N,N′-bis-(2,3-Methylenedioxyphenyl)urea and N,N′-bis-(3,4-methylenedioxyphenyl)urea enhance adventitious rooting in Pinus radiata and affect expression of genes induced during adventitious rooting in the presence of exogenous auxin

We have analyzed the effect of N,N′-bis-(2,3-methylenedioxyphenyl)urea (2,3-MDPU) and N,N′-bis-(3,4-methylenedioxyphenyl)urea (3,4-MDPU), two symmetrically substituted diphenylurea derivatives with no auxin or cytokinin-like activity, on the rooting capacity of Pinus radiata stem cuttings. Results indicate that both diphenylurea derivatives enhance adventitious rooting in the presence of exogenous auxin (indole-3-butyric acid, IBA), even at low auxin concentration, in rooting-competent cuttings, but have no effect on the adventitious rooting of low or null competent-to-root cuttings. Histological analyses show that, in the simultaneous presence of MDPUs and low concentration of exogenous auxin, adventitious root formation is induced in the cell types that retain intrinsic competence to form adventitious roots in response to auxin. The time course of cellular events leading to root formation and the time of root emergence are closely similar to that observed in cuttings treated only with higher auxin concentration. In addition, the mRNA level of a P. radiata SCARECROW-LIKE gene, which is significantly induced in the presence of the optimal concentration (10 μM) of exogenous auxin needed for cuttings to root, is increased in the presence of MDPUs and low concentration of exogenous auxin (1 μM). The expression of a P. radiata SHORT-ROOT gene in rooting-competent cuttings during adventitious rooting is also affected by the presence of MDPUs when combined with auxin. As MDPUs do not affect the expression of either gene in the absence of exogenous auxin, but only in its presence, we suggest that MDPUs could interact, directly or indirectly, with the auxin-signalling pathways in rooting-competent cuttings during adventitious rooting.     

Phylogeographic patterns in Leccinum sect. Scabra and the status of the arctic-alpine species L. rotundifoliae

We investigated inter- and intraspecific phylogenetic relationships in the ectomycorrhizal fungal genus Leccinum section Scabra. Species of this section are exclusively associated with Betula and occur throughout the Northern Hemisphere. We compared the phylogenetic relationships of arctic, alpine, boreal and temperate accessions of section Scabra based on DNA sequences of the single-copy nuclear gene Gapdh and the multiple-copy nuclear region 5.8S-ITS2. Exclusively arctic lineages were not detected in species that occur both in arctic-alpine or boreal regions, except in L. rotundifoliae that was restricted to cold climates. L. scabrum and L. holopus showed an intercontinental phylogeographic pattern, and L. variicolor showed a pattern unrelated to geographical distribution. Molecular clock estimates indicated that L. rotundifoliae is as old as other species in section Scabra. Individual gene trees suggest that interspecific hybridisation occurred several times in the evolution of section Scabra.     

Supplemental risk evaluations of Puccinia jaceae var. solstitialis for biological control of yellow starthistle

Additional tests of native North American Cirsium species, Saussurea americana, and modern safflower cultivars (Carthamus tinctorius) were requested by regulators and specific interest groups during the risk assessment of foreign isolates of Puccinia jaceae var. solstitialis for biological control of yellow starthistle (YST, Centaurea solstitialis) in the United States. These tests supplement an earlier, extensive host range determination that established P. jaceae from YST as generally host specific and potentially useful for biological control. The additional research was in response to potential hazards identified in an earlier study, changes in safflower cultivars, and concern that P. jaceae might cause a safflower seedling disease similar to hypocotyl infections from infestation by Puccinia carthami teliospores. S. americana, a close relative of yellow starthistle, had not been tested previously. All tests were conducted in a containment greenhouse. Foliage of 19 Cirsium species, 11 safflower cultivars, and S. americana was inoculated with urediniospores and subjected to a 16-h dew period at 18–20 °C. Neither the Cirsium species nor S. americana became infected after foliar inoculations. Compared to foliar infections by P. carthami from safflower in California, only minor infections developed from inoculations with P. jaceae. These were similar to infections observed in earlier studies, and it was not possible to maintain P. jaceae under optimal greenhouse conditions on safflower foliage. Quantitative teliospore inoculations with P. jaceae did not cause infection on safflower hypocotyls, even though large cankers occurred on plants inoculated with P. carthami teliospores. Clear microscopic evidence of infection also was observed in hypocotyls inoculated with P. carthami. These data suggest that native (including rare, threatened, or endangered) Cirsium spp., modern safflower cultivars, and S. americana are not likely to be adversely affected by the use of P. jaceae for biological control of YST. Results from these studies substantiate previous findings and were incorporated in a proposal for permission to use P. jaceae for YST control in California.     

