaeus, L. corsicanus, L. castroviejoi and L. mediterraneus) and their neighboring species in Europe (L. timidus) and Africa (L. saxatilis, L. capensis), in order to infer micro- versus macroevolutionary adaptation. Eleven different sequences were isolated that corresponded to five amino acid sequences. Comparison of MC1R nucleotide phylogenetic tree with phylogenies resulting from mtDNA regions of the same species showed absence of congruence between these sets of markers. The Mediterranean area that offered refugia during last glaciation retains more MC1R genotypes compared with populations of North and Central Europe as a consequence of founder effects. L. corsicanus and L. castroviejoi bore identical alleles supportive of their conspecificity, as indicated by other molecular markers. Within L. europaeus, a group of Israeli hares were distinguished by a different MC1R functional allele; additional differences in coat colour and other genetic markers raise doubts about its taxonomic status. Finally, the present data reinforced the idea of bi-directional introgressive hybridization between L. europaeus and L. timidus in Switzerland.     

Patterns in Nuclear and Mitochondrial DNA Reveal Historical and Recent Isolation in the Black-Tailed Godwit (Limosa limosa)

On the basis of morphological differences, three subspecies of Black-tailed Godwit (Limosa limosa) have been recognized (L. l. limosa, L. l. islandica and L. l. melanuroides). In previous studies mitochondrial DNA (mtDNA) sequence data showed minimal genetic divergence between the three subspecies and an absence of sub-structuring within L. l. limosa. Here, population genetic structure and phylogeographic patterns have been analyzed using COI, HVR1 and HVR2 mtDNA sequence data as well as 12 microsatellite loci (nuDNA). The nuDNA data suggest genetic differentiation between L. l. limosa from Sweden and The Netherlands, between L. l. limosa and L. l. islandica, but not between L. l. limosa and L. l. melanuroides. However, the mtDNA data were not consistent with the nuDNA pattern. mtDNA did support a split between L. l. melanuroides and L. l. limosa/L. l. islandica and also demonstrated two L. l. limosa haplotype clusters that were not geographically isolated. This genetic structure can be explained by a scenario of isolation of L. l. melanuroides from L. l. limosa in Beringia during the Last Glacial Maximum. During the Pleistocene separation of L. l. islandica from L. l. limosa occurred, followed by colonization of Iceland by the L. l. islandica during the Holocene. Within L. l. limosa founder events, followed by population expansion, took place during the Holocene also. According to the patterns observed in both markers together and their geographic separation, we propose that the three traditional subspecies indeed represent three separate genetic units.     

Pollen morphology of Lathyrus (Leguminosae) taxa belonging to Lathyrus, Orobastrum and Cicercula sections from Turkey

The pollen morphology of 20 wild taxa belonging to Lathyrus (Syn: Eulathyrus), Orobastrum (Taub.) Boiss. and Cicercula (Medic.) Gren. & Godr. sections of Lathyrus L. grown in Turkey (L. rotundifolius Wild. subsp. miniatus (Bieb. ex Steven) P.H. Davis, L. grandiflorus Sibth. & Sm., L. saxatilis (Vent.) Vis., L. vinealis Boiss. & No?, L. inconspicuus L. var. inconspicuus, L. inconspicuus var. stenophyllus (Boiss.) Rech. f., L. tauricola P.H. Davis, L. woronowii Bornm., L. hierosolymitanus Boiss., L. cassius Boiss., L. gorgoni Parl. var. gorgoni, L. pseudo-cicera Pamp., L. sativus L., L. blepharicarpus Boiss., L. stenophyllus Boiss. & Heldr., L. belinensis Maxted & Goyder, L. phaselitanus Hub.-Mor. & P.H.Davis, L. chrysanthus Boiss., L. chloranthus Boiss., and L. trachycarpus (Boiss.) Boiss were examined by light microscopy (LM) and scanning electron microscopy (SEM) in this study. The pollen grains were 3-zonocolporate, spheroidal, subprolate, and prolate (P/E?=?0.99–1.48) types, and were medium in size (equatorial view: rectangular or elliptical-obtuse-convex; polar view: circular, triangular and quinquangular-obtuse-convex). The smallest pollen grains belonged to L. tauricola (P?=?30.94/E?=?31.20) and the largest to L. grandiflorus (P?=?50.60/E?=?36.40). The ornamentation was reticulate and reticulate-granulate in the mesocolpium, and usually psilate in the apocolpium. Some photographs included in this work were taken using both LM and SEM.     

