Localized Influence of Heterodera glycines on Sudden Death Syndrome of Soybean

Half-root tests were established to examine the association between Heterodera glycines and the blue strain of Fusarium solani, the causal agent of sudden death syndrome (SDS) of soybean. Two independent root systems were established for soybean ''Coker 156'' and inoculated (half root/half root) with F. solani, H. glycines, both organisms on opposite root halves, both organisms on one root half, or neither one. Foliar symptoms were more severe for plants inoculated with both organisms on one root half than on opposite root halves or F. solani alone. Root necrosis ratings were more severe when both pathogens were combined on one root half than on opposite root halves. Heterodera glycines population development was reduced by the combination of both pathogens on one root half compared to opposite root halves or H. glycines alone, regardless of inoculation time.     

Hybridization betweenPsidium guajava andP. guineense (Myrtaceae)

Psidium guajava andP. guineense are characterized, illustrated and contrasted. Population samples ofPsidium guajava andP. guineense and apparent hybrids were collected from four localities: two in Mexico, one in Honduras, and one in Argentina. Seven morphological characters, one micromorphological (stomatal density), and one chemical character (presence or absence of myricetin) that distinguish the parent species were found to be useful in the evaluation of specimens and population variation. Results were consistent with the hypothesis that hybridization occurs between these species, but in three localities it may not go beyond Fl ’s. Back crossing between hybrids andP. guajava was not detected at any locality.     

Comparison of Populations of Pratylenchus brachyurus Based on Isozyme Phenotypes

Enzymes from females of five Pratylenchus brachyurus populations and one P. scribneri population were analyzed by isoelectric focusing electrophoresis. Of the 18 enzyme systems investigated, only malate dehydrogenase (MDH), phosphoglucomutase (PGM), and phosphoglucose isomerase (PGI) were detected from all five P. brachyurus populations and P. scribneri. Faint bands were detected for isocitrate dehydrogenase and phosphogluconate dehydrogenase from one P. brachyurus population. Three distinct phenotypic groups were found in the MDH and PGM systems for P. brachyurus populations, but only a single electromorph was detected for PGI. Multiple electromorphs for MDH, PGM, and PGI were detected for P. scribneri; there was no similarity among these patterns with those from P. brachyurus. No phenotypic differences in PGI were observed between females and mixed juveniles of one population of P. brachyurus.     

Phytoparasitic Nematodes Adjacent to Established Strawberry Plantations

Plant-nematode populations associated with uncultivated vegetation, adjacent strawberry plants, and alternate crop sites were studied at three locations in Minnesota. At one site (Forest Lake), Paratylenchus projectus, Meloidogyne hapla, and Pratylenchus tenuis were frequently associated with the roots of native vegetation. These nematode species were also present in adjacent strawberry beds. Among alternate crops observed, oats and muskmelon usually supported the fewest nematodes although moderate densities of Xiphinema americanum and P. tenuis were found at one location in plots planted to oats. Pratylenchus tenuis was also found on rye at one location.     

Genotyping of Clostridium perfringens isolated from calves with neonatal diarrhea

Neonatal diarrhea is one of the main causes of losses in cattle herds. Clostridium perfringens is a widespread enteropathogen, and is responsible for many animal diseases such as bovine neonatal diarrhea. Fecal samples from 141 diarrheic calves and 129 healthy calves, aged up to 28 days and belonging to three herds were examined. Rates of culture positivity were 36.2% and 30.2% for diarrheic and nondiarrheic calves, respectively. Multiple isolates from primary isolation plates were subjected to simultaneous genotyping by multiplex PCR, with primers amplifying fragments of alpha (cpa), beta (cpb), epsilon (etx), iota (itxA), enterotoxin (cpe) and beta2 (cpb2) toxin-encoding genes. Only 17/51 (33.3%) and 17/39 (43.6%) of these mixtures from diarrheic and nondiarrheic calves, respectively, yielded genotype information, suggesting that this may not be a viable approach to genotyping of isolates. Fourteen isolate mixtures from animals with diarrhea had only cpa (type A), one had cpa and cpb2 (type A beta2 positive), one with cpa, itxA, and cpb2 (type E, beta2 positive), and one with cpa, etx, itxA, and cpb2 toxin producing strains. Among 17 isolate mixtures from healthy calves, 10 were exclusively type A, one was type A cpb2 positive, two were type E, three were type E cpb2 positive, and one was types D and E cpb2 positive. There was no correlation between isolation of a given toxin type and the presence of diarrhea.     

Flavonoids of Derris obtusa: Aurones and auronols

Heptacosanol and sitosterol, one chalcone, one flavone, one 3-O-methyl-flavonol, four aurones and two 3-O-methylauronols were isolated from the root bark of Derris obtusa. The latter compounds, called Derriobtusones A and B, are the first auronols found in nature. Structures were established through chemical and spectral means. Mass spectral fragmentation schemes are suggested for aurones and auronols.     

