Cotesia orobenae (Hymenoptera: Braconidae), a Gregarious Endoparasitoid of Evergestis rimosalis (Lepidoptera: Pyralidae), and Hyperparasitoids in Virginia Brassica Crops

Cotesia orobenae Forbes (Hymenoptera: Braconidae) is a gregarious endoparasitoid of the cross-striped cabbageworm (Evergestis rimosalis Guenée) (Lepidoptera: Pyralidae), a minor pest of Brassica crops in Virginia. Cocoons of C. orobenae were collected, weekly, from field plots of broccoli and cabbage crops in Whitethorne, Montgomery County (VA) during 1989 and 1990. Counts were also made of the eggs and larvae of E. rimosalis to determine the pest′s seasonality. In January 1991, C. orobenae cocoons were collected from the held plots to see if C. orobenae overwinters within cocoons. In May 1993, trap plants bearing E. rimosalis larvae were placed among the field plants weekly to determine if C. orobenae adults are present and ovipositing early in the season. Field counts of E. rimosalis eggs and larvae in 1990 indicated the occurrence of three generations off. rimosalis from June to October, but parasitism of larvae on trap plants by C. orobenae in May 1993 suggested that the parasitoid is present in mid-May. C. orobenae oviposits in first through third instar E. rimosalis and the resulting larvae feed internally on the developing host, exit the fourth (last) instar host, and spin cocoons in a loose mass. Mean brood size of 118 C. orobenae field-collected cocoon masses was 9.2 ± 4.4 cocoons per mass. Out of 1085 cocoons, adult C. orobenae emerged from 77%, hyperparasitoids emerged from 7%, and nothing emerged from 16%. Females accounted for 73% of the emerging C. orobenae adults. Data from field-collected cocoons suggest that C. orobenae enter diapause in late September and overwinter in cocoons. Six pupal hyperparasitoids were recovered: Isdromas lycaenae (Howard) (Ichneumonidae); Conura (=Spilochalcis) torvina (Cresson) and Conura (=Spilochalcis) albifrons (Walsh) (Chalcididae); Hypopteromalus tabacum (Fitch), Catolaccus cyanoideus Burks, and Trichomalopsis viridescens (Walsh) (Pteromalidae).     

Intrinsic competition between the gregarious parasite,Cotesia glomeratus and the solitary parasite.Cotesia rubecula [Hymenoptera: Braconidae] for their host.Artogeia rapae [Lepidoptera: Pieridae]

The solitary endoparasite,Cotesia rubecula, was superior to the gregarious endoparasite,Cotesia glomeratus, in tests of intrinsic competition for their hostArtogeia rapae. When an egg ofC. rubecula was oviposited in the host prior to those ofC. glomeratus, C. rubecula always won the ensuing competition. When an egg ofC. rubecula was laid in the same host within 48 h after oviposition byC. glomeratus, C. rubecula was successful in more than 90% of the interactions. Even when the egg ofC. rubecula was oviposited in the same host 72 to 96 h after oviposition byC. glomeratus, C. rubecula won the competition approximately 33% of the time. The implications of intrinsic competition are discussed in relation to the establishment ofC. rubecula in an ecosystem containingA. rapae andC. glomeratus.      

Differential susceptibility of Cotesia orobenae, a parasitoid of the cross-striped cabbageworm, to commonly used insecticides in Cruciferae

The larval endoparasitoid Cotesia orobenae Forbes (Hymenoptera: Braconidae) is an effective natural enemy of the cross-striped cabbageworm, Evergestis rimosalis (Guenée) (Lepidoptera: Pyralidae), in southwest Virginia. Routine use of insecticides to control Plutella xyostella (L.) and other lepidopterous larvae in commercial plantings of cabbage, cauliflower and broccoli can disrupt populations of C. orobenae, causing localized outbreaks of the cross-striped cabbageworm to occur. Since the use of insecticides is the choice insect pest management tactic of growers in southwest Virginia, we examined the differential susceptibility of this hymenopteran parasitoid to four frequently used pesticides, methomyl (carbamate), permethrin and esfenvalerate (pyrethroids), and Bacillus thuringiensis Berliner subsp. Kurstaki (bacterium). Filter paper dipped into four different concentrations of methomyl, permethrin, and esfenvalerate were left to dry and then exposed to C. orobenae. Two concentrations of B. thuringiensis were also tested by contact bioassay, and an additional test was conducted by mixing B. thuringiensis with honey and water as a food source for the insects. Ten adult parasitoids were exposed to each treatment, replicated 10 times. Significant differences in mortality among insecticides tested (p < 0.05) were found. Most toxic to C. orobenae was permethrin, followed by esfenvalerate and methomyl. The parasitoid was not affected by B. thuringiensis when placed in contact with B. thuringiensis-treated broccoli leaves, or by ingestion of honey mixed with it (p < 0.05).     

