Diagnostic molecular markers of six lepidopteran insect pests infesting apples in Korea

Two molecular identification techniques for differentiating six lepidopteran pests infesting apples in Korea are presented. These six species include two internal fruit feeders (Grapholita molesta and Carposina sasakii), two leaf rollers (Adoxophyes sp. and Archips breviplicanus) and two leaf miners (Phyllonorycter ringoniella and Lyonetia prunifoliella). All species occur until near harvest and reduce apple production. A 489 bp fragment of mitochondrial cytochrome oxidase subunit I (COI) was sequenced in these six species. The sequence was used to select species-specific restriction enzyme sites and to design diagnostic polymerase chain reaction (PCR) primers, resulting in the development of restriction fragment length polymorphism (RFLP)-PCR and diagnostic PCR. These methods were reliable and rapid in the identification of these six species.     

The present distribution and predicted geographic expansion of the floodwater mosquito Aedes sticticus in Sweden

The mass emergence of floodwater mosquitoes, in particular Aedes sticticus and Aedes vexans, causes substantial nuisance and reduces life quality for inhabitants of infested areas and can have a negative impact on the socio‐economic conditions of a region. We compared the previous, present, and predicted geographic distribution of Ae. sticticus in Sweden. Previous records from the literature until 1990 list the species in three out of 21 Swedish counties. Beginning in 1998, studies show that the present distribution of the species covers 11 counties, with highest abundances in an east‐west belt in Central Sweden. Using climate data from the present and predicted climate scenarios, the expected distribution of Ae. sticticus in 2020, 2050, and 2080 could be modelled using GIS. As variables, mean temperatures and cumulative precipitation between May and August and degree slope were chosen. The predicted geographic distribution of Ae. sticticus will continue to increase and include 20 out of 21 Swedish counties. The expected temperature rise will increase the suitable area towards the northern part of Sweden by 2050. Some non‐suitable areas can be found along the south‐east coast due to insufficient amount of precipitation in 2050 and 2080. Modelling the expected distribution of a species using predicted climate change scenarios provides a valuable tool for risk assessments and early‐warning systems that is easily applied to different species and scenarios.     

A diagnostic multiplex polymerase chain reaction method to identify Japanese internal apple-feeding Lepidopteran pests: Grapholita molesta, Grapholita dimorpha (Lepidoptera: Tortricidae), and Carposina sasakii (Lepidoptera: Carposinidae)

We present a multiplex polymerase chain reaction method to differentiate between three Japanese internal apple-feeding pests: the oriental fruit moth, Grapholita molesta (Busck), Grapholita dimorpha Komai, and the peach fruit moth, Carposina sasakii Matsumura. A 1,342-bp fragment of mitochondrial cytochrome oxidase subunit I (COI) was sequenced in each species. The three species showed consistent and diagnostic differences in the region of the COI gene, from which three species-specific forward primers were designed. The forward primers along with one universal reverse primer were used to selectively amplify DNA from specimens of diverse geographic origin for each corresponding target species. This method enabled easy, immediate, and accurate identification of internal feeding Lepidoptera in apples and other fruits.     

Laboratory rearing of Lytopylus rufipes (Hymenoptera: Braconidae: Agathidinae), a parasitoid wasp of the oriental fruit moth,Grapholita molesta (Lepidoptera: Tortricidae), using apple and a commercially available diet