Differential Response to Root-Knot Nematodes in Prunus Species and Correlative Genetic Implications

Responses of 17 Prunus rootstocks or accessions (11 from the subgenus Amygdalus and 6 from the subgenus Prunophora) were evaluated against 11 isolates of Meloidogyne spp. including one M. arenaria, four M. incognita, four M. javanica, one M. hispanica, and an unclassified population from Florida. Characterization of plant response to root-knot nematodes was based on a gall index rating. Numbers of females and juveniles plus eggs in the roots were determined for 10 of the rootstocks evaluated against one M. arenaria, one M. incognita, one M. javanica, and the Florida isolate. These 10 rootstocks plus Nemaguard and Nemared were retested by growing three different rootstock genotypes together in containers of soil infested individually with each of the above four isolates. Garfi and Garrigues almonds, GF.305 and Rutgers Red Leaf peaches, and the peach-almond GF.677 were susceptible to all isolates. Differences in resistance were detected among the other rootstocks of the subgenus Amygdalus. The peach-almond GF.557 and Summergrand peach were resistant to M. arenaria and M. incognita but susceptible to M. javanica and the Florida isolate. Nemaguard, Nemared, and its two hybrids G x N no. 15 and G x N no. 22 were resistant to all but the Florida isolate. In the subgenus Prunophora, Myrobalan plums P.1079, P.2175, P.2980, and P.2984; Marianna plum 29C; and P. insititia plum AD.101 were resistant to all isolates. Thus, two different genetic systems of RKN resistance were found in the subgenus Amygdalus: one system acting against M. arenaria and M. incognita, and another system also acting against M. javanica. Prunophora rootstocks bear a complete genetic system for resistance also acting against the Florida isolate. The hypotheses on the relationships between these systems and the corresponding putative genes of resistance are presented.     

Immunological properties of entomocidal protein of Bacillus thuringiensis and its insecticidal activity

The immunological properties of the proteinaceous component of the parasporal crystal (δ-endotoxin) of Bacillus thuringiensis var. kurstaki were analyzed by rocket immunoelectrophoresis. Two antisera, one against the k-l-type crystal containing two components, and the other against the k-73-type crystal containing one component, were made in rabbits. The antigens consisting of purified and dissociated crystals were run in electrophoresis with these two antisera. The ratio between the two peak heights of precipitin lines, which were formed by the dissociated crystal of one B. thuringiensis isolate in two antisera, was compared with the ratios of other isolates under identical conditions. The difference in the ratio reflected a difference in the structure of the crystal component and correlated closely with the insecticidal activity spectrum. This method can be used to evaluate a newly isolated B. thuringiensis, and it can further differentiate the isolates which have been classified as one serotype.     

Aerosols as a Source of Widespread Mycoplasma Contamination of Tissue Cultures

Mycoplasma isolates were cultured from 15 antibiotic-free cell cultures obtained from a single laboratory. Complement-fixation tests showed that these isolates were antigenically related to each other but were unrelated to M. hominis type 1, M. hominis type 2, M. arthritidis, M. laidlawii type B, Mycoplasma sp. H.Ep. #2 (Barile), or M. salivarium. Examination of serum used to feed the infected cell lines revealed no Mycoplasma. Infection resulting from cross-contamination by a single Mycoplasma strain from one cell culture to another was investigated. Although the organisms were not found in the air over the work area, aerosols containing these contaminants were produced in tissue culture bottles during the trypsinization of cell monolayers. The minimal infectious dose of Mycoplasma for tissue cultures was measured, and it was determined that one organism was capable of initiating an infection in a tissue culture. The pattern of contamination and the small dose required for infection indicated that Mycoplasma contamination was spread from one tissue culture to another via aerosols. It was demonstrated that Mycoplasma can be transferred from one cell culture to another through the use of a common burette for dispensing medium.     

Serological Diversity Within a Terrestrial Ammonia-Oxidizing Population

Fluorescent antibodies (FAs) prepared against 16 ammonia-oxidizing nitrifying bacteria were examined as to cross-reactivity in heterologous FA staining tests. Virtually all cross-reactions to the seven Nitrosomonas FAs were confined to the Nitrosomonas isolates. The five Nitrosospira, three Nitrosolobus, and one Nitrosovibrio FAs stained isolates only within their respective genera. Within each genus shared antigens were more common among Nitrosomonas and Nitrosolobus than among Nitrosospira isolates. Isolates obtained from a single soil were stained to examine the effectiveness of the suite of FAs for study of a given ammonia-oxidizing population. Of the seven Nitrosomonas isolates, five were stained effectively through the use of four FAs; six of twelve Nitrosospira isolates were identifiable through use of four different FAs. The one Nitrosolobus isolate was stained well only by one (its homologous) FA.     