Parasites of the spotted stalk borer,Chilo partellus [Lepidoptera: Pyralidae] in South Africa

One egg parasite, 7 larval parasites, 2 pupal parasites and 3 larval hyperparasites were recorded parasitizing the spotted stalk borer,Chilo partellus (Swinhoe) on maize and grain sorghum in South Africa.Trichogrammatoidea lutea Girault [Trichogrammatidae] parasitized eggs ofC. partellus mainly in mid-summer. The larval parasites were active throughout the season with occasional peaks of up to 75% parasitism.Apanteles sesamiae Cameron [Braconidae], proved to be the most abundant larval parasite. It was recorded fromca. 93% of parasitized larvae but its efficiency was reduced by the hyperparasite,Aphanogmus fijiensis (Ferriére) [Ceraphronidae], which reached sometimes up to 100% parasitism on cocoons ofA. sesamiae. The efficiency ofIphiaulax sp. [Braconidae], the 2^nd most abundant larval parasite, was also hindered by the hyperparasite,Eurytoma sp. [Eurytomidae] Pupal parasites were sometimes very abundant reaching up to 100% parasitism without any interference by hyperparasites. The most abundant pupal parasites wereDentichasmias busseolae Heinrich [Ichneumonidae] andPediobius furvus (Gahan) [Eulophidae].      

Effects of nitrogen and foliar biomass on population parameters of cabbage insects

The effects of different nitrogen (N) fertilization rates (0, 45, 90, and 168 kg N/ha), plant nitrogen concentration, and plant biomass on abundance and population growth of diamondback moth, Plutella xylostella (L.), cabbage looper, Trichoplusia ni (Hübner), cabbage budworm, Hellula phidilealis (Walker), imported cabbageworm, Artogeia rapae (L.), and cross-striped cabbageworm, Evergestis rimosalis (Guenée), were investigated in Homestead and Sanford, Florida in 1987. The effects of these factors on the parasitization of P. xylostella were also examined. In Homestead, abundance of most insect pests and parasitized P. xylostella increased with an increase in the level of N applied and with an increase in plant biomass. Similar results were found in Sanford, although results were not consistently significant. Abundance of most insect pests was significantly positively correlated with plant N concentration. Multiple regression analyses indicated that foliar biomass was significantly more important than N fertilization rate and subsequent plant N concentration at predicting abundance of insect pests and parasitized P. xylostella on cabbage.     

Phenology of dormancy in a Virginia population ofMesocyclops edax (Crustacea: Copepoda)

The abundance and age structure of a Virginia, USA population ofMesocyclops edax that undergoes dormancy in an eutrophic lake was investigated from October 1988 through March 1989. During the fall all instars ofM. edax were present in the water column. By early December, only CI-CIV instars were found. M. edax was not found in the water column in January, but sediment cores in February revealed the presence of CIV's and adult females distributed predominantly at 0–3 cm and in lesser abundance at 3–6 cm. NoM. edax were found below 6 cm. By early March animals in the sediments emerged from dormancy and colonized the water column; CIV's and adult females predominated while CV's and adult males were present in small numbers. Dormancy of both juvenile and adult instars within one population is unusual among animal species. Life histories including multiple dormant stages may be more important for the adaptive evolution of cyclopoid copepods than calanoid copepods in freshwater habitats.     

