Host specificity of Asian parasitoids for potential classical biological control of <Emphasis Type="Italic">Drosophila suzukii</Emphasis>

The Asian spotted wing drosophila, Drosophila suzukii, has recently become a serious pest of soft fruits in Europe. Classical biological control through the introduction of larval parasitoids from its native range in Asia is presently being considered. However, host specificity of potential biological control agents has to be determined to avoid releasing species that may have unintended non-target impacts. Larvae of six different European non-target fly species and the target D. suzukii were exposed either on diet or blueberries to three Asian larval parasitoids, Asobara japonica, Leptopilina japonica, and Ganaspis cf. brasiliensis, and one European species, Leptopilina heterotoma. Asobara japonica showed the lowest specificity, attacking and developing in all Drosophilidae. Leptopilina japonica successfully parasitized two non-target Drosophilidae, D. melanogaster and D. subobscura, with one singly progeny emerging from D. immigrans. Ganaspis cf. brasiliensis had the highest level of specificity but variations occurred between two geographical populations tested. A Japanese population was strictly specific to D. suzukii, whereas another population from China parasitized D. suzukii, D. melanogaster and sporadically D. subobscura. The European L. heterotoma successfully developed in D. melanogaster, D. subobscura and occasionally in D. immigrans, but nearly all eggs and larvae in D. suzukii were encapsulated. These results show that Ganaspis cf. brasiliensis is the species with the highest potential for biological control, but more studies are needed on its taxonomic status and the existence of biotypes or cryptic species varying in their specificity before field releases can be conducted in Europe.     

Dietary yeast affects preference and performance in <Emphasis Type="Italic">Drosophila suzukii</Emphasis>

Yeasts play an important role in nutrition physiology and host attraction of many Drosophila species, and associations with various yeast species are documented for several drosophilid flies. The pest Drosophila suzukii (Matsumura) has a predominant association with the yeast Hanseniaspora uvarum. However, research has not been conducted on the nutritional physiology of the yeasts associated with D. suzukii (spotted wing drosophila). Therefore, in this study, we determined whether dietary yeast was nutritionally relevant and whether yeast species closely associated with D. suzukii positively affected life-history traits. Our results confirm a crucial role of dietary yeast in the larval development and survival of D. suzukii. Furthermore, we found specific effects of the closely associated yeast species H. uvarum and Candida sp. on larval survival. Observations of the egg-laying behaviour of D. suzukii on cherry fruits artificially colonised with different yeast species revealed that the number of eggs laid increased on fruits colonised with Candida sp. and Saccharomyces cerevisiae.     

First record of the invasive pest <Emphasis Type="Italic">Drosophila suzukii</Emphasis> in Ukraine indicates multiple sources of invasion

Drosophila suzukii, commonly known as the spotted-wing Drosophila, is an invasive polyphagous fruit pest, which has emerged as a global threat to agriculture in the Americas and in Europe. Due to the rapid spread, great economic losses and its pest behavior, D. suzukii represents a powerful model for invasion biology and pest management studies. However, its current European distribution, invasion routes and levels of genetic diversity in populations of D. suzukii are poorly understood. We present the first report of D. suzukii from Ukraine, with the invasion likely occurring close to 2014. The pattern of genetic variation at cytochrome oxidase I among D. suzukii populations from Europe, USA and Asia reveals comparatively high genetic diversity in the Ukrainian population of this pest species, suggesting a complex invasion scenario from multiple sources. Further monitoring patterns of genetic variation across space and time, to better understand the invasion routes of this invasive insect pest, will be an essential part for developing successful pest management strategies.     