Lytopylus rufipes (Nees) (Hymenoptera: Braconidae: Agathidinae) is a potential natural enemy of the oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), but there is no established method to rear this wasp continuously. In the laboratory, female wasps can produce both female and male progenies without mating (deuterotokous), but host-infested plants are necessary to trigger oviposition behavior. In this study, immature apples were used because they keep well. Grapholita molesta larvae were transferred to immature apples, and then exposed to L. rufipes females. After parasitization, these apples were transferred to blocks of artificial diet (Silkmate 2M) for further rearing. Using this transitional diet system, L. rufipes females develop in 25.1?±?1.8 (mean?±?SD) days from egg to adult, but male wasps require only 23.8?±?1.0 days. Furthermore, the longevity of female wasps was 12.2?±?7.3 (mean?±?SD) days, and the parasitism rate was 26.4 (95% Cl: 22.2–30.6). Female wasps can attack host larvae in all instars, but parasitism of first-instar larvae is more successful. Therefore, first-instar larvae of G. molesta are recommended for establishing a L. rufipes colony under laboratory conditions. This system decreases the requirement for plant material and maintains continuous production of L. rufipes.

     

A novel approach to the assessment of fruit damage caused by <Emphasis Type="Italic">Carposina sasakii</Emphasis> (Lepidoptera: Carposinidae) using pheromone trap catches in Korean orchards

This study reports a model that utilizes pheromone trap catches to assess the fruit damage caused by Carposina sasakii. The model consisted of four steps: (1) obtaining influx population density using pheromone traps, (2) estimating the actual female population within a defined area using an estimated conversion rate, (3) calculating the total number of eggs using the oviposition model of C. sasakii, and (4) estimating the proportion of fruits infested with eggs (potential damaged fruits) using the relationship between mean egg density per fruit and the proportion of fruits infested with eggs. The relationship between mean egg density ([`(x)] \bar{x} ) per fruit and variance (s ²) was well described by Taylor’s power law, and its parameters were successfully incorporated into the equation that estimates the relationship between mean egg density and the proportion of fruits infested with eggs. In peach orchards, the model accurately predicted the proportion of fruits infested with eggs at the beginning of C. sasakii emergence in early season, but overestimated it in the mid-season. The fitting ability of the model outputs largely increased when the factor of oviposition behavior of C. sasakii was incorporated into the simulation processes, applying the allocation module of total eggs between peaches and apples.     

Different types of fruit damages of three internal apple feeders diagnosed with mitochondrial molecular markers

Three tortricid pests, Grapholita dimorpha (Komai), G. molesta (Busck), and Carposina sasakii (Matsumura), are known as internal apple feeders in Korea. To identify young larvae, this study developed two types of molecular markers from their mitochondrial DNA (mtDNA) sequences. To this end, six different loci of mtDNA were sequenced in G. dimorpha: cytochrome oxidase subunit I (460 bp), cytochrome oxidase subunit II (446 bp), cytochrome b (308 bp), NADH dehydrogenase 3 (585 bp), NADH dehydrogenase 4 (ND4, 835 bp), and 16S rRNA (1300 bp). These sequences were compared with those of G. molesta and C. sasakii in order to develop PCR–RFLP and diagnostic primers. ND4 locus was selected to be used for developing a PCR–RFLP marker. ND4-Swa I digests showed two bands for G. dimorpha, one band for G. molesta, and three bands for C. sasakii. On the other hand, species-specific diagnostic PCR primers were developed using ND4 locus. These markers were then applied to diagnose larvae infesting apples to determine species-specific fruit damage patterns, in which G. dimorpha, G. molesta, and C. sasakii showed different feeding behaviors in terms of their main feeding sites in apple fruits.     

Field observations quantifying attraction of four tortricid moths to high-dosage pheromone dispensers in untreated and pheromone-treated orchards