Floral homeotic gene expression defines developmental arrest stages inBrassica oleracea L. vars.botrytis anditalica

Brassica oleracea L. vars.italica (broccoli) andbotrytis (cauliflower) both undergo developmental arrests which result in heading phenotypes. We characterized these arrested tissues at the morphological and molecular levels, and defined the developmental changes that ensue after arrest has been broken. We found that the order of floral organ initiation and the positions of resulting floral organ primordia in this species differed in some respects from that ofArabidopsis, which is a member of the same family, Brassicaceae. We also cloned homologs of theArabidopsis floral homeotic genesAPETALA1 (AP1) andAPETALA3 (AP3) fromB. oleracea and characterized their expression patterns. We found that theAP1 homolog was expressed in some of the meristems of arrest-stage cauliflower, providing evidence that this tissue is florally determined. In broccoli, both the API andAP3 homologs were expressed. However, the spatial pattern of expression of the broccoliAP1 homolog differed from that ofArabidopsis. In addition, we identified a homolog of theCAULIFLOWER (CAL) gene,BoiCAL, from broccoli. The predicted amino acid sequence indicated that theBoiCAL gene product does not contain the mutation thought to be responsible for the early arrest exhibited in cauliflower (Kempin et al. 1995), but contains other changes that might play a role in the broccoli heading phenotype.The GenBank accession numbers for the sequences reported here are: U67451 (Boi2AP1); U67452 (Boi2AP1); U67453 (Boi1AP3); U67455 (Boi2AP3); U67456 (BobAP3); and U67454 (BoiCAL)     

Cover crop mulches influence biological control of the imported cabbageworm (Pieris rapae L., Lepidoptera: Pieridae) in cabbage

Increasing structural complexity within crop fields can provide a way to manipulate pest abundance and biological control in agroecosystems. Here, we examine the effect of cover crop mulches in cabbage on the structure and function of an insect food web, investigating the role of cover crop species, structure, and volatile cues on important interactions. We focused on the imported cabbageworm (Pieris rapae L., Lepidoptera: Pieridae), and three of its natural enemies, the spined soldier bug (Podisus maculiventris (Say), Hemiptera: Pentatomidae), the convergent lady beetle (Hippodamia convergens (Guerin), Coleoptera: Coccinellidae), and the parasitoid, Cotesia rubecula Marshall (Hymenoptera: Braconidae). We measured the abundance of these insects in a field experiment and conducted a natural enemy exclusion cage study to determine the level of biological control of the imported cabbageworm in the field. Our field experiments indicated that cover crop species, but not structure, influenced insect abundance, with significantly more imported cabbageworm and C. rubecula in rye cover crop mulch plots compared to vetch mulch or bare soil plots. In the Y-tube assays we found some evidence that the increased parasitoid abundance did not result in increased parasitism because of interference of the mulch with short-range host finding odor cues. The natural abundance of the two predators was not different among our field plots with different cover crop treatments. Mortality and parasitism of sentinel imported cabbageworm larvae was not different in field cages among the different cover crop mulch treatments, but there was a significant difference among cage types indicating that small natural enemies play an important role in the biological control of this cabbage pest.     

Larval mortality of pine sawflies [Hymenoptera: Diprionidae] in relation to pollution level: A field experiment

Larval mortality ofNeodiprion sertifer (Geoffroy),Diprion pini (L.) andGilpinia pallida (Klug) were studied in field experiments around a factory complex in southwestern Finland. Larval colonies were transferred on the shoots of Scots pines growing at different distances from the emission source. Larval mortality was highest near the factories. InN. sertifer, larval mortality caused by the nuclear polyhedrosis virus was higher and cocoon mortality caused by parasitoids was lower near the pollutant source. The most abundant parasitoid species wereSynomelix scutulata (Hartig) andLamachus eques (Hartig). 16–67% of theN. sertifer, 0–5% of theD. pini and the 73–100% ofG. pallida cocoons contained parasitoids oviposited during the larval period of the sawflies.      

Genetic diversity of dodder (<Emphasis Type="Italic">Cuscuta</Emphasis> spp.) collected from commercial cranberry production as revealed in the<Emphasis Type="Italic">trn</Emphasis>L (UAA) intron