Large-scale spatial dynamics of <Emphasis Type="Italic">Drosophila suzukii</Emphasis> in Trentino,Italy

Drosophila suzukii (Diptera: Drosophilidae) is an invasive alien species devastating soft fruit crops in newly invaded territories. Little is known about the importance and potential of long-distance dispersal at a regional scale. The goal of this work is to investigate D. suzukii dispersal ability during different times of the season, and along an elevational gradient in a mountain valley in Trentino Province, Italy. We employed a mark–release–recapture strategy using protein markers. Flies were recaptured using fruit-baited traps. The protein-marked flies were positively identified using ELISA procedure. Additional microsatellite analyses were performed on D. suzukii collected during autumn at different elevations to characterize the population structure. Results suggest that a portion of the local D. suzukii population moves from low to high elevations during spring and summer and travels back to low elevations in autumn. Genetic analysis further revealed that samples collected during autumn at different elevations belong to the same population. These results show that D. suzukii are able to fly up to about 9000 m away from the marking point and that seasonal breezes likely facilitate long-distance movement. We suggest that these migrations have multiple functions for D. suzukii, including conferring the ability to exploit gradual changes of temperature, food, and ovipositional resources in spring and autumn, as well as to assist in the search for suitable overwintering sites in late autumn. Our findings help to unveil the complex ecology of D. suzukii in Italian mountainous regions and provide important clues for improving the efficacy of integrated pest management control techniques to combat this pest.     

<Emphasis Type="Italic">Drosophila suzukii</Emphasis> flight performance reduced by starvation but not affected by humidity

Drosophila suzukii is widely studied because of its status as a global pest of berries and soft fruits. Environmental conditions and access to food resources impact the physiology and fitness of D. suzukii; these factors could also affect dispersal. Flight mills are a convenient tool for measuring and comparing the flight performance of insects. In this study, two experiments examined the effects of diet and humidity on D. suzukii flight performance using custom-built flight mills, and a third experiment compared the energy reserves of D. suzukii flown or not flown on flight mills. Over all flight assays, the median flight distance and duration were 27.16 m and 2.37 min, respectively, and the mean flight velocity was 0.18 m/s. The maximum flight distance and duration by an individual were 1.75 km and 2.35 h, respectively. Drosophila suzukii provisioned with blossoms, fruits, or standard laboratory diets flew farther distances and longer durations than starved flies. While starvation was associated with reduced flight performance, there were no observed differences between diet types. It remains unclear whether D. suzukii consistently use lipids, glycogen, sugar, or another energy source for flight because tethered individuals may not have flown enough to deplete energy reserves. Humidity did not affect flight performance of D. suzukii within a?~?2 h test period. These data indicate that most D. suzukii are likely to remain within limited area (e.g., within a field) but that some individuals can disperse long distances (field to field spread).     

Successful <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of <Emphasis Type="Italic">Populus tomentosa</Emphasis> with apple <Emphasis Type="Italic">SPDS</Emphasis> gene

The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase （SPDS） genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu＇med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR.     

Effects of <Emphasis Type="Italic">Verticillium nonalfalfae</Emphasis> on <Emphasis Type="Italic">Ailanthus altissima</Emphasis> and associated indigenous and invasive tree species in eastern Austria

The naturally occurring Verticillium nonalfalfae shows promise for biocontrol of the highly invasive Tree of Heaven (Ailanthus altissima), but might also bear a risk for non-target tree species. In this study, we conducted inoculations on potted seedlings of A. altissima as well as on eight indigenous and two invasive tree species associated with Tree of Heaven in Austria. Although vascular discolourations developed in all inoculated tree species, V. nonalfalfae was reisolated from Ailanthus and eight of the ten non-target-species, whereas typical disease symptoms and mortality only occurred on A. altissima. Results confirmed high susceptibility (S) of A. altissima to V. nonalfalfae but indicated tolerance (T) of Acer campestre, Acer pseudoplatanus and Quercus robur, possible resistance (PR) of Fraxinus excelsior, Populus nigra, Tilia cordata, Ulmus laevis and Ulmus minor and resistance (R) of Fraxinus pennsylvanica and Robinia pseudoacacia to this potential biocontrol agent. Results from seedling inoculations were confirmed by cursory field observations in Ailanthus-inoculated forest stands, where admixed A. campestre, A. pseudoplatanus, F. excelsior, Populus alba, R. pseudoacacia and U. laevis canopy trees remained asymptomatic, while mortality was induced in Ailanthus.     

