Scanning Electron Microscopy of Xiphinema,Longidorus, and Californidorus Stylet Morphology

Stylet ultrastructure of five Xiphinema, four Longidorus, and three Californidorus species was compared by scanning electron microscopy. Morphological differences were seen in the odontophores and odontostyle bases between the genera and some of the species. All Xiphinema studied had well-developed odontophore flanges; the Longidorus species lacked flanges, except for weakly developed ones in L. diadecturus; and none of the Californidorus had flanges. Three sinuses were present in the odontophores of all species. The sinuses varied in length depending upon species. In Xiphinema and Californidorus the odontostyle bases had distinct overlapping collars, but in Longidorus the collars were absent except for L. diadecturus. The odontostyle-odontophore junction from a lateral view appeared as a slanted transverse line in all the species, but in a dorsal view of Xiphinema and Californidorus it was V-shaped. Dorsal longitudinal seams of the odontostyle and odontophore were observed in all the species. The dorsally located odontostyle aperture was ca. 1 μm from the anterior end in all species, except in one Longidorus sp. it was ca. 4 μm from the end.     

Phylogenetic Relationships and Genetic Variation in Longidorus and Xiphinema Species (Nematoda: Longidoridae) Using ITS1 Sequences of Nuclear Ribosomal DNA

Genetic analyses using DNA sequences of nuclear ribosomal DNA ITS1 were conducted to determine the extent of genetic variation within and among Longidorus and Xiphinema species. DNA sequences were obtained from samples collected from Arkansas, California and Australia as well as 4 Xiphinema DNA sequences from GenBank. The sequences of the ITS1 region including the 3'' end of the 18S rDNA gene and the 5'' end of the 5.8S rDNA gene ranged from 1020 bp to 1244 bp for the 9 Longidorus species, and from 870 bp to 1354 bp for the 7 Xiphinema species. Nucleotide frequencies were: A = 25.5%, C = 21.0%, G = 26.4%, and T = 27.1%. Genetic variation between the two genera had a maximum divergence of 38.6% between X. chambersi and L. crassus. Genetic variation among Xiphinema species ranged from 3.8% between X. diversicaudatum and X. bakeri to 29.9% between X. chambersi and X. italiae. Within Longidorus, genetic variation ranged from 8.9% between L. crassus and L. grandis to 32.4% between L. fragilis and L. diadecturus. Intraspecific genetic variation in X. americanum sensu lato ranged from 0.3% to 1.9%, while genetic variation in L. diadecturus had 0.8% and L. biformis ranged from 0.6% to 10.9%. Identical sequences were obtained between the two populations of L. grandis, and between the two populations of X. bakeri. Phylogenetic analyses based on the ITS1 DNA sequence data were conducted on each genus separately using both maximum parsimony and maximum likelihood analysis. Among the Longidorus taxa, 4 subgroups are supported: L. grandis, L. crassus, and L. elongatus are in one cluster; L. biformis and L. paralongicaudatus are in a second cluster; L. fragilis and L. breviannulatus are in a third cluster; and L. diadecturus is in a fourth cluster. Among the Xiphinema taxa, 3 subgroups are supported: X. americanum with X. chambersi, X. bakeri with X. diversicaudatum, and X. italiae and X. vuittenezi forming a sister group with X. index. The relationships observed in this study correspond to previous genera and species defined by morphology.     

Longidorus biformis n. sp. and L. glycines n. sp. (Nematoda: Longidoridae): Two Amphimictic Species from Arkansas