Swamp dodder (Cuscuta gronovii) is a parasitic plant detrimental to cranberries. Observation of emergence of dodder seeds collected from a cultivated cranberry bog in Massachusetts revealed 2 or more peak emergence times during 4 consecutive growing seasons. Molecular methods were used to investigate genetic variation among the emerging dodder seedlings. On emergence, dodder seedlings were collected and analyzed for DNA sequence diversity in thetrnL (UAA) intron, a noncoding region of chloroplast DNA. DNA sequence analysis of 87 seedlings collected during the 1999 and 2000 growing seasons revealed the presence of 2 dodder ecotypes, designated A and B. Comparative DNA sequence analysis indicated that in thetrnL (UAA) intron, the sequence of ecotype A is identical to that ofCuscuta gronovii, whereas the sequence of ecotype B is closest to that ofCuscuta attenuata (99.3% sequence identity; 293 bases considered). ABg/II restriction enzyme cut site was identified that distinguished between thetrnL (UAA) introns of ecotypes A and B. Restriction fragment length polymorphism (RFLP) was used to analyze the sequences of 100 seedlings collected during the growing seasons of 2001 and 2002. Only 10 of the 187 samples were ecotype A, all of which emerged on or before May 7 in the growing seasons. Therefore, the predominant dodder haplotype found in this study may be a close relative ofC. attenuata and notC. gronovii, the common species found in cranberry bogs.     

Impact of flowering buckwheat on Lepidopteran cabbage pests and their parasitoids at two spatial scales

We assessed the potential of annual buckwheat, Fagopyrum esculentum Moench, to lead to improved parasitism of lepidopteran cabbage pests over four years. Pest, parasitism, and hyperparasitism rates were monitored in replicated cabbage plots (12 × 20 m) with or without 3 m wide buckwheat borders from 2000 to 2003. Floral borders did not significantly increase egg, larval, or pupal densities of cabbage looper, Trichoplusia ni (Hübner), imported cabbageworm, Pieris rapae (L.), or diamondback moth, Plutella xylostella (L.). Buckwheat increased parasitism rates by Voria ruralis (Fallen) on T. ni larvae and Cotesia rubecula (Marshall) on P. rapaelarvae over four years. Parasitism by Diadegma insulare (Cresson) on P. xylostella larvae was higher in buckwheat than control plots in the first year, and parasitism by Euplectrus plathypenae (Howard) on T. ni larvae was lower in buckwheat than control plots in the second year. The hyperparasitoid Conura side (Walker) attacked D. insulare all four years, but buckwheat did not affect hyperparasitism rates. The effect of spatial scale on pest densities and parasitism in 2001 was evaluated by comparing plots separated at least 67 m (nearby) versus 800 m apart (isolated). T. ni pupae and P. rapae eggs and pupae were more abundant in plots in closer proximity, whereas P. xylostella densities did not vary by the spatial separation of plots. Tachinids and Pteromalus puparum (L.) attacked more P. rapae in nearby plots. E. plathypenae responded to the treatment × scale interaction, parasitizing more in control than buckwheat when plots were isolated but not when plots were nearby.     

Establishment and Dispersal ofUrophora affinis(Diptera: Tephritidae) andMetzneria paucipunctella(Lepidoptera: Gelechiidae) in Southwestern Virginia

Two European gall-producing insects,Urophora affinisFrfld. (Diptera: Tephritidae) andMetzneria paucipunctella(Zeller) (Lepidoptera: Gelechiidae) were introduced into Virginia in 1986 for biological control of spotted knapweed (Centaurea maculosaLam.). Adults ofU. affinis(n = 2625) andM. paucipunctella(n = 450) were released at two sites in Montgomery County, Virginia, and their populations were monitored yearly by dissecting spotted knapweed flower heads. Beginning in 1992, knapweed samples collected at various distances from the release sites were checked for dispersal.U. affinisis well established and is spreading slowly. The number of larvae per flower head and the seed numbers are inversely related as plants with the greatest number of larvae per spotted knapweed head had the lowest number of seeds. Knapweed density has declined at one of the release sites which had the highest rate of infestation byU. affinis.Establishment of the moth,M. paucipunctella,is less certain as it has been recovered at a very low level from only one site.     

