Pathogenicity of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> and <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> to the tobacco spider mite <Emphasis Type="Italic">Tetranychus evansi</Emphasis>

Seventeen isolates of Metarhizium anisopliae (Metschnikoff) Sorokin and two isolates of Beauveria bassiana (Balsamo) Vuillemin were evaluated for their pathogenicity against the tobacco spider mite, Tetranychus evansi Baker & Pritchard. In the laboratory all the fungal isolates were pathogenic to the adult female mites, causing mortality between 22.1 and 82.6%. Isolates causing more than 70% mortality were subjected to dose–response mortality bioassays. The lethal concentration causing 50% mortality (LC₅₀) values ranged between 0.7×10⁷ and 2.5×10⁷ conidia ml⁻¹. The lethal time to 50% mortality (LT₅₀) values of the most active isolates of B. bassiana and M. anisopliae strains varied between 4.6 and 5.8 days. Potted tomato plants were artificially infested with T. evansi and treated with B. bassiana isolate GPK and M. anisopliae isolate ICIPE78. Both fungal isolates reduced the population density of mites as compared to untreated controls. However, conidia formulated in oil outperformed the ones formulated in water. This study demonstrates the prospects of pathogenic fungi for the management of T. evansi.     

Influence of Temperature on Virulence of Fungal Isolates of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> and <Emphasis Type="Italic">Beauveria bassiana</Emphasis> to the Two-Spotted Spider Mite <Emphasis Type="Italic">Tetranychus urticae</Emphasis>

Twenty-three isolates of Metarhizium anisopliae (Metschnikoff) Sokorin and three isolates of Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales: Clavicipitaceae) were assessed for their virulence against the two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae). Based on the screening results, nine isolates of M. anisopliae and two isolates of B. bassiana were tested for their virulence against young adult (1- to 2-day-old) female T. urticae at constant temperatures of 20, 25, 30 and 35°C. At all temperatures tested, all the fungal isolates were pathogenic to T. urticae but mortality varied with isolates and temperatures. Fungal isolates were more virulent at 25, 30 and 35°C than at 20°C. The lethal time to 50% mortality (LT₅₀) and lethal time to 90% mortality (LT₉₀) values decreased with increased temperature. There were no significant differences in virulence between fungal isolates at 30 and 35°C; however, significant differences were observed at 20 and 25°C.     

Nematicidal activity of culture filtrate of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> against <Emphasis Type="Italic">Meloidogyne hapla</Emphasis>

Culture filtrates of Beauveria bassiana at different concentrations were evaluated for nematicidal activity against the northern root knot nematode (Meloidogyne hapla); bioassays included egg hatching, mortality and infectivity on tomato plants in pots under glasshouse conditions. The percentage mortality and inhibition of hatching of root-knot nematode were directly proportional to the concentration of culture filtrates of B. bassiana. Soil drenching with culture filtrate of B. bassiana significantly reduced nematode population densities in soil and in the roots and subsequent gall formation and egg-mass production by M. hapla under glasshouse conditions.     

Natural Infections Caused by the Fungus <Emphasis Type="Italic">Beauveria bassiana</Emphasis> as a Pathogen of <Emphasis Type="Italic">Musca domestica</Emphasis>in the Neotropic

A survey for entomopathogenic fungi of Musca domestica adults was conducted in poultry houses in La Plata, Buenos Aires province, Argentina, during the years 2002 and 2003. Adult house flies were found infected with the fungus Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes) from field collections, with a natural infected prevalence between 0.4–1.45%. This is the first record of natural infections of house flies caused by B. bassiana for the neotropics. Pathogenicity assays under laboratory conditions showed 94% adult mortality at 14 days post challenge. CIC fellow     

Substrate influence on physiology and virulence of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> acting on larvae and adults of <Emphasis Type="Italic">Tenebrio molitor</Emphasis>

Two isolates of Beauveria bassiana, wild type (wt) and its mutant type (mt) were compared in terms of growth patterns on culture plates containing media based on wheat bran, grasshopper exoskeletons, colloidal chitin or Sabouraud-dextrose agar (SDA). Germination for the mt isolate was up to 33% faster in all media. Influence of media on virulence was determined against larvae and adults of Tenebrio molitor. Mortality higher than 90% was reached for adults after 6 days using conidia from all media. For larvae, a mortality of 80% was reached after 11 days with conidia collected from SDA medium and between 15 and 35% with conidia from other media. In SDA medium, conidial yield was almost ten times higher for the mt isolate compared to the wt isolate; however, virulence traits were similar against either larvae or adults. These results may influence commercial preparations of entomopathogenic fungi based on conidia.     