Two new amphimictic species of Longidorus were found in Arkansas. Longidorus biformis n. sp., found in the rhizosphere of hardwood trees along streams in sandy soil in 14 Arkansas locations, is characterized by its long body (5.42-9.50 mm), wide expanded flattened head end, head width 20.0 to 26.0 µm, odontostyle 96 to 125 µm, guide ring 29 to 38 µm posterior to the anterior end, elongate conoid tail, and c'' = 0.9-2.1. Females with 2 to 11 vetromedian supplement-like structures were found in 2 of 14 populations of this new species. Longidorus biformis n. sp. is closest to L. seinhorsti Peneva, Loof &Brown, 1998 and L. closelongatus Stoyanov, 1964. Among North American species it is closest to L. glycines n. sp. A distinguishing feature of L. biformis n. sp. is the presence of supplement-like organs in some females. Longidorus glycines n. sp., found in soybean microplots at the Main Research Station, Fayetteville, Arkansas, is characterized by its long body (6.14-8.31 mm), wide offset flattened head end, head width 20.3 to 23.3 µm, odontostyle 87.3 to 99.5 µm, guide ring 22.3 to 26.4 µm posterior to the anterior end, short conoid tail with rounded terminus, and c'' = 0.9-1.4. Longidorus glycines n. sp. is closest to L. lusitanicus Macara, 1985. Among North American species it is close to L. biformis n. sp., L. breviannulatus Norton and Hoffman, 1975, and L. crassus Thorne, 1964. Both new species are believed to have four juvenile stages; the first stage was not found for L. biformis n. sp.     

Tissue culture of forest trees: Clonal multiplication of Eucalyptus grandis L.

Axillary shoot proliferation was obtained using explants of Eucalyptus grandis L. juvenile and mature stages on a defined medium. Murashige and Skoog medium (MS) supplemented with benzyladenine (BA), naphthalene acetic acid (NAA) and additional thiamine. Excised shoots were induced to root on a sequence of three media: (1) White's medium containing indoleacetic acid (IAA), NAA and indole butyric acid; (IBA), (2) half-strength MS medium with charcoal and (3) half-strength MS liquid medium. The two types of explants differed in rooting response, with juvenile-derived shoots giving 60% rooting and adult-derived ones only 35%. Thus, the factors limiting cloning of selected trees in vitro are determined to be those controlling rooting of shoots in E. grandis.     

Stepwise and Canonical Discriminant Analysis of Longidorus Species (Nematoda: Longidoridae) from Arkansas

During a 1998-to-2001 survey from Arkansas, nine distinct species of Longidorus were found including five new species. Morphometrics of these nine species were used in a stepwise and canonical discrimination to select a subset of characteristics that best identified each species. Student''s t test was applied to compare Longidorus breviannulatus Norton &Hoffman, 1975; L. crassus Thorne, 1974; L. diadecturus Eveleigh &Allen, 1982; L. fragilis Thorne, 1974; L. biformis Ye &Robbins, 2004; L. glycines Ye &Robbins, 2004; L. grandis Ye &Robbins, 2003; L. paralongicaudatus Ye &Robbins, 2003; and L. paravineacola Ye &Robbins, 2003 to examine interspecies variation and test for the most useful morphometric characters in species discrimination. Most of the morphometric characters were useful to differentiate species, but species identification could not be based on a single character because the morphometric character ranges often overlap. Stepwise discriminant analysis indicated that the guide ring position, head width, tail length, body length, odontostyle length, and anal body width were the most important variables. These were used to generate canonical variables in discriminating the species. The first three canonical variables accounted for 95% of the total variance. The scatterplots by the first three canonical variables grouped and separated the Longidorus species from Arkansas. Stepwise and canonical discriminant analyses were useful for examining the groupings and morphometric relationships of the nine Longidorus species.     

Barriers to interspecific hybridization between Juglans nigra L. and J. regia L species

Juglans nigra and Juglans regia are phylogenetically divergent species. Despite the economic interest in Juglans?×?intermedia (J. nigra?×? J. regia), walnut hybridization is rare under natural conditions and still difficult using controlled pollination. Here, we evaluated some reproductive mechanisms that may prevent successful natural hybridization. The study of flowering phenology of 11 J. nigra and 50 J. regia trees growing in a plantation provided information regarding the opportunity for interspecific crosses. Variation in flower size, pollen quality of putative donors, and variation in seed yield and rate of hybrid production among putative maternal trees were examined. DNA fingerprinting and parentage analyses based on nine microsatellites permitted the identification of hybrids and hybridogenic parent. Our data indicated that overlap occurred between the staminate flowering of protogynous J. regia and the beginning of pistillate flowering of protogynous J. nigra. Differences in floral size were computed between walnut species. Only three hybrids among 422 offspring of eleven J. nigra progenies were identified. Interspecific hybridization involving pollination of one early-flowering-protogynous J. nigra by three protogynous J. regia trees was detected. The correct development of J. regia male gametophytes, high pollen viability (86.5 %), and germination (57.6 %) ruled out the possibility that low pollen quality contributed to depressed hybrid production. Our findings indicated that these two species tended to remain reproductively isolated. The substantial disjunction in flowering time and additional prezygotic barriers such as differences in floral size and conspecific pollen advance may affect interspecific gene flow between J. regia and J. nigra.     