Notes on the biology of the freshwater planarian Dugesia bengalensis (Platyhelminthes Turbellaria Tricladida)

Dugesia bengalensis was described by Kawakatsu (Kawakatsu et al., 1983) from specimens collected in West Bengal. We have been studying populations from many different localities in Santiniketan and adjoining areas of West Bengal and can provide additional biological information.The species is hermaphroditic, and its breeding season was found to occur usually between October and March when the winter temperature falls below 25 °C. Outside of the breeding season, D. bengalensis is capable of asexual reproduction by binary fission (Mahapatra et al., 1987).Development of the reproductive organs appeared to be from neoblasts and other mesenchymal cells and, therefore, to be like that typical of other triclads with the exception that some of the neoblasts used for the reproductive tissue appeared to be derived from the intestinal region (Ghosh, 1988; cf. Teshirogi, 1986).During copulation, the partners were oriented in the same direction and not in a head-to-tail position as has been reported (Hyman, 1945) for some planarians.The oval, stalked cocoons were laid in marshy places, and during the period of summer (usually from April to June) they lay dormant in the sandy soil until the onset of the monsoon rains. Then, typically three or four months after they were laid, the cocoons hatched to yield three or four young, a remarkably low number for freshwater triclads (cf Ball & Reynoldson, 1981).     

Innate responses of the parasitoidsCotesia glomerata andC. rubecula (Hymenoptera: Braconidae) to volatiles from different plant-herbivore complexes

To determine and compare innate preferences of the parasitoid speciesCotesia glomerata andC. rubecula for different plant-herbivore complexes, long-range (1-m) foraging behavior was studied in dual-choice experiments in a wind tunnel. In this study we tested the hypothesis that naive females of the specialistC. rubecula should show more pronounced preferences for different plant-herbivore complexes than females of the generalistC. glomerata. The herbivore species used were the pieridsPieris brassicae, P. rapae, P. Napi, andAporia crataegi and the nonhostsPlutella xylostella andMamestra brassicae. All herbivore species feed mainly on cabbage and wild crucifers, exceptAporia crataegi, which feeds on species of Rosaceae. Both parasitoid species preferred herbivore-damaged plants over nondamaged plants. NeitherC. rubecula norC. glomerata discriminated between plants infested by different caterpillar species, not even between plants infested by host-and nonhost species. Both parasitoid species showed preferences for certain cabbage cultivars and plant species. No differences were found in innate host-searching behavior betweenC. glomerata andC. rubecula. The tritrophic system cabbage-caterpillars-Cotesia sp. seems to lack specificity on the herbivore level, whereas on the plant level differences in attractiveness to parasitoids were found.     

Differences in the envelope proteins ofChlamydia pneumoniae,Chlamydia trachomatis,andChlamydia psittaci shown by two-dimensional gel electrophoresis

Analysis by two-dimensional gel electrophoresis of theN-laurylsarkosinate(Sarkosyl)-insoluble envelope complexes ofl-^[35]S-cysteine-iabeled elementary bodies ofChlamydia pneumoniae strain IOL-207,Chlamydia trachomatis serovar LGV2, D, and F, andChlamydia psittaci strain 6BC showed differences in the molecular charges of chlamydial outer membrane proteins. The apparent isoelectric point (pI) of the major outer membrane protein ofC. pneumoniae strain IOL-207 was 6.4, whereas the pI of the major outer membrane protein of theC. trachomatis andC. psittaci strains differed little from one another, ranging from 5.3 to 5.5. The 60-kDa cysteinerich protein ofC. pneumoniae was the only 60-kDa chlamydial protein with a pI value (5.9) more acidic than that of the corresponding major outer membrane protein. As a general rule, the charges of both the 60-kDa and the lowmolecular-mass (12–15 kDa) cysteine-rich proteins were widely variable, depending on the strain. However, in cach individual strain, the variation of the charge of the 60-kDa protein had a compensatory change in the lowmolecular-mass cysteine-rich protein.     

Stable transformation and regulated expression of an inducible reporter construct inCandida albicans using restriction enzyme-mediated integration

To allow the regulated expression of cloned genes inCandida albicans, a plasmid was constructed using the inducible promoter of theC. albicans MAL2 gene. To demonstrate that theMAL2 promoter could regulate cloned genes placed under its control, a fusion construct was made with the coding sequence of theC. albicans URA3 gene. This plasmid was introduced into a Ura^– strain ofC. albicans using the process of restriction enzyme-mediated integration (REMI). This procedure involves the transformation of theBamHI-linearized plasmid in the presence ofBamHI enzyme. The majority of transformants generated contained insertions of the plasmid at chromosomalBamHI sites. All transformants examined were inducible forURA3 expression, which was determined by growth analysis and by measuring the level ofURA3 gene product activity. The Ura⁺ phenotype of the transformants was stable during growth under nonselective conditions. This system offers the advantages of stable transformation, easy recovery of integrated DNA, and inducible expression of genes inC. albicans.Deceased, December 15, 1995     