Isolation of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> (Bals.) Vuill. (Deuteromycotina: Hyphomycetes) from the Chagas disease vector, <Emphasis Type="Italic">Triatoma infestans</Emphasis> (Hemiptera: Reduviidae) in Argentina

A survey for natural entomopathogenic fungi of the Chagas disease vector Triatoma infestans was conducted in five provinces of Argentina since 2001. Nymphs (1.5%) and adults (3.3%) infected with a strain of the fungus Beauveria bassiana were found at Dean Funes, Córdoba province, Argentina. Field collected insects that died in the laboratory were maintained in moist chambers and incubated at 22 °C. Beauveria bassiana from infected insects was cultured on SDAY media. Pathogenicity tests were conducted with a conidial suspension (1 × 10⁷ conidia/ml) of this isolate on T. infestans adults. A mortality rate of 100% was obtained at 15 days post-infection. This is the first record of natural infection of T. infestans by B. bassiana.     

Selection of entomopathogenic fungi for use in combination with sub-lethal doses of imidacloprid: perspectives for the control of the leaf-cutting ant <Emphasis Type="Italic">Atta sexdens</Emphasis><Emphasis Type="Italic">rubropilosa</Emphasis> Forel (Hymenoptera: Formicidae)

In the first part of this study, four isolates of the fungus Beauveria bassiana (Bals.) Vuillemin (LPP1, LPP2, CG05 and CG24) and one isolate of Metarhizium anisopliae (Metsch.) Sorokin (CG46) were tested against adult foragers of Atta sexdens rubropilosa. Ants were allowed to walk on filter paper discs, inside Petri dishes, previously impregnated with 1 ml of a conidia suspension (2 × 10⁷ conidia ml⁻¹), maintained at 80% RH and 26°C for 24 h and subsequently, transferred to sterile Petri dishes, maintained at 23°C, 80% RH, 24 h dark. The mean values of LT₅₀ for LPP2, LPP1, CG46, CG24 and CG05 were 3.5, 3.7, 3.8, 4.2 and 4.4 days, respectively. Control insects for all tests in this study showed less than 10% mortality. Experiments were carried out to test the toxicity of imidacloprid (IMI) to A. sexdens rubropilosa. Mortality was evaluated 10 days following a 24 h exposure to the insecticide. Percent mortality caused by 500, 200, 100 and 10 ppm IMI was 77.8, 56.7, 45.5 and 5.5 respectively. Insects treated with 10 ppm IMI were observed to have reduced locomotor activity 24 h after exposure to the insecticide. The LC₅₀ of IMI was 154.3 ppm. Subsequent tests were carried out to evaluate the combination of a sub-lethal dose of IMI (10 ppm) and infection by CG24 (1 × 10⁷ conidia ml⁻¹). Mortality due to fungal infection alone was 43.3%. Mortality of insects treated with IMI followed by exposure to the fungus was 64.3%. These results indicate that IMI significantly increases the susceptibility of ants to infection by B. bassiana CG24.     

Pathogenicity of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> (Deuteromycetes) and permethrin to <Emphasis Type="Italic">Ixodes scapularis</Emphasis> (Acari: Ixodidae) nymphs

Effectiveness of the entomopathogenic fungus Metarhizium anisopliae, for controlling nymphal Ixodes scapularis, was tested in laboratory and field trials. In the laboratory, M. anisopliae (Metschnikoff) Sorokin strain ESC1 was moderately pathogenic, with an LC₅₀ of 10⁷ spores/ml and induced 70% mortality at 10⁹ spores/ml. In a field study, however, 10⁹ spores/ml M. anisopliae did not effectively control questing I. scapularis nymphs, and significant differences were not detected in pre- and post-treatment densities. For nymphs collected and returned to the laboratory for observation, mortality was low in treatment groups, ranging from 20 to 36%. To assess whether a chemical acaricide would synergistically enhance pathogenicity of the fungus, we challenged unfed nymphal I. scapularis with combinations of M. anisopliae and permethrin, a relatively safe pyrethroid acaricide, in two separate bioassays. Significant interactions between M. anisopliae and permethrin were not observed, supporting neither synergism nor antagonism.     