RAPD analysis of genome polymorphism in the family Lemnaceae

The multilocus RAPD analysis of intergeneric, inter-and intraspecific nuclear genome polymorphism was used for the first time to assess intergeneric, interspecific, and intraspecific polymorphism in Lemnaceae growing on the territory of Russia. The origin of the chosen accessions overlapped with the natural range of duckweeds in Russia. Seventy-five Lemnaceae accessions representing eight species (L. minor, L. gibba, L. turionifera, L. japonica, L. trisulca, L. aequinoctialis, S. polyrhiza, and L. punctata) from three genera (Lemna, Spirodela, and Landoltia), were analyzed. The highest variability levels were revealed in L. minor accessions (0.03–0.20). Species L. trisulca and S. polyrhiza were characterized by values of genetic distance 0.01–0.18 and 0.03–0.16, respectively. The lowest polymorphism levels were detected for L. turionifera (0.01–0.11). The dendrogram based on RAPD data showed that L. aequinoctialis was the most genetically distant species of the genus Lemna. Accessions of species L. turionifera and L. japonica, as well as L. minor and L. gibba, did not form separate species-specific subclusters; rather, they fell into clusters with L. japonica/L. turionifera and L. minor/L. gibba. Accessions of the genera Spirodela and Landoltia formed two separate clusters combined into one group.     

Descriptions of Three New Longidorus Species from Slovakia (Nemata: Longidoridae)

Three new Longidorus species from Slovakia are described. Longidorus carpathicus n. sp. most closely resembles Longidorus silvae but differs by having a longer odontostyle, odontophore, and total stylet; smaller a and c ratios; and longer distance to the guide ring. This new species also resembles L. picenus, L. macrosoma, and L. major but differs by having a narrower lip width. It further differs from L. picenus by having a longer odontostyle and smaller c ratio, and by lacking males; from L. macrosoma by having a longer odontostyle, smaller c ratio, by lacking males, and a more pronounced J1 tail peg; and from L. major by having a shorter body length, longer odontostyle, longer odontophore, and longer J1 tail peg. Longidorus piceicola n. sp. most closely resembles L. eridanicus, from which it differs by having a greater lip width, longer tail, smaller c ratio, larger c'' ratio, shorter hyaline tail length, and a conically rounded vs. hemispherical tail. This new species differs from L. cylindricaudatus by having a larger lip width, longer odontostyle and odontophore, and a greater distance to the guide ring; from L. nevesi by having a shorter body length, longer odontostyle, larger c'' ratio, and shorter hyaline tail length. Longidorus juglansicola n. sp. most closely resembles L. athesinus but differs by its longer body, wider lips, and larger a and c ratios. It closely resembles L. vineacola but differs by its shorter body length, smaller c ratio, and an almost parallel lip outline vs. an expanded lip outline; from L. lusitanicus by a longer odontophore and tail, and an almost parallel lip outline vs. an expanded lip outline.     

Taxonomical studies on Oriental Platygastridae (Hymenoptera: Platygastroidea)