Demographic and environmental influences on life-history traits of isolated populations of the Andean catfish Astroblepus ubidiai

In this study, we examined the influence of demographic and environmental variables on the life histories of six remnant populations of the Andean catfishAstroblepus ubidiai (Actinopterygii; Siluriform) located in isolated refuges, and tested six predictions on the relationships between age at maturity, generation time, population density and juvenile and adult growth and survival rates. The three populations inhabiting the watershed with a direct connection to a major lake (Imbakucha) exhibited later maturity, a longer generation time, higher adult survivorship, a higher adult-to-juvenile survival ratio and lower biomass density than those of the adjacent, non-lacustrine watersheds. Across all six populations, there was a strong correlation between mean age at maturity and both the adult-to-juvenile survival ratio and biomass density. Study populations also showed a pattern of inverse density-dependence associated with Allee effects. We conclude that without limitations in the availability of environmental resources, higher population density can be negatively related to age at maturity. This relationship would be expected to become positive when carrying capacity of the systems is reached. Delayed maturity in the Imbakucha populations may be an adaptive response for increasing juvenile survival under stressful conditions.     

Functional response and progeny production of the Madeira mealybug parasitoid, Anagyrus sp. nov. nr. sinope: The effects of host and parasitoid densities

We tested the hypothesis that control of an herbivorous pest would be improved by providing floral resources for adult natural enemies. The herbivore was euonymus scale, Unaspis euonymi (Comstock) (Homoptera: Diaspididae), a serious pest of woody ornamental plants. The experimental landscape consisted of 3 × 3 m plots, each containing a central bed of Euonymus fortunei (Turcz.) that was infested with the scale. Floral resource plants were cultivars of four species that overlapped in bloom periods to provide a continuous supply of floral resources during summer: Trifolium repens L., Euphorbia epithymoides L., Coreopsis verticillata L. var. ‘Moonbeam,’ and Solidago canadensis L. var. ‘Golden Baby.’ Plots contained either low or high densities of all four species, or no resource plants. Densities of euonymus scale were typically lower in plots containing resource plants than in plots without them. Parasitism by Encarsia citrina (Craw.) (Hymenoptera: Aphelinidae) was rarely influenced by the experimental treatments, flower biomass, whole-plant biomass, or scale density, but in some cases was inversely correlated with density of scales within a generation and in the subsequent generation. Parasitism occasionally reduced densities of scales in plots containing resource plants, but this effect apparently was related to vegetative, not floral qualities of plants. A steady increase in parasitism rate over the three-year course of the experiment across the entire landscape was associated with decreasing density of scales, suggesting a numerical response by the parasitoid population. These findings suggest that the parasitoid is capable of effectively controlling euonymus scale in ornamental landscapes where environmental conditions are favorable.     

Hypocrella panamensis sp. nov. (Clavicipitaceae, Hypocreales): a new species infecting scale insects on Piper carrilloanum in Panama

A new species, Hypocrella panamensis, is described from collections and cultures obtained on Barro Colorado Island, Panama. In order to aid in placement of this fungus, phylogenetic analyses were conducted using LSU (rDNA) sequences. Hypocrella panamensis is characterized by possessing pulvinate stromata with a Lecanicillium-like anamorphic state and superficial perithecia. Hypocrella panamensis consistently grouped in a clade containing Hypocrella nectrioides, H. phyllogena, and H. africana (100 % PP). Most species of Hypocrella possess Aschersonia or Hirsutella anamorphs. Hypocrella panamensis is unique in the genus Hypocrella in possession of a Lecanicillium-like anamorphic state. In its biological habit Hypocrella panamensis is similar to other species in Hypocrella in that it infects and degrades the scale insect, then grows superficially on nutrients that emerge to the plant surface through the stylet wound.     

