Plant-Parasitic Nematodes of South Viet Nam

Between 1974 and 1978, 2,842 identifications of plant-parasitic nematodes were made from more than 1,700 soil and plant samples collected in eight provinces of South Viet Nam. Species in nine genera—Helicotylenchus, Criconemoides, Meloidogyne, Pratylenchus, Tylenchorhynchus, Hoplolaimus, Hirschmanniella, Xiphinema, and Rotylenchulus—comprised 96.1% of the identifications; the remaining 3.9% were species of 11 genera. Fourteen genera were associated with rice which was grown on about 2,500,000 ha in 1970. Of these, Ditylenchus, Hirschmanniella, and Meloidogyne were most important. Ditylenchus angustus caused severe damage to about 50,000 ha of flooded rice in the Mekong Delta in 1976. Hirschmanniella spp. were found in all samples examined from flooded rice fields. Meloidogyne spp. were common in rice seedbeds, upland rice, and rice not kept flooded continuously. Meloidogyne and Pratylenchus spp. were found in roots of 22 of the 32 crop plants sampled. Little or no attempt was made in South Viet Nam to control nematodes.     

Molecular phylogeny of Phoma and allied anamorph genera: Towards a reclassification of the Phoma complex

The present generic concept of Phoma is broadly defined, with nine sections being recognised based on morphological characters. Teleomorph states of Phoma have been described in the genera Didymella, Leptosphaeria, Pleospora and Mycosphaerella, indicating that Phoma anamorphs represent a polyphyletic group. In an attempt to delineate generic boundaries, representative strains of the various Phoma sections and allied coelomycetous genera were included for study. Sequence data of the 18S nrDNA (SSU) and the 28S nrDNA (LSU) regions of 18 Phoma strains included were compared with those of representative strains of 39 allied anamorph genera, including Ascochyta, Coniothyrium, Deuterophoma, Microsphaeropsis, Pleurophoma, Pyrenochaeta, and 11 teleomorph genera. The type species of the Phoma sections Phoma, Phyllostictoides, Sclerophomella, Macrospora and Peyronellaea grouped in a subclade in the Pleosporales with the type species of Ascochyta and Microsphaeropsis. The new family Didymellaceae is proposed to accommodate these Phoma sections and related anamorph genera. The present study demonstrated that Phoma radicina, the type species of Phoma sect. Paraphoma and Phoma heteromorphospora, the type species of Phoma sect. Heterospora can be assigned to the Phaeosphaeriaceae and Leptosphaeriaceae respectively.     

Occurrence,Parasitism, and Pathogenicity of Nematodes Associated with Sycamore (Platanus occidentalis L.)

Ten species of stylet-bearing nematodes were recovered in a survey of sycamore (Platanus occidentalis L. ) stands in Georgia. Helicotylenchus, Xiphinema, and Criconemoides were the genera found most frequently. Populations of Hoplolaimus galeatus, Scutellonema brachyurum, Helicotylenchus dihystera and H. pseudorobustus increased on greenhouse-grown sycamore, but Trichodorus christiei, Xiphinema americanum, Meloidogyne hapla, M. arenaria and M. incognita did not. Hoplolaimus galeatus and S. brachyurum are semi-endoparasites; H. dihystera and H. pseudorobustus are migratory endoparasites. Hoplolaimus galeatus caused extensive root necrosis and marked decrease of fresh weights of seedling roots and tops. Helicotylenchus dihystera and S. brachyurum produced only qualitatively different sparse and unhealthy root growth. Helicotylenchus pseudorobustus caused only a reduction in root surface area.     

Phylogenetic evaluation of Geomyces and allies reveals no close relatives of Pseudogymnoascus destructans, comb. nov., in bat hibernacula of eastern North America