Insecticidal evaluation of <Emphasis Type="Italic">Beauveria bassiana</Emphasis> engineered to express a scorpion neurotoxin and a cuticle degrading protease

To improve the insecticidal efficacy of the entomopathogen Beauveria bassiana, the fungus was genetically modified with an insect-specific scorpion neurotoxin AAIT and an insect cuticle degrading protease PR1A from another insect pathogen (Metarhizium anisopliae). The wild-type and the transformants were bioassayed against the larvae of Masson’s pine caterpillar Dendrolimus punctatus and the wax moth Galleria mellonella. In comparison to the wild-type strain, engineered isolates took fewer spores to kill 50% of pine caterpillars, 15-fold less for the aaIT single transformant Bb13T and eightfold less for the double transformant Bb13TPR1A, respectively. The median lethal times for Bb13T and Bb13TPR1A were reduced by 40% and 36.7%, respectively against D. punctatus and 24.4% and 20.9%, respectively against G. mellonella. Our data showed that the cotransformation of these two genes produced no synergistic effects on virulence improvement. It is evident from this study that AAIT could be degraded by the protease PR1A when they are expressed together, emphasizing that protein interactions need to be evaluated when working with multiple genes, particularly if they include proteases.     

PCR markers for mutated <Emphasis Type="Italic">S</Emphasis>-haplotypes enable discrimination between self-incompatible and self-compatible sour cherry selections

Tetraploid sour cherry (Prunus cerasus L.) exhibits gametophytic self-incompatibility (GSI) whereby the specificity of self-pollen rejection is controlled by alleles of the stylar and pollen specificity genes, the S-RNase and SFB (S haplotype-specific F-box protein gene), respectively. As sour cherry selections can be either self-compatible (SC) or self-incompatible (SI), polyploidy per se does not result in SC. Instead, the genotype dependent loss of SI in sour cherry is due to the accumulation of non-functional S-haplotypes. The presence of two or more non-functional S-haplotypes within sour cherry 2x pollen renders that pollen SC. We previously determined that sour cherry has non-functional S-haplotypes for the S ₁ -, S ₆ - and S ₁₃ -haplotypes that are also present in diploid sweet cherry (P. avium L.). The mutations underlying these non-functional S-haplotypes have been determined to be structural alterations of either the S-RNase or SFB. Based on these structural alterations we designed derived cleaved amplified polymorphic sequence (dCAPS) markers and S-haplotype specific primer pairs that took advantage of either the length polymorphisms between S-haplotypes, differential S-haplotype sequences, or differential restriction enzyme cut sites. These primer pairs can discriminate among the mutant and wild-type S-haplotypes thereby enabling the identification of the S-haplotypes present in a sour cherry individual. This information can be used to determine whether the individual is either SC or SI. In a sour cherry breeding program, the ability to discriminate between SI and SC individuals at the seedling stage so that SI individuals can be discarded prior to field planting, dramatically increases the program’s efficiency and cost-effectiveness.     

Floral development and systematic position of <Emphasis Type="Italic">Arillastrum</Emphasis>, <Emphasis Type="Italic">Allosyncarpia</Emphasis>, <Emphasis Type="Italic">Stockwellia</Emphasis> and <Emphasis Type="Italic">Eucalyptopsis</Emphasis> (Myrtaceae)

We have investigated the floral ontogeny of Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis (of the eucalypt group, Myrtaceae) using scanning electron microscopy and light microscopy. Several critical characters for establishing relationships between these genera and to the eucalypts have been determined. The absence of compound petaline primordia in Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis excludes these taxa from the eucalypt clade. Post-anthesis circumscissile abscission of the hypanthium above the ovary in Stockwellia, Eucalyptopsis and Allosyncarpia is evidence that these three taxa form a monophyletic group; undifferentiated perianth parts and elongated fusiform buds are characters that unite Stockwellia and Eucalyptopsis as sister taxa. No floral characters clearly associate Arillastrum with either the eucalypt clade or the clade of Stockwellia, Eucalyptopsis and Allosyncarpia.We gratefully acknowledge Clyde Dunlop and Bob Harwood (Northern Territory Herbarium) for collecting specimens of Allosyncarpia, and Bruce Gray (Atherton) for collecting specimens of Stockwellia. The Australian National Herbarium (CANB) kindly lent herbarium specimens of Eucalyptopsis for examination. This research was supported by a University of Melbourne Research Development Grant to Andrew Drinnan.     

<Emphasis Type="Italic">Phuphanochloa,</Emphasis> a new bamboo genus (<Emphasis Type="Italic">Poaceae</Emphasis>: <Emphasis Type="Italic">Bambusoideae</Emphasis>) from Thailand

Summary  A new monotypic bamboo genus Phuphanochloa (Poaceae: Bambusoideae) from north-eastern Thailand is described, together with a new species, P. speciosa.     