Chaetomium globosum for reducing primary inoculum of Diaporthe phaseolorum f. sp. meridionalis in soil-surface soybean stubble in field conditions

The potential of an antibiotic-producing isolate of Chaetomium globosum (CgA-1) to suppress Diaporthe phaseolorum f. sp. meridionalis (Dpm) in soybean stubble was studied in field microplots of no-tillage, minimum-tillage, and shallow plowing. Mature soybean stems colonized in vitro with Dpm were spread on the soil surface and C. globosum ascospore suspension, without nutrient supply, was sprayed over the entire plot prior to any tillage operation. Perithecial formation and survival of Dpm in soybean stems, concomitantly with colonization by C. globosum, were monitored for a 180-day period (mid-autumn through winter and mid-spring), which is the normal interval between soybean harvest and sowing. The proportion of soybean stem segments occupied by Dpm and number of perithecia formed decreased linearly with time and showed a strong negative correlation with increase in the occupation by C. globosum. At the end of the study, which coincided with the soybean sowing season, the soybean stubble was free from viable Dpm and was colonized by C. globosum. The effectiveness of C. globosum in eliminating the pathogen from surface-borne residue or harrowed-in residue was similar but much slower than in the shallow-plowed microplots. C. globosum successfully competed with major interfering fungi such as, Trichoderma, Nigrospora, and Fusarium in colonizing the soybean stems above and under the soil surface. The data provide strong evidence for use of the antibiotic-producing isolate of C. globosum to control soybean stem canker disease.     

Biological control of brown rot (Moniliana laxa Ehr.) on apricot (Prunus armeniaca L. cv. Hacıhaliloğlu) by Bacillus, Burkholdria, and Pseudomonas application under in vitro and in vivo conditions

In 2002 and 2003, a study was conducted to determine the effect of bacterial strains, Burkholdria OSU 7, Bacillus OSU 142, and Pseudomonas BA 8, on biological control of brown rot disease (Monilinia laxa Ehr.) on apricot cv. Hacıhaliloğlu in Malatya province of Turkey. Apricot orchard at full blooming stage was inoculated with conidial suspension (1 × 10⁶ spores/ml) of M. laxa Ehr. After inoculation, two apricot trees for each application were treated with each of the three biological control agents (Burkholdria gladii OSU 7, Bacillus subtilis OSU 142, and Pseudomonas putida BA 8) by spraying (1 × 10⁹ cfu/ml) on inoculated branches. Disease incidence was evaluated for untreated (control 1) and four different treatment groups including commercial disease management (control 2, positive control: 3% Bourdox in fall, 50% Cupper at pink flower, 30 g/100 l Corus at first blooming, and 300 g/100 l Captan at last blooming stage) and treatments including each of the three bacterial strains (OSU 7, OSU 142, and BA 8). The results showed that disease incidence for negative control (control 1) was 9.94, which was significantly higher than disease incidence for commercial application (2.57%) or bacterial treatments (2.82–5.00%) in the first year. In 2003, the lowest disease incidence observed in OSU 7 treatment (6.80%), while disease incidence rate for positive control and negative control were 9.45% and 28.46%, respectively. This result may suggest that OSU 7 has potential to be used as biopesticide for effective management of brown rot disease on apricot.     

Evaluation of Beauveria bassiana and B. brongniartii strains and four wild-type fungal species against adults of Bactrocera oleae and Ceratitis capitata

The virulence of two isolates of the hyphomycete fungi, Beauveria bassianaand B. brongniartii, and additional fungal species isolated from diseased Bactrocera oleae pupae and Sesamia nonagrioideslarvae were assessed against adults of the olive fruit fly B. oleae and the Mediterranean fruit fly Ceratitis capitata (Diptera: Tephritidae). Contact and oral bioassays revealed that moderate to high mortality rates for the olive fruit fly occurred when the adults were exposed to conidia of Mucor hiemalis, Penicillium aurantiogriseum, P. chrysogenum and B. bassianaisolates. A strain of M. hiemalis isolated from S. nonagrioides larvae was the most toxic resulting in 85.2% mortality to the olive fruit fly adults. B. brongniartiiand B. bassiana were the most pathogenic to the C. capitataadults causing 97.4 and 85.6% mortality. Metabolites collected from the M. hiemalis and P. chrysogenum isolates were toxic to adults of both species.     