White-nose syndrome (WNS)  of bats, caused by the fungus previously known as Geomyces destructans, has decimated populations of insectivorous bats in eastern North America. Recent work on fungi associated with bat hibernacula uncovered a large number of species of Geomyces and allies, far exceeding the number of described species. Communication about these species has been hindered by the lack of a modern taxonomic evaluation, and a phylogenetic framework of the group is needed to understand the origin of G. destructans and to target closely related species and their genomes for the purposes of understanding mechanisms of pathogenicity. We addressed these issues by generating DNA sequence data for the internal transcribed spacer (ITS) region, nuclear large subunit (LSU) rDNA, MCM7, RPB2, and TEF1 from a diverse array of Geomyces and allies that included isolates recovered from bat hibernacula as well as those that represent important type species. Phylogenetic analyses indicate Geomyces and allies should be classified in the family Pseudeurotiaceae, and the genera Geomyces, Gymnostellatospora, and Pseudogymnoascus should be recognized as distinct. True Geomyces are restricted to a basal lineage based on phylogenetic placement of the type species, Geomyces auratus. Thus, G. destructans is placed in genus Pseudogymnoascus. The closest relatives of Pseudogymnoascus destructans are members of the Pseudogymnoascus roseus species complex, however, the isolated and long branch of P. destructans indicates that none of the species included in this study are closely related, thus providing further support to the hypothesis that this pathogen is non-native and invasive in eastern North America. Several conidia-producing isolates from bat hibernacula previously identified as members of Pseudeurotium are determined to belong to the genus Leuconeurospora, which is widespread, especially in colder regions. Teberdinia hygrophila is transferred to Pseudeurotium as Pseudeurotium hygrophilum, comb. nov., in accordance with the one name per fungus system of classification, and two additional combinations are made in Pseudogymnoascus including Pseudogymnoascus carnis and Pseudogymnoascus pannorum. Additional sampling from other regions of the world is needed to better understand the evolution and biogeography of this important and diverse group of fungi.     

Nematodes Attacking Cultivars of Peach in North Carolina

Criconemoides xenoplax and Meloidogyne incognita were the nematode species most frequently associated with peach in North Carolina. Other nematodes often found in high numbers on that crop were Pratylenehus vulnus, Helicotylenchus spp., Trichodorus christiei, Xiphinema amerieanum and Tylenchorhynchus claytoni. P. vulnus and P. penetrans reproduced well on rootstocks of 21 peach cultivars tested in the greenhouse. P. zeae, P. brachyurus, P. coffeae and P. scribneri decreased or increased only slightly in most instances. C. xenoplax increased as much as 330-fold and reproduced on all cultivars tested. In a field experiment with six peach cultivars and moderate numbers of P. brachyurus, P. vulnus, C. xenoplax, and M. incognita, only M. incognita caused significant stunting in 30 months. This nematode increased only on root-knot susceptible cultivars, whereas the other nematodes followed the same patterns observed in the greenhouse. In a second field experiment, seedlings were stunted significantly by high numbers of C. xenoplax during an 18-month period.     

Influence of host plants on feeding, growth and reproduction of Papilio polytes (The common mormon)

We studied the feeding, growth and reproductive behaviour of Papilio polytes (common mormon butterfly) on five different host plants, Murraya koenigii, Toddalia asiatica, Glycosmis pentaphylla, Aegle marmelos and Citrus medica. The growth rate of P. polytes was fastest on M. koenigii followed by T. asiatica, C. medica, G. pentaphylla and A. marmelos. We related this to the nutrient contexts of the five plants. The plants T. asiatica and C. medica had higher water contents, which influenced the growth rate of the insect. M. koenigii was found to contain rich quantities of carbohydrate. M. koenigii, T. asiatica and C. medica were also rich in protein when compared to A. marmelos and G. pentaphylla. Total amino acid levels were comparatively higher in M. koenigii, T. asiatica, C. medica rather than A. marmelos and G. pentaphylla.     

Ecological Study of Nematode Parasitism in Ips Beetles from California and Idaho

Nematodes found in Ips paraconfusus from ponderosa pine in California were an undescribed species of Parasitaphelenchus, Contortylenchus elongatus, C. reversus, and C. brevicomi. C. elongatus, the most commonly found contortylenchid, was present in 98.2% of the contortylenchid-parasitized beetles. Only one nematode parasite of the gut, a Parasitorhabditis sp., was isolated. Although significant differences in parasitism were observed, they were by collection sites, rather than by elevation or bole sources (slash or standing). Significant changes in parasitism between fall and spring collections were observed but not at every site. Nematode parasitism in the F₁ generation of I. paraconfusus by Parasitaphelenchus, Contortylenchus, or Parasitorhabditis increased or decreased from the parent generation depending upon the experiment.Nematode parasites from I. pini included an undescribed Parasitaphelenchus sp., two undescribed Contortylenchus spp., C. reversus and Parasitylenchus (= Neoparasitylenchus) ovarius from the hemocel, and Parasitorhabditis ipini from the gut. Parasitaphelenchus sp. was found in 99% and 45.3% of the beetles from Idaho and California, respectively. Of the 1,000 I. pini from Idaho and California, 157 were parasitized by the contortylenchid species or P. ovarius.     