Interspecific differences in prey items in relation to morphological characteristics among four lutjanid species (Lutjanus decussatus, L. fulviflamma, L. fulvus and L. gibbus)

Clarifying interspecific differences in prey items in relation to morphological characteristics is a fundamental aspect to understand the mechanism enabling the diversity of feeding ecology of fishes. The aim of the present study was to clarify the relationship between prey items and body shape variation, teeth and mandible characteristics for four lutjanid species: Lutjanus decussatus, L. fulviflamma, L. fulvus and L. gibbus. Stomach contents analysis revealed that the main prey items of L. decussatus were fishes, L. fulviflamma were crabs and fishes, and L. fulvus and L. gibbus was crabs. Body shape analysis revealed that L. decussatus and L. fulviflamma had a shallower body depth whereas L. fulvus and L. gibbus had a deeper body depth. The two species with a shallower body had long teeth whereas the other two species with a deeper body depth had shorter teeth. The jaw-lever mechanics were compared and L. decussatus and L. fulviflamma have a faster mouth opening–closing mechanism. Canonical correspondence analysis revealed that fishes was the major prey item for the species having a shallower body depth, higher teeth length and lower ratios of in-lever to out-lever of mandibles, whereas crabs was the major prey item for the species having a deeper body depth, lower teeth length and higher ratios of in-lever to out-lever of mandibles. It is suggested that the interspecific differences in main prey items among the four species are directly related to behavioral differences based on body shape, teeth characteristics and jaw-lever mechanics.     

Molecular Characterization of a Xiphinema hunaniense Population with Morphometric Data of all Four Juvenile Stages

A population of Xiphinema hunaniense Wang and Wu, 1992 with all four juvenile stages was found in the rhizosphere of Pinus sp. in Hangzhou, Zhejiang, China. Morphometrics of 18 females and 35 juveniles of this population are given herein. Detailed morphology and morphometrics of the four juvenile stages are provided. Further comparisons based on morphometrics of the population with previous studies of the females and the first-stage juveniles of X. hunaniense with X. radicicola are given, and morphological variation in X. hunaniense populations are discussed. A revised polytomous key code of Loof and Luc (1990) for X. hunaniense identification is provided, i.e., A1- B4- C4- D4/5- E1- F2(3)- G2- H2-I3- J4- K2- L1. In addition, the sequence of the D2 and D3 expansion region of the 28S rRNA gene was analyzed and compared with sequences of closely related species downloaded from the NCBI database. Cluster analysis of sequences confirmed and supported the species identifications.     

Systematics and gene geography of Juniperus communis L. inferred from isoenzyme data

Using isoenzyme analysis, 35 populations of Juniperus communis L. from various parts of the Russian species range and by one population from Sweden and Alaska were studied. The total sample size was 1200 plants. As a result, the existence of J. communis var. oblonga in North Caucasus and J. communis var. depressa in North America was confirmed, but genetic differences between J. communis var. communis and J. communis var. saxatilis were not detected in the main part of the Russian species range (European part of Russia, Ural, Siberia). These populations proved to be genetically uniform with the same predominant allelic frequencies, which may evidence recent settling of this species from one of Central or East European refugium. J. communis var. saxatilis from northeastern Russia inhabiting the region behind Verkhoyansk mountain and Russian Far East showed considerable differentiation in frequencies of alleles at three loci and geographical subdivision. These populations also exhibit high intrapopulation variation. This can be connected with the refugium in this territory. The origin of this group is probably connected with migrations from Central Asia (Tibet) in the direction to northeastern Russia along mountains connecting Central and North Asia. It is also assumed that migrations of this species previously proceeded across the Beringian land bridge.     