Microbiological spoilage of mayonnaise and salad dressings

Saccharomyces bailii was isolated from two-thirds of the spoiled mayonnaise and salad dressing samples examined. Most of the rest were spoiled by Lactobacillus fructivorans. However, one sample contained large numbers of both S. bailii and L. plantarum. Two of the spoiled samples also contained small numbers of bacilli. Bacillus subtilis, B. pumilis, B. polymyxa, B. megaterium, and B. licheniformis were found in one sample and B. subtilis and B. pumilis in another. Small numbers of B. subtilis and B. licheniformis were also present in one unspoiled sample. Several media were evaluated for the isolation of L. fructivorans. S. bailii and L. fructivorans vigorously fermented glucose. The concentration of glucose in the spoiled samples ranged from 0 to 38.5 g/kg and from 1.3 to 17.8 g/kg for the unspoiled samples.     

A chemosystematic study of the phenolics of Sonchus

Thirty-three Sonchus, one Embergeria, one Babcockia and five Taeckholmia species were surveyed for their phenolic constituents. The coumarins scopoletin and aesculetin were found as major constituents of Embergeria, Babcockia and Taeckholmia species, and in lesser amount in some Sonchus species. Six flavone glycosides were identified: apigenin 7-glucuronide, apigenin 7-rutinoside, luteolin 7-glucoside, luteolin 7-glucuronide, luteolin 7-rutinoside and luteolin 7-glucosylglucuronide and the systematic significance of their distribution is discussed.     

In vitro efficacies of oils, silicas and plant preparations against the poultry red mite Dermanyssus gallinae

The aim of this study was to test the effectiveness of physically acting substances (oils and silicas) and plant preparations for the control of the poultry red mite Dermanyssus gallinae (De Geer 1778). Reproduction and survival of fed D. gallinae females were evaluated in vitro for a total of 168 h using the “area under the survival curve” (AUC) to compare survival of the mites between treatments. Four oils (two plant oils, one petroleum spray oil and diesel), one soap, three silicas (one synthetic amorphous silica, one diatomaceous earth (DE) and one DE with 2% pyrethrum extract) and seven plant preparations (derived from Chrysanthemum cineariaefolium, Allium sativum, Tanacetum vulgare, Yucca schidigera, Quillaja saponaria, Dryopteris filix-mas, and Thuja occidentalis) were tested at various concentrations. All the oils, diesel and soap significantly reduced D. gallinae survival. All silicas tested inhibited reproduction. DE significantly reduced mite survival, but amorphous silica was less effective in vitro. Except for pure A. sativum juice and the highest concentration of C. cineariaefolium extract, the plant preparations tested resulted in statistically insignificant control of D. gallinae.     

Cytogenetics of Amaryllidaceae species: heterochromatin evolution in different ploidy levels

Species belonging to the Amaryllidaceae (Zephyranthes and Habranthus) were analyzed by banding with chromomycin A3 (CMA)/4,6-diamidino-2-phenylindole (DAPI) fluorochromes. The patterns of bands were studied in seven species of Zephyranthes Herb. and one of Habranthus Herb. Subterminal and interstitial DAPI+ bands were observed in Z. robusta 2n?=?12 and Z. brachyandra 2n?=?24. Other species showed no AT-rich heterochromatin. In species with 2n?=?12, CMA+ bands were observed on one chromosome pair of Z. robusta and Zephyranthes sp., while in Z. sylvatica an additional small terminal band in the fifth chromosome pair was observed. Z. rosea and Z. grandiflora presented with 2n?=?24 and had four CMA+ bands, while in Z. brachyandra, with 2n?=?24?+?1B, there were eight interstitial dot bands and a larger terminal band in the short arm of the B chromosome. Z. candida with 2n?=?38 presented CMA+ heterochromatin blocks on the long arms of five metacentric pairs and in the short arm of one of the submetacentric pairs; in addition a terminal band was observed on the long arm of one of the homologues of a larger submetacentric pair. H. itaobinus showed a heterozygous pair revealing a strong CMA+ band in only one of the homologues, likely a nucleolus organizing region. Taxonomic implications and karyotype evolution of this group are discussed and correlated with previous data from the literature.     

Isolation and Characterization of Methicillin-Resistant Staphylococcus aureus Strains from Louisiana Retail Meats

We investigated the prevalence of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in 120 retail meat samples from 30 grocery stores in Baton Rouge, LA. S. aureus strains were recovered from 45.6% of pork samples and 20% of beef samples, whereas MRSA strains were isolated from six meat samples (five pork samples and one beef sample). The MRSA isolates were of two strain types (clones), one harboring Panton-Valentine leucocidin and belonging to pulsed-field gel electrophoresis type USA300 and the other one belonging to USA100.     