The <Emphasis Type="Italic">Caenorhabditis</Emphasis><Emphasis Type="Italic">briggsae</Emphasis> genome contains active <Emphasis Type="Italic">CbmaT1</Emphasis> and <Emphasis Type="Italic">Tcb1</Emphasis> transposons

The maT clade of transposons is a group of transposable elements intermediate in sequence and predicted protein structure to mariner and Tc transposons, with a distribution thus far limited to a few invertebrate species. We present evidence, based on searches of publicly available databases, that the nematode Caenorhabditis briggsae has several maT-like transposons, which we have designated as CbmaT elements, dispersed throughout its genome. We also describe two additional transposon sequences that probably share their evolutionary history with the CbmaT transposons. One resembles a fold back variant of a CbmaT element, with long (380-bp) inverted terminal repeats (ITRs) that show a high degree (71%) of identity to CbmaT1. The other, which shares only the 26-bp ITR sequences with one of the CbmaT variants, is present in eight nearly identical copies, but does not have a transposase gene and may therefore be cross mobilised by a CbmaT transposase. Using PCR-based mobility assays, we show that CbmaT1 transposons are capable of excising from the C. briggsae genome. CbmaT1 excised approximately 500 times less frequently than Tcb1 in the reference strain AF16, but both CbmaT1 and Tcb1 excised at extremely high frequencies in the HK105 strain. The HK105 strain also exhibited a high frequency of spontaneous induction of unc-22 mutants, suggesting that it may be a mutator strain of C. briggsae.     

Sociality in <Emphasis Type="Italic">Euglossa</Emphasis> (<Emphasis Type="Italic">Euglossa</Emphasis>) <Emphasis Type="Italic">viridissima</Emphasis> Friese (Hymenoptera,Apidae, Euglossini)

Euglossa viridissima is an orchid bee that forms both solitary and multiple female nests, making it a suitable species for the study of factors leading to diverse degrees of sociality in Euglossines. We conducted observations in eight reused nests (where a first generation of bees had been produced) kept in artificial boxes from the Yucatan Peninsula, Mexico. Five nests were reused (reactivated) by a single female (SFN), two nests reused by a mother and one daughter (MFN1) and one nest reused by the mother and two daughters (MFN2). No single nest was reactivated by unrelated females. The number of foraging trips, their duration and the duration of cell provisioning was not different between SFN and MFN. The overall production of cells per female was not different either between both types of nest. However, in MFN although all females did lay eggs, there was a reproductive skew in favor of the mother (95 and 45% of the brood produced in MFN1 and MFN2 respectively). She showed reproductive control of her daughters through oophagy and displaying threatening behavior when the daughters tried to open a cell where she had laid an egg. Brood losses to parasites (Anthrax sp. (Bom-byliidae) and Hoplostelis bivittata (Megachilidae)) were only found in SFN which possibly reflects and advantage of MFN in this respect. Our results coupled with other studies in Euglossa, reveal that a wide range of social behaviors occur in this genus, from solitary and communal to primitive reproductive division of labor. Multiple factors involving different levels of pressure imposed by food availability and parasites may favor such a diverse range of nesting behaviors. Interestingly, female associations in E. viridissima seem a result of kin selection that is enforced by coercion from mother females on their daughters. More studies are needed to shed light upon the social organization of Euglossa and other Euglossines and on their phylogenetic relationships in order to trace the origins of eusociality in Apidae. Received 12 February 2008; revised 25 June 2008; accepted 17 July 2008.     

The Madagascan genus <Emphasis Type="Italic">Vaughania</Emphasis> is reduced to synonymy under <Emphasis Type="Italic">Indigofera</Emphasis> (<Emphasis Type="Italic">Leguminosae-Papilionoideae-Indigofereae</Emphasis>)

Summary  Eleven species comprising the Madagascan genus Vaughania are subsumed within the large pantropical genus Indigofera. Six new combinations are made; the remaining species were originally described in Indigofera.     