Host preference in Pseudacteon phorid flies: response of P. tricuspis and P. curvatus to black, red and hybrid imported Solenopsis fire ants in multiple choice bioassays

Host preferences in both sexes of Pseudacteon tricuspis Borgmeier (Jaguariuna biotype) and Pseudacteon curvatus Borgmeier (Formosa biotype) and their relative attraction to the imported fire ants (IFA), Solenopsis invicta Buren (red IFA), Solenopsis richteri Forel (black IFA) and S. invicta × S. richteri hybrids (hybrid IFA) were investigated in two separate experiments utilizing multiple choice flight bioassays. The results of both experiments clearly showed that both sexes of the Jaguariuna biotype of P. tricuspis could distinguish among the three IFA species and demonstrated greater preference for hybrid IFA and red IFA. This conclusion is supported by a variety of data collected on the number of fly visits, attack rate, and hovering duration (Experiment 1), and on the number of trapped flies (Experiment 2), which showed that black IFA is the least preferred of the three species. Similar results were recorded for the Formosan biotype of P. curvatus, although the data were not as strongly conclusive. Females of this biotype spent a significantly greater amount of time in hovering mode over red IFA and hybrid IFA compared to black IFA, but the other data were not significant. The red IFA is the natural host of both phorid fly biotypes and our results suggest that both biotypes may have evolved a specialized relationship with red IFA including an ability to discriminate it from related fire ants. These results are discussed in relation to the possible role of fire ant chemicals in mediating host preferences in phorid flies, contributions of male phorid flies to fire ant biocontrol, and the practical implications of the key findings.     

Purification and properties of d-hydantoin hydrolase and N-carbamoyl-d-amino acid amidohydrolase from Flavobacterium sp. AJ11199 and Pasteurella sp. AJ11221

We characterized recombinant d-hydantoin hydrolase (DHHase) and N-carbamoyl-d-amino acid amidohydrolase (DCHase) from Flavobacterium sp. AJ11199 and Pasteurella sp. AJ11221. The DHHases from these two strains showed a wide range of hydrolytic activity for various 5-monosubstituted d-hydantoin compounds, including a very high level activity for d-hydantoin compounds corresponding to d-aromatic amino acids such as d-tryptophan d-phenylalanine and d-tyrosine. The DCHases, in turn, were capable of catalyzing the hydrolysis of various N-carbamoyl-d-amino acids (NCD-A.A.) corresponding to d-aliphatic and d-aromatic amino acids. The combination of these enzymes was found to be applicable for the production of various d-amino acids.     

The phylogenetic placement of the Leucogastrales, including Mycolevis siccigleba (Cribbeaceae), in the Albatrellaceae using morphological and molecular data

The morphology of many hypogeous fungi converges on a homogeneous reduced form, suggesting that disparate lineages are subject to a uniform selection pressure. The primary goal of this study was to evaluate the morphology and infer the phylogeny of the Leucogastrales with Mycolevis siccigleba using a Bayesian methodology. A comprehensive morphological assessment was used for an a priori phylogenetic inference to guide the sequencing effort. All structures except spore ornamentation pointed to the Albatrellaceae as the most likely sister taxon. Polyporoletus sublividus, a close relative of Albatrellus, produces ornamented basidiospores with a similar structure to M. siccigleba basidiospores. The ITS from 30 taxa was used for the molecular phylogenetic analysis. P. sublividus was found sister to Mycolevis. Leucophleps spinispora and L. magnata formed a group sister to the Polyporoletus/Mycolevis group, whereas Leucogaster was polyphyletic with respect to the core of the Leucogastrales and sister to A. caeruleoporus. This relationship was expected as previously undescribed chlamydospores produced by members of Albatrellus had a similar morphology to the basidiospores of L. rubescens.     

The potential of Ulocladium botrytis for biological control of Orobanche spp.

A strain of Ulocladium botrytis isolated from diseased Orobanche crenata shoots caused disease on the parasitic weed in pathogenicity tests. The potential of the fungus to be developed as a mycoherbicide for Orobanche spp. was further investigated. Although the fungus significantly decreased O. crenata germination in vitro by 80%, it did not generally lead to a decreased number of O. crenata shoots or tubercles in inoculated root chambers or pots. However, the number of diseased or dead tubercles and underground shoots was significantly increased compared to the noninoculated treatments. Postemergence inoculation of O. crenata shoots with a conidial suspension resulted in the death of almost all inoculated plants 14 days after application under greenhouse conditions. In preliminary host-range studies, the pathogen caused disease on Orobanche cumana on sunflower whereas on Orobanche aegyptiaca shoots parasitizing tomato only minimal disease symptoms could be detected after postemergence inoculation. Based on the results of our investigations, we conclude that Ulocladium botrytis has only a limited potential to be used as a biocontrol agent against Orobanche spp.     