Phytochemical and chemotaxonomic investigation from the roots of Anemone vitifolia Buch.-Ham. (Ranunculaceae)

Phytochemical investigation from the roots of Anemone vitifolia Buch.-Ham. led to the isolation of eight compounds, including one triterpenoid saponin, two sugars, one coumarin, one amide, one saturated alkane, one olefin and one fatty acid. The structures of these metabolites were elucidated by spectroscopic data and comparisons with the data available in the literature. Among them, compound 7 ((6Z,9Z,12Z)-6,9,12-Eicosatriene) was isolated for the first time as a natural product. Furthermore, compounds 2 (D-(+)-raffinose), 3 (mixture of β-D-fructopyranose and β-D-fructofuranose) and 5 (bonaroside) were obtained from the Ranunculaceae family for the first time. Compounds 4 (siderin) and 6 (n-hexadecane) were isolated from A. vitifolia for the first time. The chemotaxonomic significance of the isolated compounds was discussed.     

Isolation of Fungi from Heterodera glycines and in vitro Bioassays for Their Antagonism to Eggs

Twenty fungi were assayed in vitro for antagonism to eggs of Heterodera glycines. Eight of the fungi were isolated from cysts or eggs of H. glycines during the current study, one was isolated from Panagrellus redivivus, and eleven were obtained from other researchers or collections. The bioassays were conducted on eggs from nematodes that had been grown monoxenically on excised root tips. Phoma chrysanthemicola, one strain of Verticillium chlamydosporium, and one strain of V. lecanii caused a decrease (P < 0.01, P < 0.05, P < 0.05, respectively) in the number of viable eggs, although no hyphae were observed colonizing live eggs. Trichoderma polysporum infected live eggs but enhanced (P < 0.05) egg survival. Acremonium bacillisporum, Chaetomium sp., Drechmeria coniospora (two strains), Epicoccum sp., Exophiala jeanselmei, Fusarium sp., Neocosmospora vasinfecta, Scytalidium fulvum, Trichoderma harzianum (two strains), V. chlamydosporium (one strain), V. lecanii (three strains), and an unidentified fungus did not measurably affect egg viability, even though hyphae of five of these fungi were seen in live eggs. The bioassay provides a useful step in the selection of a biological control agent for this major nematode pest.     

Bacterial community on ectomycorrhizal roots of Laccaria laccata in a chestnut plantation

This study investigated the bacterial communities present on the ectomycorrhizal roots of Laccaria laccata using culture- and cloning-based methods. One soil core was collected from five locations where sporocarps of L. laccata occurred in a chestnut plantation. A total of 10 and 50 ectomycorrhizal root tips of L. laccata were collected from each soil core and subjected to bacterial isolation and cloning. In total, 223 isolates were classified into 35 molecular operational taxonomic units (MOTUs). The majority of MOTUs were infrequent, whereas one MOTU each of Rhizobium and Bradyrhizobium were common across the sampling locations. In the cloning-based method, 77 MOTUs were detected. The majority of these MOTUs were infrequent, whereas one MOTU each of Bradyrhizobium and one taxon of Gammaproteobacteria was common across the sampling locations. These results indicate that Bradyrhizobium is a common member of bacteria present on the ectomycorrhizal roots of L. laccata in the chestnut plantation.     

Occurrence of ant (hymenoptera, formicidae) and aphid (homoptera, aphidinea) syninclusions in Saxonian and Rovno ambers

Ant species found as syninclusions with aphids in Rovno and Saxonian ambers are listed for the first time. In a collection of 143 pieces of Rovno amber with worker ants, aphids were found in only one syninclusion with Ctenobethylus goepperti (Mayr). In a collection of Saxonian amber, 53 aphids of the genus Germaraphis were found in 152 amber pieces with ant workers; five syninclusions with aphids contained 19 specimens of C. goepperti; two syninclusions contained ten specimens of Lasius schiefferdeckeri Mayr; and three syninclusions contained one specimen each: one Camponotus mengei Mayr, one Anonichomyrma constricta (Mayr), and one Formica flori Mayr. Considering that the Rovno and Saxonian collections are nearly equal in the number of amber pieces, the fundamental difference in the number of syninclusions is striking. The number of C. goepperti in syninclusions is disproportionately high; the proportion of syninclusions with aphids and C. goepperti is 1.6 times that of all inclusions of workers of this species. They are found in 31% of examined inclusions of workers in the collection of the Saxonian amber, but in 50% of syninclusions of the entire collection. This strongly suggests that C. goepperti and Germaraphis formed an association during their lifetime.