Conidia production by <Emphasis Type="Italic">Beauveria bassiana</Emphasis> (for the biocontrol of a diamondback moth) during solid-state fermentation in a packed-bed bioreactor

Conidia of Beauveria bassiana CS-1, which have the potential for the control of the diamondback moth (Plutella xylostella), were produced by solid-state fermentation (SSF) using a packed-bed bioreactor with rice straw and wheat bran. As the packing density and the bed height were increased, the production of conidia decreased. In a packed-bed bioreactor under no aeration and no addition of polypropylene (PP) foam (control), the total average of conidia was 4.9 × 10⁸ g^-1. The production of conidia was affected more by the addition of PP foam as an inert support than forced aeration and was approx. 23 times higher than that of the control. The total average of conidia produced by B. bassiana was 1.1–1.2 × 10¹⁰ g^-1 . Revisions requested 6 September 2004/2 November 2004; Revisions received 1 November 2004/8 December 2004     

Life history of the predatory mites <Emphasis Type="Italic">Neoseiulus paspalivorus</Emphasis> and <Emphasis Type="Italic">Proctolaelaps bickleyi</Emphasis>, candidates for biological control of <Emphasis Type="Italic">Aceria guerreronis</Emphasis>

The eriophyoid mite Aceria guerreronis Keifer (Eriophyidae), commonly called the coconut mite, is a key pest of coconut fruits. Surveys conducted on coconut palms in Brazil revealed the predatory mites Neoseiulus paspalivorus DeLeon (Phytoseiidae) and Proctolaelaps bickleyi Bram (Ascidae) as the most commonly associated natural enemies of A. guerreronis on coconut fruits. However, virtually nothing is known about the life history of these two predators. We conducted laboratory experiments at 25 ± 0.1°C, 70–90% RH and 12:12 h L:D photoperiod to determine the life history characteristics of the two predatory mites when feeding on A. guerreronis and other potential food sources present on coconut fruits such as Steneotarsonemus furcatus DeLeon (Tarsonemidae), coconut pollen and the fungus Rhizopus cf. stolonifer Lind (Mucoraceae). In addition, the two-spotted spider mite Tetranychus urticae Koch (Tetranychidae) was tested for its suitability as prey. Both predators, N. paspalivorus and P. bickleyi, thrived on A. guerreronis as primary food source resulting in shorter developmental time (5.6 and 4.4 days, respectively), higher oviposition rate (1.7 and 7.0 eggs/female/day, respectively) and higher intrinsic rate of increase (0.232 and 0.489 per female/day, respectively) than on any other diet but were unable to develop or lay eggs when fed T. urticae. Coconut pollen and S. furcatus were adequate alternative food sources for N. paspalivorus and Rhizopus for P. bickleyi. We discuss the relevance of our findings for natural and biological control of the coconut mite A. guerreronis.     

Toxicity and kinetics of spinosad in different developmental stages of the endoparasitoid <Emphasis Type="Italic">Hyposoter didymator</Emphasis> (Hymenoptera: Ichneumonidae) and its host <Emphasis Type="Italic">Spodoptera littoralis</Emphasis> larvae (Lepidoptera: Noctuidae)

The toxicity of spinosad was evaluated using the RaPID Assay^® Spinosad immunosorbent assay in different developmental stages of the parasitoid, Hyposoter didymator, and in its host, fourth-instar larvae of the cotton leafworm Spodoptera littoralis. Spinosad was applied directly to pupae and adults of H. didymator (ingestion or topical application) or to the immature stages of the parasitoid via the host larvae. Low amounts of spinosad were recovered from S. littoralis host larvae after topical treatment, and the compound was mainly retained in the hemolymph. Amounts of spinosad detected in third-instar larvae of H. didymator, pulled out from the hemolymph of parasitized S. littoralis larvae, were 85 pg (3.57 ng a.i./g body weight) in dead larvae, and 82 pg (3.42 ng a.i./g body weight) in alive individuals. After topical treatment of H. didymator cocoons, most of the compound was retained in the silken cocoon, preventing contamination of the pupa. Also in the parasitoid adults, relatively low amounts of spinosad were accumulated in the body overall, but half of all the insecticide recovered was found in the ovaries. The kinetic results obtained help to better understand the toxicity of spinosad in the complex S. littoralis–H. didymator, and to ascertain the compatibility between spinosad and the parasitoid for optimizing the control of lepidopteran pests.     

<Emphasis Type="Italic">Funastrum rupicola</Emphasis> (<Emphasis Type="Italic">Apocynaceae:</Emphasis><Emphasis Type="Italic">Asclepiadoideae</Emphasis>), a new species from Bolivia

Summary   Funastrum rupicola Goyder, a new species of Apocynaceae: Asclepiadoideae from Bolivia, is described and illustrated. The conservation status of this species is assessed.