Didymella pisi sp. nov., the teleomorph of Ascochyta pisi

The anamorphic pycnidial fungus Ascochyta pisi is one member of a species complex that causes Ascochyta blight of pea, a potentially devastating disease. The teleomorphic state of this fungus was induced under laboratory conditions. Using morphological and molecular characters, we placed the teleomorph within the genus Didymella as D. pisi and describe a heterothallic mating system using a PCR-based mating type assay and in vitro crosses. We compare D. pisi with other Didymella spp. with which it might be confused.     

Extent of Genetic Lesions of the Arginine and Pyrimidine Biosynthetic Pathways in Lactobacillus plantarum, L. paraplantarum, L. pentosus, and L. casei: Prevalence of CO2-Dependent Auxotrophs and Characterization of Deficient arg Genes in L. plantarum

Lactic acid bacteria require rich media since, due to mutations in their biosynthetic genes, they are unable to synthesize numerous amino acids and nucleobases. Arginine biosynthesis and pyrimidine biosynthesis have a common intermediate, carbamoyl phosphate (CP), whose synthesis requires CO₂. We investigated the extent of genetic lesions in both the arginine biosynthesis and pyrimidine biosynthesis pathways in a collection of lactobacilli, including 150 strains of Lactobacillus plantarum, 32 strains of L. pentosus, 15 strains of L. paraplantarum, and 10 strains of L. casei. The distribution of prototroph and auxotroph phenotypes varied between species. All L. casei strains, no L. paraplantarum strains, two L. pentosus strains, and seven L. plantarum strains required arginine for growth. Arginine auxotrophs were more frequently found in L. plantarum isolated from milk products than in L. plantarum isolated from fermented plant products or humans; association with dairy products might favor arginine auxotrophy. In L. plantarum the argCJBDF genes were functional in most strains, and when they were inactive, only one gene was mutated in more than one-half of the arginine auxotrophs. Random mutation may have generated these auxotrophs since different arg genes were inactivated (there were single point mutations in three auxotrophs and nonrevertible genetic lesions in four auxotrophs). These data support the hypothesis that lactic acid bacteria evolve by progressively loosing unnecessary genes upon adaptation to specific habitats, with genome evolution towards cumulative DNA degeneration. Although auxotrophy for only uracil was found in one L. pentosus strain, a high CO₂ requirement (HCR) for arginine and pyrimidine was common; it was found in 74 of 207 Lactobacillus strains tested. These HCR auxotrophs may have had their CP cellular pool-related genes altered or deregulated.     

LECANORA COMPALLENS AND L. SINUOSA,TWO NEW OVERLOOKED CORTICOLOUS LICHEN SPECIES FROM WESTERN EUROPE

Lecanora compallens and L. sinuosa, two corticolous lichens, are described as species new to science on the basis of numerous collections. Lecanora compallens is a sorediate species, known only as sterile. It is probably common in Western Europe, but overlooked because it is very similar to L. expallens. Chemically, the new species is identical with L. strobilina, of which it may represent a sorediate counterpart. Lecanora sinuosa is a new species similar to L. chlarotera and so far only known from a limited area in the Netherlands and Germany, although it may be overlooked elsewhere. Lecanora sinuosa is very close to L. hybocarpa, with which it shares thepulicaris -type epihymenium inspersed with fine crystals but differs markedly by its thick thallus and thick and sinuous apothecium margin.     