Endophytic occupation of peanut root nodules by opportunistic Gammaproteobacteria

Several bacterial isolates were recovered from surface-sterilized root nodules of Arachis hypogaea L. (peanut) plants growing in soils from Córdoba, Argentina. The 16S rDNA sequences of seven fast-growing strains were obtained and the phylogenetic analysis showed that these isolates belonged to the Phylum Proteobacteria, Class Gammaproteobacteria, and included Pseudomonas spp., Enterobacter spp., and Klebsiella spp. After storage, these strains became unable to induce nodule formation in Arachis hypogaea L. plants, but they enhanced plant yield. When the isolates were co-inoculated with an infective Bradyrhizobium strain, they were even found colonizing pre-formed nodules. Analysis of symbiotic genes showed that the nifH gene was only detected for the Klebsiella-like isolates and the nodC gene could not be amplified by PCR or be detected by Southern blotting in any of the isolates. The results obtained support the idea that these isolates are opportunistic bacteria able to colonize nodules induced by rhizobia.     

Alkaloids in the venom of Messor ants

Tobacco alkaloids of the anabasine type have been found or confirmed in the venom of five species of arid-dwelling Messor ants. They are frequently accompanied by alkylpyrazines. Messor mediorubra contains four alkaloids, with anabasine the major component and also minor pyrazines. Anabasine was found alone in the venom of Messor semirufus and confirmed in Messor ebeninus. Messor rugosus from Tel Aviv contained a mixture of alkaloids and pyrazines, but those from Ein Yahav contained 3-ethyl-2,5-dimethylpyrazine, some 2-phenylethylamine, and N-ethylidene 2-phenylethylamine. Messor arenarius is confirmed as having a complex but variable mixture of alkaloids and pyrazines.     

Frequency of ace, epa and elrA Genes in Clinical and Environmental Strains of Enterococcus faecalis

Surface proteins play an important role in the pathogenesis of enterococcal infections. Some of them are candidates for a vaccine, e.g., the frequency of endocarditis in rats vaccinated with Ace protein was 75 % as 12 opposed to 100 % in those who weren’t. However, there are other components of enterococcal cells, such as Epa antigens or internalin-like proteins, which may be used in the prophylaxis of infections caused by them. However, also other virulence factors and resistance to antibiotics are important during enterococcal infection. Therefore, the relevance of ace, epa, elrA, other virulence genes, as well as resistance to antibiotics was investigated. 161 Enterococcus faecalis strains isolated from teaching hospitals in Lodz, cultured according to standard microbiological methods, were investigated for the presence of genes encoding surface proteins by PCR. Results were analyzed with χ² test. The elrA gene was found in all clinical and environmental strains, the ace gene was also widespread among E. faecalis (96.9 %). Both tested epa genes were found in the majority of isolates (83.25 %). There was correlation between the presence of esp and ace genes (p = 0.046) as well as between epa and agg genes (p = 0.0094; χ² test). The presence of the genes encoding surface proteins investigated in our study in the great majority of isolates implies that they would appear to be required during E. faecalis infection. Therefore, they could be excellent targets in therapy of enterococcal infections or, as some studies show, candidates for vaccines.     

Multilocus phylogenetic analysis of the genus Aeromonas

A broad multilocus phylogenetic analysis (MLPA) of the representative diversity of a genus offers the opportunity to incorporate concatenated inter-species phylogenies into bacterial systematics. Recent analyses based on single housekeeping genes have provided coherent phylogenies of Aeromonas. However, to date, a multi-gene phylogenetic analysis has never been tackled. In the present study, the intra- and inter-species phylogenetic relationships of 115 strains representing all Aeromonas species described to date were investigated by MLPA. The study included the independent analysis of seven single gene fragments (gyrB, rpoD, recA, dnaJ, gyrA, dnaX, and atpD), and the tree resulting from the concatenated 4705 bp sequence. The phylogenies obtained were consistent with each other, and clustering agreed with the Aeromonas taxonomy recognized to date. The highest clustering robustness was found for the concatenated tree (i.e. all Aeromonas species split into 100% bootstrap clusters). Both possible chronometric distortions and poor resolution encountered when using single-gene analysis were buffered in the concatenated MLPA tree. However, reliable phylogenetic species delineation required an MLPA including several “bona fide” strains representing all described species.     

Characterization of 10 MADS-box genes from Pyrus pyrifolia and their differential expression during fruit development and ripening

We cloned 10 Japanese pear (Pyrus pyrifolia) MIKC-type II MADS-box genes, and analyzed their expression during fruit development and ripening. PpMADS2-1 was APETALA (AP)1-like; PpMADS3-1 was FRUITFULL (FUL)/SQUAMOSA (SQUA)-like; PpMADS4-1 was AGAMOUS-like (AGL)6; PpMADS5-1 and PpMADS8-1 were SUPPRESSOR OF OVEREXPRESSION OF CONSTANS (SOC)-like; PpMADS9-1, PpMADS12-1, PpMADS14-1 and PpMADS16-1 were SEPALLATA (SEP)-like; while PpMADS15-1 was AGL/SHATTERPROOF (SHP)-like. Phylogenetic analysis showed their grouping into five major clades (and 10 sub-clades) that was consistent with their diverse functional types. Expression analysis in flower tissue revealed their distinct putative homeotic functional classes: A-class (PpMADS2-1, PpMADS3-1, PpMADS4-1, and PpMADS14-1), C-class (PpMADS15-1), E-class (PpMADS9-1, PpMADS12-1, and PpMADS16-1) and E (F)-class (PpMADS5-1 and PpMADS8-1). Differential gene expression was observed in different fruit tissues (skin, cortex and core) as well as in the cortex during the course of fruit development and ripening. Collectively, our results suggest their involvement in the diverse aspects of plant development including flower development and the course of fruit development and ripening.     