Insecticidal properties of <Emphasis Type="Italic">Thymus persicus</Emphasis> essential oil against <Emphasis Type="Italic">Tribolium castaneum</Emphasis> and <Emphasis Type="Italic">Sitophilus oryzae</Emphasis>

Red flour beetle Tribolium castaneum (Herbst) and rice weevil Sitophilus oryzae (L.) are considered to be the major insect pests in storage. Essential oils from aromatic plants are recognized as proper alternatives to fumigants. Thymus persicus (Ronniger ex Rech. f.) is one of these plants that have medicinal properties and is indigenous to Iran. The essential oil was obtained from aerial parts of the plant and analyzed by GC and GC–MS. Carvacrol (44.69%) and thymol (11.05%) were the major constituents of the oil extracted. In this experiment, fumigant toxicity of the essential oil was studied against T. castaneum, S. oryzae at 27 ± 1°C and 60 ± 5% RH in dark condition. The adult insects were exposed to the concentrations of 51.9, 111.1, 207.4 and 370.4 μl/l air to estimate median lethal time (LT₅₀) values. The fumigant toxicity was increased in response to increased essential oil concentrations. The LT₅₀ values at the lowest and the highest concentrations tested were ranged from 28.09 to 13.47 h for T. castaneum, and 3.86 to 2.30 h for S. oryzae. It was found that S. oryzae adults were much more susceptible to the oil than T. castaneum. After 24 h of exposure, the LC₅₀ values (95% fiducial limit) for T. castaneum and S. oryzae were estimated to be 236.9 (186.27–292.81) and 3.34 (2.62–4.28) μl/l air, respectively. These results suggest that T. persicus essential oil merits further study as potential fumigant for the management of these stored-product insects.     

<Emphasis Type="Italic">Ophiostoma</Emphasis> species associated with bark beetles infesting three <Emphasis Type="Italic">Abies</Emphasis> species in Nikko,Japan

Ophiostoma species were isolated from bark beetles and Abies mariesii, A. veitchii and A. homolepis attacked by the beetles in Nikko, Tochigi, central Honshu, Japan. One to two Ophiostoma species were frequently isolated from each species of bark beetle. Ophiostoma subalpinum was the most common associate of Cryphalus montanus. Ophiostoma sp. B as well as O. subalpinum was a common fungus associated with Polygraphus proximus. Ophiostoma europhioides was isolated from Dryocoetes hectographus and D. autographus as one of the common associates. Ophiostoma sp. J and Ophiostoma sp. S were frequently isolated from D. autographus and D. striatus, respectively. These fungi seem to have specific relationships with particular bark beetles. Ophiostoma sp. B, Ophiostoma sp. J and Ophiostoma sp. S have unique morphological characteristics and appear to be new species. Five trees of A. veitchii, approximately 43 years old, were inoculated with five Ophiostoma species to assess the relative virulence of the fungi. Ophiostoma subalpinum, Ophiostoma sp. B, and O. europhioides had relatively higher virulence than the other species studied.     

Identification of attractive blend for spotted wing drosophila, <Emphasis Type="Italic">Drosophila suzukii</Emphasis>, from apple juice