14 species new to science are described, viz. Amblyaspis joenssoni, Euxestonotus sabahensis, Leptacis cheyi (all from Malaysia), L. jani (from Laos), L. maliauensis (from Malaysia), L. ongkudoni (from Malaysia), L. pederseni (from Laos), L. reticulaticeps (from Malaysia), L. solodovnikovi, L. vilhelmseni (both from Laos), Sacespalus viklundi (from Malaysia), Synopeas laosianum (from Laos), S. opaciceps and S. waidii (both from Malaysia). The following species described in Leptacis by Ushakumari, R., Narendran, T.C., 2007. A taxonomic revision of Leptacis Foerster (Hymenoptera: Platygasteridae) of India. Rec. Zool. Surv. India 107, 7–32 are transferred to Synopeas: L. aeros, L. alus, L. asiaticus, L. benazeer, L. diversus, L. manii, L. mustus, L. nuperus and L. scaposus. Synopeas saltaense is a nom. nov. for S. intermedius Buhl, 2005 preoccupied by S. intermedius (Ushakumari, R., 2004. Diversity of Platygaster Latreille (Hymenoptera: Platygastridae) of Kerala. In Rajmohana, K., Narendran, T.C., Perspectives on biosystematics and biodiversity: Prof. T. C. Narendran commemoration volume. Systematic Entomology Research Scholars Association, University of Calicut, Kozhikode, India, pp. 573–591). New locality records for 20 already known Oriental platygastrid species are given.     

Phylogenetic studies in the polyploid complex of the genus Leucanthemum Mill. (Compositae, Anthemideae) based on cpDNA sequence variation

To reconstruct the evolutionary history of the polyploid southern and central European genus Leucanthemum, comprising 41 species with ploidy levels ranging from 2x to 22x, we analysed chloroplast DNA sequence variation (psbA-trnH and trnL-trnF intergenic spacer regions) of 106 representatives belonging to 30 species and 41 taxa. In an unrooted haplotype network, which shows internal (ancestral) haplotypes that are mainly represented by diploid taxa (L. gallaecicum, L. gracilicaule, L. halleri, L. laciniatum, L. lithopolitanicum, L. rotundifolium, and L. tridactylites), we identified three major haplotype groups (HTGs) containing diploid and polyploid species. Whereas HTG?I contains most of the polyploid taxa of the genus, with a single diploid species from the SW Alps (L. virgatum), HTG?II consists of four diploid (L. burnatii, L. gaudinii, L. graminifolium, and L. vulgare) and six polyploid species, and HTG?III comprises one diploid (L. pluriflorum) and three polyploid species endemic to the NW part of the Iberian Peninsula. We also further found evidence for recurrent formation of at least three polyploid taxa (i.e., L. delabrei, L. ircutianum subsp. cantabricum, and L. pallens).     

Floral divergence in closely related Leucospermum tottum (Proteaceae) varieties pollinated by birds and long-proboscid flies

The Proteaceae are renowned for their floral diversity but surprisingly the role of pollinators in driving evolutionary divergence in this family has been underexplored. Here we focus on recently diverged taxa to gain insight into the processes that generate diversity by testing whether two varieties of Leucospermum tottum might have originated by pollinator mediated adaptive divergence. L. tottum var. tottum has pale salmon-coloured horizontally-oriented flowers, long nectar tubes, and small volumes of concentrated nectar. L. tottum var. glabrum has red and yellow vertically oriented flowers, short nectar tubes, and large volumes of dilute nectar. Despite the morphological divergence, the varieties are indistinguishable using eight molecular markers, indicating a very early stage of differentiation. Consistent with their morphologies, L. tottum var. tottum is pollinated by long-proboscid flies (Philoliche rostrata and Philoliche gulosa), Cape sugarbirds (Promerops cafer), and, to a lesser extent, by Orange-breasted sunbirds (Anthobaphes violacea), whereas, L. tottum var. glabrum is pollinated only by Orange-breasted sunbirds. A. violacea visits both varieties, but makes more frequent contact with pollen presenters when foraging on L. tottum var. glabrum. The exclusion of birds caused a steeper reduction in seed production in L. tottum var. glabrum than in L. tottum var. tottum, consistent with specialization for bird-pollination in this variety. Additionally, L. tottum var. glabrum exhibits autogamy, whereas L. tottum var. tottum does not. Floral divergence between the two L. tottum varieties corresponds with divergence in pollinator use.     