Interactions of Concomitant Species of Nematodes and Fusarium oxysporum f. sp. vasinfectum on Cotton

Meloidogyne incognita, Hoplolaintus galeatus, and North Carolina and Georgia populations of Belonolaimus longicaudatus were introduced singly and in various combinations with Fusarium oxysporum f. sp. vasinfectum on wilt-susceptible ''Rowden'' cotton. Of all the nematodes, the combination of the N. C. population of B. longicaudatus with Fusarium promoted greatest wilt development. H. galeatus had no effect on wilt. With Fusarium plus M. incognito or B. longicaudatus, high nematode levels promoted greater wilt than low levels. The combination of either population of B. longicaudatus with M. incognita and Fusarium induced greater wilt development than comparable inoculum densities of either nematode alone or where H. galeatus was substituted for either of these nematodes. Nematode reproduction was inversely related to wilt development. Without Fusarium, however, the high inoculum level resulted in greater reproduction of all nematode species on cotton. Combining M. incognita with B. longicaudatus or H. galeatus gave mutually depressive effects on final nematode populations. The interactions of H. gateatus with B. longicaudatus varied with two populations of the latter.     

Natural Occurrence of Entomogenous Nematodes in Tennessee Nursery Soils

To isolate potential insect biocontrol agents, entomogenous nematodes were surveyed in Tennessee plant nurseries in 1991. Soil samples from 113 nursery sites were baited with greater wax moth (Galleria mellonella) larvae, house cricket (Acheta domesticus) adults, lesser mealworm (Alphitobius diaperings) adults, and house fly (Musca domestica) larvae. Heterorhabditis bacteriophora and Steinernema carpocapsae were each recovered from 17 soil samples. Heterorhabditis bacteriophora was more common in habitats with crape myrtle (Lagerstroemia indica) and Chinese juniper (Juniperus chinensis) than other nursery plants, and S. carpocapsae was more frequently recovered from habitats with juniper and Southern magnolia (Magnolia grandiflora). Bulk density, electrical conductivity, organic matter, pH, temperature, and moisture content of the entomogenous-nematode positive soil samples were compared. Other nematode genera recovered with insect baits included Rhabditis sp., Pelodera sp., Cryptaphelenchoides sp., and Mesodiplogaster sp., which was recovered from a greater percentage of soil samples than the other five genera.     

Essential oil composition of Hypericum L. species from Southeastern Serbia and their chemotaxonomy

The essential oils of the aerial parts of nine species of Hypericum (Hypericum barbatum, Hypericum hirsutum, Hypericum linarioides, Hypericum maculatum, Hypericum olympicum, Hypericum perforatum, Hypericum richeri, Hypericum rumeliacum and Hypericum tetrapterum), collected from different locations in Southeast Serbia, were obtained by steam distillation and analyzed by GC and GC–MS. The essential oils investigated were characterized by a high content of non-terpene compounds and a low content of monoterpenes. The contents of non-terpenes, monoterpenes and sesquiterpenes in oils of the species H. barbatum, H. richeri and H. rumeliacum (section Drosocaprium) were similar and these oils were characterized by high contents of fatty acids. The oils of H. hirsutum and H. linarioides (section Taeniocarpium) contained a high percentage of n-nonane. There were similarities in contents of non-terpenes and sesquiterpenes in oils of species that belong to the section Hypericum (H. maculatum, H. perforatum and H. tetrapterum). The oil of H. olympicum differed from others by higher terpene content. A comparison was also carried out of the chemical composition of the essential oils from flower, leaf and stem of H. perforatum and it revealed that the highest concentration of non-terpene compounds was found in the flower and stem oil, while a high concentration of sesquiterpenes was characteristic for leaf oil. There were significant differences in the concentrations of the same compounds in the essential oils of H. maculatum, H. olympicum and H. perforatum, collected in different years from the same location which could be explained by seasonal differences. All data were statistically processed with principal component analysis and cluster analysis. The main conclusion from the above data is that genetic and environmental factors both play a role in determining the composition of essential oils of the Hypericum species studied.