Drosophila suzukii, commonly known as the spotted wing drosophila (SWD), is an exotic fruit fly from Southeast Asia that was introduced to the temperate regions of North America and Europe in 2008. It attacks a wide variety of fruits and has become a devastating pest of soft-skinned fruit crops. Due to the rapid spread of SWD across the newly invaded continents, fresh fruit markets have a zero-tolerance policy regarding D. suzukii infestation. Specific and efficient D. suzukii detection tools are urgently needed so that farmers can deliver timely management interventions to meet market demands. Since SWD is known to be attracted to damaged and rotting fruits, headspace volatiles from fresh and fermented apple juices were collected and analyzed by gas chromatography–mass spectrometry. Special attention was given to the compounds produced and/or enriched during the fermentation process. After performing a series of laboataory and field tests, we identified a quinary blend, which is more efficient and selective for D. suzukii than the currently standard apple cider vinegar and commercially available SWD lure under field conditions. Identification of SWD attractant will help growers accurately detect D. suzukii adult infestations in orchards, thereby allowing for timely pest management interventions while reducing conventional insecticidal usage to protect our crops, environment, and ecosystem.     

Aboveground biomass equations for individual trees of <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>, <Emphasis Type="Italic">Chamaecyparis obtusa</Emphasis> and <Emphasis Type="Italic">Larix kaempferi</Emphasis> in Japan

Generic equations are proposed for stem, branch and foliage biomass of individual trees in even-aged pure stands of Cryptomeria japonica, Chamaecyparis obtusa and Larix kaempferi. Biomass data was collected from a total of 1,016 individual trees from 247 stands throughout Japan, and five regression models were assessed by root mean square error, mean bias, fit index (FI), and AIC. The results show that a power equation using diameter at breast height (dbh) and height is the most suitable for all species and components. This equation is more accurate than the familiar power equation that uses ‘dbh² height’, and it expresses the greater volume of branch and foliage mass of trees with a lower height/diameter ratio. A power equation using dbh is more reasonable for models with dbh as the only independent variable and more accurate than a power equation using ‘dbh² height’ for estimating branch and foliage mass. Estimating error for branch and foliage mass is larger than that for stem mass, but the entire aboveground biomass can be estimated with an error of less than 19%, except in the case of small trees with dbh less than 10 cm.     

Bioremediation of CCA-treated wood by brown-rot fungi <Emphasis Type="Italic">Fomitopsis palustris</Emphasis>, <Emphasis Type="Italic">Coniophora puteana</Emphasis>, and <Emphasis Type="Italic">Laetiporus sulphureus</Emphasis>

This study evaluated oxalic acid accumulation and bioremediation of chromated copper arsenate (CCA)-treated wood by three brown-rot fungi Fomitopsis palustris, Coniophora puteana, and Laetiporus sulphureus. The fungi were first cultivated in a fermentation broth to accumulate oxalic acid. Bioremediation of CCA-treated wood was then carried out by leaching of heavy metals with oxalic acid over a 10-day fermentation period. Higher amounts of oxalic acid were produced by F. palustris and L. sulphureus compared with C. puteana. After 10-day fermentation, oxalic acid accumulation reached 4.2 g/l and 3.2 g/l for these fungi, respectively. Fomitopsis palustris and L. sulphureus exposed to CCA-treated sawdust for 10 days showed a decrease in arsenic of 100% and 85%, respectively; however, C. puteana remediation removed only 18% arsenic from CCA-treated sawdust. Likewise, chromium removal in F. palustris and L. sulphureus remediation processes was higher than those for C. puteana. This was attributed to low oxalic acid accumulation. These results suggest that F. palustris and L. sulphureus remediation processes can remove inorganic metal compounds via oxalic acid production by increasing the acidity of the substrate and increasing the solubility of the metals.An erratum to this article can be found at     

Chemical alterations induced by <Emphasis Type="Italic">Pycnoporus sanguineus</Emphasis>/<Emphasis Type="Italic">Aspergillus flavipes</Emphasis> co-cultures in wood from different tree species

Chemical alterations following inoculation of Acacia mearnsii, Eucalyptus dunnii, E. grandis, and E. macarthurii with a Pycnoporus sanguineus/Aspergillus flavipes co-culture were investigated. Several wood chemical parameters were measured using standard methods from the pulp and paper industry. The data were described and analyzed using univariate as well as multivariate statistical techniques. Boxplots and in particular biplots show clearly how the chemical composition of each tree species was differently affected by the co-culture. Lignin content was significantly decreased in A. mearnsii, while E. dunnii showed a decrease in cellulose content. The results, therefore, indicate that the manner in which wood is degraded by a specific fungal co-culture depends on the tree species involved. This phenomenon should be considered when selecting fungi for bio-pulping.     