A biosystematic study of critical species ofLactuca sect.Lactucopsis

In the present paper the results of a biosystematic study of some critical species ofLactuca sect.Lactucopsis are given. After a morphological, chorological, partly cytotaxonomical, chemotaxonomical analysis and experimental crossings the author came to the conclusion that bothL. wilhelmsiana Fisch. etMey ex. DC. andL. chaixii Vill. should be treated as infraspecific taxa ofL. quercina L. No differences were found betweenL. quercina andL. chaixii except in the basic shape of the cauline leaves.L. wilhelmsiana, however, can be distinguished by the length of the beak of the achene and by the distributional area. After revision the correct names of the taxa studied are as follows:Lactuca quercina L.,L. quercina var.integrifolia Bogenh. (Bisch.) forL. chaixii, L. quercina subsp.wilhelmsiana (Fisch. etMey.ex DC.)Feráková forL. wilhelmsiana     

Structural comparison of lipophosphoglycan from Leishmania turanica and L. major,two species transmitted by Phlebotomus papatasi

The lipophosphoglycan (LPG) of Leishmania major has a major role in the attachment to Phlebotomus papatasi midgut. Here, we investigated the comparative structural features of LPG of L. turanica, another species transmitted by P. papatasi. The mAb WIC 79.3, specific for terminal Gal(β1,3) side-chains, strongly reacted with L. turanica LPG. In contrast, L. turanica LPG was not recognized by arabinose-specific mAb 3F12. In conclusion, LPGs from L. major and L. turanica are similar, with the latter being less arabinosylated than L. major's. The high galactose content in L. turanica LPG is consistent with its predicted recognition by P. papatasi lectin PpGalec.     

Effects of phosphorus supply on growth, phosphate concentration and cluster-root formation in three Lupinus species

Background and Aims

In some lupin species, phosphate deficiency induces cluster-root formation, which enhances P uptake by increasing root surface area and, more importantly, the release of root exudates which enhances P availability.

Methods

Three species of Lupinus, L. albus, L. atlanticus and L. micranthus, with inherently different relative growth rates were cultivated under hydroponics in a greenhouse at four phosphate concentrations (1, 10, 50 and 150 µm) to compare the role of internal P in regulating cluster-root formation.

Key Results

The highest growth rate was observed in L. atlanticus, followed by L. albus and L. micranthus. At 1 µm P, cluster-root formation was markedly induced in all three species. The highest P uptake and accumulation was observed in L. micranthus, followed by L. atlanticus and then L. albus. Inhibition of cluster-root formation was severe at 10 µm P in L. atlanticus, but occurred stepwise with increasing P concentration in the root medium in L. albus.

Conclusions

In L. atlanticus and L. albus cluster-root formation was suppressed by P treatments above 10 µm, indicating a P-inducible regulating system for cluster-root formation, as expected. By contrast, production of cluster roots in L. micranthus, in spite of a high internal P concentration, indicated a lower sensitivity to P status, which allowed P-toxicity symptoms to develop.     

Development of species-specific PCR and PCR-restriction fragment length polymorphism assays for L.infantum/L.donovani discrimination

Discrimination of Leishmaniainfantum and L. donovani, the members of the L. (L.) donovani complex, is important for diagnosis and epidemiological studies of visceral leishmaniasis (VL). We have developed two molecular tools including a restriction fragment length polymorphisms of amplified DNA (PCR-RFLP) and a PCR that are capable to discriminate L. donovani from L. infantum. Typing of the complex was performed by a simple PCR of cysteineproteaseB (cpb) gene followed by digestion with DraIII. The enzyme cuts the 741-bp amplicon of L. donovani into 400 and 341 bp fragments whereas the 702 bp of L. infantum remains intact. The designed PCR species-specific primer pair is specific for L. donovani and is capable of amplifying a 317 bp of 3’ end of cpb gene of L. donovani whereas it does not generate an amplicon for L. infantum. The species-specific primers and the restriction enzyme were designed based on a 39 bp insertion/deletion (indel) in the middle of the cpb gene. Both assays could differentiate correctly the two species and are reliable and high-throughput alternatives for molecular diagnosis and epidemiological studies of VL in various foci.