Phyletic and evolutionary relationships ofBrachyscome lineariloba (Compositae)

A comparison of karyotypes ofBrachyscome breviscapis (2n = 8),B. lineariloba cytodemes E (2n = 10), B (2n = 12) and C (2n = 16) suggests that these species have a homoelogous basic set of four chromosome pairs, two large pairs and two small, and that theB. lineariloba cytodemes E, B and C are related toB. breviscapis by successive additions of small chromosomes. A pronounced asynchrony of chromosome condensation between these large and small chromosomes has been observed. In the artificial hybrids betweenB. dichromosomatica (2n = 4) ×B. breviscapis, and theB. lineariloba cytodemes, theB. dichromosomatica chromosomes are similar in size and condensation behaviour to the small chromosomes ofB. breviscapis and ofB. lineariloba cytodemes E, B and C. Meiotic pairing in these hybrids also demonstrates the strong affinities between these chromosomes. It is suggested thatB. breviscapis may be of amphidiploid origin between a species with two large early condensing chromosome pairs and another,B. dichromosomatica-like species with two small late condensing pairs. It seems most likely that the additional small and late condensing chromosomes inB. lineariloba cytodemes E, B and C are derived from theB. dichromosomatica-like parent, and that each addition increases vigour, fecundity and drought tolerance, allowing these cytodemes to colonize more open and arid environments. Transmission of the univalents in the quasidiploidB. lineariloba cytodeme E was verified as being via the pollen, and not via the embryo sacs.The cytology ofBrachyscome lineariloba (Compositae, Asteroidae), 10.     

rbcL sequences support exclusion ofRetzia,Desfontainia, andNicodemia from theGentianales

The taxonomic positions ofRetzia, Desfontainia, andNicodemia have been much discussed, and all three genera have been included inLoganiaceae (Gentianales). We have made a cladistic analysis ofrbcL gene sequences to determine the relationships of these taxa toGentianales. Four newrbcL sequences are presented; i.e., ofRetzia, Desfontainia, Diervilla (Caprifoliaceae), andEuthystachys (Stilbaceae). Our results show thatRetzia, Desfontainia, andNicodemia are not closely related toLoganiaceae or theGentianales. Retzia is most closely related toEuthystachys and is better included inStilbaceae. The positions ofDesfontainia andNicodemia are not settled, butDesfontainia shows affinity for theDipsacales s.l. andNicodemia for theLamiales s.l.     

Influences of plant species on life history traits of Cotesia rubecula (Hymenoptera: Braconidae) and its host Pieris rapae (Lepidotera: Pieridae)

The relative suitability of four plants was studied for larvae of Pieris rapae L. and its parasitoid Cotesia rubecula (Marshall). For unparasitized P. rapae, pupal dry weight and egg-pupa growth rate were higher on cabbage, radish and nasturtium than on Indian hedge mustard. Larval developmental rate and size were greatest for C. rubecula when its host was feeding on nasturtium. Wasp survival was not affected by the host insect/plant combination in which the parasitoid developed. These results indicate that the plant on which host larvae feed is an important factor in development of the parasitoid.     

Implications ofrbcL sequence data for higher order relationships of theLoasaceae and the anomalous aquatic plantHydrostachys (Hydrostachyaceae)

Subclass and ordinal relationships ofLoasaceae, a small predominately New World family, are examined usingrbcL sequence data. Sequences were examined for eight of the fifteen genera of theLoasaceae and the morphologically anomalous aquatic genusHydrostachys (Hydrostachyaceae). Parsimony analyses of these sequences, combined with previously publishedrcbL data, indicate thatLoasaceae belong in theCornales, and are the sister group ofHydrangeaceae. This agrees with phylogenies based on chloroplast DNA inverted repeat restriction site, morphological and chemical data. TherbcL trees support the monophyly of theLoasaceae and most generic relationships correspond to current subfamily divisions. TherbcL phylogeny also provides the first suggestion thatHydrostachys is allied with theHydrangeaceae in theCornales.