Enzymatic saccharification and ethanol production of <Emphasis Type="Italic">Acacia mangium</Emphasis> and <Emphasis Type="Italic">Paraserianthes falcataria</Emphasis> wood,and <Emphasis Type="Italic">Elaeis guineensis</Emphasis> trunk

We examined the saccharification and fermentation of meals from Acacia mangium wood, Paraserianthes falcataria wood, and Elaeis guineensis trunk. The levels of enzymatic hydrolysis of cellulose and ethanol production were highest for P. falcataria wood and lowest for A. mangium wood. Ultrasonication pretreatment of meal further increased the rates of hydrolysis and ethanol production in meal from P. falcataria wood. Through this pretreatment, hemicelluloses (xylan and xyloglucan) and cellulose were released in the meal from P. falcataria wood. Loosening of hemicellulose associations can be expected to make P. falcataria wood more useful for bioethanol production.     

The behaviour of the species mingling index<Emphasis Type="Italic"> M</Emphasis><Subscript><Emphasis Type="Italic">sp</Emphasis></Subscript> in relation to species dominance and dispersion

Ecosystem conservation plays an increasingly important role in forest management. The value for such conservation has been directly linked to high diversity of species and structure within systems. In the past, forestry inventory practices have concentrated on aspects that were primarily required for timber management. Now, however, much interest has arisen in measures that will indicate higher or lower diversity. The interspersion of trees of different species reflects diversity at a local level and may serve as an important indicator for the simplification of a forest or woodland system. This article shows that the mingling index M_sp is robust in describing the spatial dispersion of plants; some guidelines for interpreting M_sp are provided.     

Effectiveness of Neem (<Emphasis Type="Italic">Azadirachta indica</Emphasis>) insecticides against Brassica pod midge (<Emphasis Type="Italic">Dasineura brassicae</Emphasis> Winn<Emphasis Type="Italic">.</Emphasis>)

The efficiency of the botanical insecticide (BI) NeemAzal T/S (containing 1% Azadirachtin A), on the basis of azadirachtin applied in a dose of 20 g a.i. ha⁻¹ against Brassica pod midge (Dasineura brassicae), has been monitored for 4 years. The biological efficiency of BI was compared with the efficiency of some synthetic insecticides. It was ascertained that BI was very efficient in decreasing the number of damaged oilseed rape pods (ranging from 56.5 to 85.9% compared to untreated plants) and its efficiency was comparable with synthetic insecticides based on Chloronicotinyl (Thiacloprid) and Neonicotinoid (Acetamiprid). BI’s efficiency was, in some years, even significantly higher compared to pyrethroid (λ-cyhalothrin). The high biological efficiency of azadirachtin that we ascertained was significantly reflected in increasing potential crop yields. The yield increase of azadirachtin ranged between 9.3 and 19.4% compared to the control sample. Azadirachtin showed the highest yield for the whole time of experimentation, and in some years the yield increase was significantly higher compared to some synthetic agents.     