Leydigia louisi louisi Jenkin, 1934 in the Neotropics, L. louisi mexicana n.subsp. in the Central Mexican highlands

Leydigia louisi Jenkin, 1934 was recently redescribed by Kotov (2003). We found in Guatemala and South of Mexico populations of this species, similar with the typical African form. A re-examination of Daday's (1902) specimens from Patagonia demonstrated that his material, determined as L. leydigi, was also L. louisi. At the same time, we describe a new subspecies Leydigia louisi mexicana n.subsp. from four water bodies on the central Mexican plateau (State of Mexico). Parthenogenetic females, ephippial females, juvenile and adult males are described. SEM illustrations are given for L. louisi mexicana n.sp. It differs from the nominotypical subspecies by (1) ocellus of irregular shape; (2) chitinous insertions within distal segments of swimming setae far from bases of segments; (3) on IDL, 3 bunches of very long setules; (4) setules on distal lobe of limb II long; (5) longest seta of exopodite III with long, fine setules; (6) seta 1 of exopodite IV relatively short; (7) a two-lobed body on inner margin of limb V. Features 2–6 were never used in the genus previously, but were applied in the systematics of other genera of Chydoridae. The ranges of these two subspecies are, probably, the Neotropical and Holarctic regions.     

Trap Induction and Trapping in Eight Nematode-trapping Fungi (Orbiliaceae) as Affected by Juvenile Stage of <Emphasis Type="Italic">Caenorhabditis elegans</Emphasis>

This study measured trap induction and trapping on agar disks as affected by juvenile stages (J1, J2, J3, and J4) of the nematode Caenorhabditis elegans and by species of nematode-trapping fungi. Eight species of nematode-trapping fungi belonging to the family Orbiliaceae and producing four kinds of traps were studied: adhesive network-forming Arthrobotrys oligospora, A. vermicola, and A. eudermata, constricting ring-forming Drechslerella brochopaga, and Dr. stenobrocha, adhesive column-forming Dactylellina cionopaga, and adhesive knob-forming Da. ellipsospora, and Da. drechsleri. The number of traps induced generally increased with increasing juvenile stages of C. elegans. The ability to capture the juveniles tended to be similar among isolates that produced the same kind of trap but differed among species that produced different kinds of traps. Trapping by Dr. stenobrocha and Da. cionopaga was correlated with trap number and with juvenile stage. A. oligospora and A. vermicola respectively captured more than 92 and 88% of the J1, J3, and J4 but captured a lower percentage of J2. The knob-producing isolates captured more younger than elder juveniles. Partial correlation analyses demonstrated that the trap induction of the most fungal species positively correlated with the juvenile size and motility, which was juvenile stage dependent. Overall, trap induction and trapping correlated with C. elegans juvenile stage (size and motility) in six species of trapping fungi.     

Ecological status of the Mediterranean Juniperus phoenicea L. Relicts in the desert mountains of North Sinai,Egypt

Juniperus phoenicea L. is listed as threatened tree by IUCN Red List. In Egypt, J. phoenicea L. is the only conifer tree that is restricted to the three mountains of northern Sinai: Gabal El-Halal, Gabal El-Maghara and Gabal Yelleq. As a Mediterranean relict it has been included in a national list as target for conservation and management. To provide baseline information for the development of a conservation strategy, the present study aims at comparing the isolated populations of J. phoenicea and their associated plant composition and diversity at the three mountains. The application of TWINSPAN and DCA analysis techniques has resulted in identifying of four vegetation types associated with juniper, and each could be related to a specific geomorphologic habitat on a topographic gradient. Chiliadenus montanus and Zygophyllum dumosum characterized the slopes of smooth-faced rock outcrops in Wadi Abu Seyal (at 350–470 m altitude of Gabal El-Halal), Deverra tortuosa, Ephedra aphylla and Gymnocarpos decander inhabited together with the target species the soil pockets of north-facing slope in Neqeb Abu Hamam (at 600–700 m altitude of Gabal El-Halal), Stachys aegyptiaca and Moricandia nitens characterized the juniper occurrences in the runnels of Wadi Arar (at 450–560 m altitude of Gabal El-Maghara), and Artemisia herba-alba, Atriplex halimus and Reaumuria hirtella represent the stands on slope runnels (at 900–960 m) of Gabal Yelleq. The two vegetation types recognized at Gabal El-Halal had, on average, the highest species diversity, juniper density and cover. Juniper shows generally poor conditions of vitality at higher elevation (600–960 m) with a higher proportions of old and recent dead trees, and with the predominance of male individuals, as compared with the populations of Gabal El-Maghara and Gabal Yelleq. In contrast, the juniper populations at lower elevation (350–470 m) of Gabal El-Halal proved to be in best condition with mostly living foliage and reproductive branches. The differences in rock types and elevation among the three mountains reflect serious limitation on recruitment of J. phoenicea due to moisture availability. The results of this study showed that J. phoenicea is an endangered species and its conservation in northern Sinai mountains is a priority. For a successful conservation of this community it is highly recommended to preserve in particular the suitable habitats at Gabal El-Halal, but also the other stands merit conservation measures.     