Biological control of Lygus lineolaris by Peristenus spp. in strawberry

Peristenus digoneutis Loan (Hymenoptera: Braconidae) was introduced to the US for biological control of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), and has since spread through much of the northeast. The purpose of this study was to determine if P. digoneutis and a native congener, Peristenus pallipes (Curtis), parasitize L. lineolaris in strawberry (where it is a key pest), and what factors relate to parasitism levels. During 1997–1999 we monitored parasitism on 17 strawberry farms in 14 counties in eastern and western New York State. We found that in eastern NY (where P. digoneutis has been established since the early 1990s), overall mean parasitism was 19.7% (ranging from 0 to 70%), mostly by P. digoneutis. Mean parasitism was significantly lower (12.3%, ranging from 0 to 58%) in western NY (where P. digoneutis was first recorded in 1999), and was mostly by P. pallipes. P. pallipes parasitism was significantly lower in eastern than western NY, suggesting the potential for competitive interaction with P. digoneutis. The insecticide regime of a farm was an important factor influencing parasitism rate, which was 5- to 6.5-fold higher on organic or casually sprayed farms than on intensely treated farms, though pest density under these three regimes was not significantly different. L. lineolaris density, and parasitism rate in nearby alfalfa and abandoned fields were also significant factors for parasitism in strawberry.     

Morphological and molecular investigations of Paramecium schewiakoffi sp. nov. (Ciliophora, Oligohymenophorea) and current status of distribution and taxonomy of Paramecium spp.

Paramecium schewiakoffi sp. nov. is described from a pond in Shanghai, China. It is a freshwater species belonging to the “aurelia” subgroup of the genus. It is of similar size and shape to P. jenningsi, but has a single large micronucleus of the “chromosomal” morphological type, while P. jenningsi has two smaller micronuclei. The general morphology, morphometric characteristics and nuclear reorganization pattern, a random amplified polymorphic DNA (RAPD) fingerprint pattern, and the small subunit rRNA gene sequence are presented for the species. Comparison of P. schewiakoffi with the other species of Paramecium indicates that it is a valid new species of the genus. Geographical locations reported for many Paramecium species do not support the theory that all ciliates have a cosmopolitan distribution. It is proposed that, in an extension of Jankowski's earlier suggestion, the genus Paramecium should be subdivided into four subgenera: Chloroparamecium, Helianter, Cypriostomum and Paramecium, on the basis of morphometric, biological and molecular differences.     

Development of Encarsia bimaculata (Heraty and Polaszek) (Hymenoptera: Aphelinidae) in Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) nymphs

Encarsia bimaculata was recently described from India as a potentially useful parasitoid of Bemisia tabaci. Its developmental biology was studied in the laboratory at 25–30 °C and 70–75% RH. Results showed that E. bimaculata is a solitary, arrhenotokous, heteronomous, autoparasitoid. Mated females laid eggs internally in B. tabaci nymphs that developed as primary parasitoids. Males developed as hyperparasitoids, either in females of their own species or in other primary aphelinid parasitoids. Superparasitism was common under cage conditions. Both sexes have an egg, three larval instars, prepupal, and pupal stages. Development from egg to adult took 12.70 ± 2.10 days for females and 14.48 ± 2.60 days for males. Individual B. tabaci nymphs were examined for E. bimaculata parasitization using three isozymes: esterase, malate dehydrogenase, and xanthine dehydrogenase. All three isozymes showed differential banding patterns that identified E. bimaculata parasitized or unparasitized B. tabaci nymphs.     

Probing the Drosophila retinal determination gene network in Tribolium (II): The Pax6 genes eyeless and twin of eyeless

The Pax6 genes eyeless (ey) and twin of eyeless (toy) are upstream regulators in the retinal determination gene network (RDGN), which instructs the formation of the adult eye primordium in Drosophila. Most animals possess a singleton Pax6 ortholog, but the dependence of eye development on Pax6 is widely conserved. A rare exception is given by the larval eyes of Drosophila, which develop independently of ey and toy. To obtain insight into the origin of differential larval and adult eye regulation, we studied the function of toy and ey in the red flour beetle Tribolium castaneum. We find that single and combinatorial knockdown of toy and ey affect larval eye development strongly but adult eye development only mildly in this primitive hemimetabolous species. Compound eye-loss, however, was provoked when ey and toy were RNAi-silenced in combination with the early retinal gene dachshund (dac). We propose that these data reflect a role of Pax6 during regional specification in the developing head and that the subsequent maintenance and growth of the adult eye primordium is regulated partly by redundant and partly by specific functions of toy, ey and dac in Tribolium. The results from embryonic knockdown and comparative protein sequence analysis lead us further to conclude that Tribolium represents an ancestral state of redundant control by ey and toy.