<Emphasis Type="Italic">Vicia faba</Emphasis> plants respond to oviposition by invasive <Emphasis Type="Italic">Halyomorpha halys</Emphasis> activating direct defences against offspring

The invasive stink bug Halyomorpha halys is established in many European and American agro-ecosystems, where it causes severe crop losses. Potential control measures might include enhancement of plant defences. When attacked by herbivorous insects that oviposit on it, the plant may respond by priming direct defences, which might affect the development of future brood. Halyomorpha halys attacks numerous plant species in the invaded areas. Here, we investigated whether Vicia faba plants challenged by H. halys females can impair the development of its offspring through the activation of induced direct defences. We measured the weight and dimension of nymphs that developed on oviposition-experienced plants after 7 and 17 days from hatching. Nymphs that developed on oviposition-experienced plants weighed less compared to those that developed either on control plants or on plants solely subjected to H. halys feeding, and third instars showed shorter dimensions (tibia length). In addition, when oviposition-experienced plants were attacked by nymphs, higher and more rapid expression of two jasmonic acid-dependent genes (cysteine proteinase inhibitor gene and NAI1) was detected, possibly due to a mechanism of priming. Increased expression of the salicylic acid-responsive PR1 gene was also detected in egg-experienced plants, although the response was delayed compared to JA-dependent genes. Our results suggest that V. faba plants recognize H. halys oviposition as a warning signal and pre-activate defences against future nymphal herbivory.     

Efficacy of low temperatures for the control of all life stages of <Emphasis Type="Italic">Plodia interpunctella</Emphasis> and <Emphasis Type="Italic">Liposcelis bostrychophila</Emphasis>

The insecticidal effect of low temperatures for the control of all life stages of two stored-product insects, Plodia interpunctella and Liposcelis bostrychophila, was evaluated under laboratory conditions. The temperatures tested were 0, ??5, ??10 and ??15 °C, and, at these temperatures, the insects were exposed for 2, 4 and 8 h and also for 1, 2, 3 and 7 days. Regarding P. interpunctella, the most cold-tolerant life stage was larvae, given that 2 days of exposure were needed for complete (100%) mortality at ??10 °C. Moreover, all larvae were dead at ??15 °C even after 2 h of exposure. Eggs of P. interpunctella were susceptible to cold, as mortality was complete after 7, 1 day, 2 and 2 h at 0, ??5, ??10 and ??15 °C, respectively. L. bostrychophila was by far less susceptible than P. interpunctella. For this species, adults were still alive even after 7 days at ??10 °C, while complete mortality was achieved only after 1 day at ??15 °C. Eggs of L. bostrychophila were the most cold-tolerant life stage, as survival was observed even after 3 days at ??15 °C. The results of the present study provide the first data set on which all life stages of P. interpunctella and L. bostrychophila are evaluated for their susceptibility to cold treatments in a standardized series of exposures and temperatures. These results are expected to further encourage the “real world” application of cold treatments for the disinfestation of durable stored-products.     

Genetic diversity and population structure analysis of <Emphasis Type="Italic">Pistacia</Emphasis> species revealed by <Emphasis Type="Italic">phenylalanine ammonia</Emphasis>-<Emphasis Type="Italic">lyase</Emphasis> gene markers and implications for conservation

Information on population genetic structure and crop genetic diversity is important for genetically improving crop species and conserving threatened species. The PAL gene sequence is part of a multigene family that can be utilized to design DNA marker systems for genetic diversity and population structure investigation. In the current study, genetic diversity and population structure of 100 accessions of wild Pistacia species were investigated with 78 PAL markers. A protocol for using PAL sequences as DNA markers was developed. A total of 313 PAL loci were recognized, showing 100% polymorphism for PAL markers. The PAL markers produced relatively more observed and effective alleles in Pistacia falcata and Pistacia atlantica, with a higher Shannon’s information index and expected heterozygosity in P. atlantica, Pistacia vera and Pistacia mutica. Pairwise assessment of Nei’s genetic distance and genetic identity between populations revealed a close association between geographically isolated populations of Pistacia khinjuk and Pistacia chinensis. The accessions of wild Pistacia species had more genetic relationship among studied groups of species. Analysis of molecular variance indicated 19% among-population variation and 81% within-population variation for the PAL gene based DNA marker. Population structure analysis based on PAL revealed four groups with high genetic admixture among populations. The results establish PAL markers as a functional DNA marker system and provide important genetic information about accessions from wild populations of Pistacia species.