Description of Thecavermiculatus carolynae n. sp. (Nematoda: Ataloderinae)

Thecavermiculatus carolynae n. sp. from the Pacific coastal area of California is described from female, male, and juvenile (J2) specimens reared on Festuca megalura Nutt. in the greenhouse. Differences between females of T. carolynae and those of its closest relative, T. gracililancea Robbins, 1978, include shorter stylet, shorter length, and lesser length of the vulval slit. Of these two species, males have been found only for T. carolynae. The habitats of the two species are vastly different: T. gracililancea has been found only in shallow clay soil near hilltops in an area subject to high summer temperatures where the soil becomes brick-like during the summer; T. carolynae has been found only in deep sandy soil of a creek bottom subject to cool summer temperatures where the soil remains moist.     

Phylogenetic Re-Evaluation of the Laurencia Complex (Rhodophyta) with a Description of L. Succulenta Sp. Nov. from Korea

Laurencia succulenta sp. nov. (Rhodophyta) is described from Korea. This species exhibits vegetative and reproductive structures typical of the genus, but is distinct from similar species in its epiphytic habitat and the fleshy, robust, thick and subcompressed thalli with basically distichous branching. In addition, it is readily distinguished from the most similar species, such as L. nipponica Yamada and L. okamurae Yamada, by the cystocarps with a somewhat protuberant ostiole. In a phylogenetic analysis of 47 species of the Laurencia Lamouroux complex from various localities around the world based on 49 morphological characters, four major clades (Laurencia, Chondrophycus palisadus (Yamada) Nam group, C. cartilagineus (Yamada) Garbary et Harper group and Osmundea Stackhouse assemblage), each of which forms a monophyletic group, were recognized. Among these, the Laurencia clade is basal to the overall assemblage, and is defined by the vegetative axis with four rather than two pericentral cells. The Osmundea clade is supported by autapomorphic characters for the genus, features associated with spermatangial formation of the filament type and tetrasporangial production from epidermal cells. By contrast, Chondrophycus, a genus characterized by a combination of features (vegetative axis with two pericentral cells, trichoblast-type spermatangial development and tetrasporangial production from pericentral cells), is paraphyletic, and the species were separated into two well-supported clades, the C. palisadus group and C. cartilagineus group. These clades are distinguished from each other by the position of the first pericentral cell relative to the trichoblast, the presence or absence of fertility at the second pericentral cells and number of sterile pericentral cells in the tetrasporangial axis, the pattern of formation of spermatangial branches on trichoblasts, post-fertilization feature associated with the formation time of the auxiliary cell, and, probably, the number of pericentral cells in the procarp-bearing segment. Of these features, the side position of the first pericentral cell in the latter group (a synapomorphy for the C. cartilagineus group plus Osmundea) suggests that the C. cartilagineus group is more closely related to Osmundea than to the C. palisadus group. This cladistic analysis indicates that Chondrophycus is not monophyletic, suggesting that the C. palisadus group should be separated from Chondrophycus at the genus level. Based on this result, Palisada (Yamada) stat. nov. is proposed for the group, together with an emendation of the generic delineation of Chondrophycus, and relevant nomenclatural changes for several Chondrophycus species are also included. In addition, Corynecladia J. Agardh is reinstated for the type species L. clavata Sonder.