Orientational responses of four species of feral tortricid moths (Lepidoptera: Tortricidae) to polyethylene tube dispensers of pheromone were observed in a 0.8 ha apple orchard treated with such pheromone dispensers and in an untreated 0.8 ha orchard. Male oblique‐banded leafrollers, Choristoneura rosaceana (Walker) (mean 7.2 ± 0.4 moths/night during 21 nights), Oriental fruit moths, Grapholita molesta (Busck) (mean 10.5 ± 2.1 during 20 evenings), and the redbanded leafrollers, Argyrotaenia velutinana (Walker) (mean 2.0 ± 1.1 during 14 nights) were observed approaching within 100 cm of their respective polyethylene‐tube pheromone dispensers in the untreated orchard. Furthermore, C. rosaceana (mean 2.0 ± 0.7 during 17 nights) and G. molesta (mean 1.5 ± 0.4 over 20 evenings) came within 100 cm of their respective polyethylene‐tube pheromone dispensers in the pheromone‐treated orchard. Most visits lasted less than 10 s, after which the majority of moths departed by flying upwind. In the untreated orchard, the number of C. rosaceana observed orienting to polyethylene tube dispensers was greater than the number captured in optimized monitoring traps (1.9 ± 0.4) per night of observation. The numbers of A. velutinana (2.0 ± 1.1) or G. molesta (10.5 ± 2.1) attracted to polyethylene‐tube dispensers in the untreated orchard did not differ statistically from the numbers captured in optimized monitoring traps per night of observation. In the pheromone‐treated orchard, the number of C. rosaceana (2.0 ± 0.4) or G. molesta (1.2 ± 0.2) observed orienting to polyethylene‐tube dispensers did not differ statistically from the numbers of male moths of these species captured in optimized monitoring traps per night of observation. No codling moths, Cydia pomonella L. were observed orienting to, or landing near, their respective polyethylene‐tube dispensers in either the untreated or pheromone‐treated orchards, although substantial numbers were captured in monitoring traps per night of observation (6.0 ± 1.7) in the untreated orchard. The attraction of male moths to polyethylene tube dispensers thus occurred in three of the four species observed. These results provide support for the idea that false‐plume‐following is an important component of the mechanisms mediating communicational disruption in moths by polyethylene‐tube dispensers.     

Model of Grapholita molesta spring emergence in pear orchards based on statistical information criteria

The oriental fruit moth, Grapholita molesta, is becoming a large threat to Korean pear production. Timely management of the egg and early larval stages from the spring emergence is critical to reduce the G. molesta population during the pear growing season. A model was developed to precisely predict the spring occurrence of G. molesta adults as a function of accumulated degree-days. The model was validated with male moth caught in sex pheromone-baited traps placed in pear orchards at two major pear production regions (Icheon and Naju) of Korea in 2010. We applied nine distribution models to describe the cumulative proportions of G. molesta males caught relative to accumulated degree-days. The observed phenology of the G. molesta spring population was well described by the nine models. The predicted dates for the cumulative 50% male moth catches were within a 5 day period. Based on statistical information criteria (Akaike's and Bayes–Schwartz information criteria), we recommend the sigmoid function referred by Brown and Mayer, because of its ease of use and meaningfulness; the parameter “b” denotes the degree-day accumulation at 50% moth emergence. The G. molesta spring emergence model could be applied to determine optimal chemical treatment timing for controlling G. molesta in fruit tree orchards and further help to develop a full-cycle phenology model of G. molesta.     

Decontamination of coal mine effluent generated at the Rajrappa coal mine using phytoremediation technology

Toxicity of the effluent generated at the Rajrappa coal mine complex under the Central Coalfields Limited (CCL, a subsidiary of Coal India Limited) in Jharkhand, India was investigated. The concentrations (mg L^?1) of all the toxic metals (Fe, Mn, Ni, Zn, Cu, Pb, Cr, and Cd) in the coal mine effluent were above the safe limit suggested by the Environmental Protection Agency (EPA 2003). Among these, Fe showed the highest concentration (18.21 ± 3.865), while Cr had the lowest effluent concentration (0.15 ± 0.014). Efforts were also made to detoxify the effluent using two species of aquatic macrophytes namely “‘Salvinia molesta and Pistia stratiotes.” After 10 days of phytoremediation, S. molesta removed Pb (96.96%) > Ni (97.01%) > Cu (96.77%) > Zn (96.38%) > Mn (96.22%) > Fe (94.12%) > Cr (92.85%) > Cd (80.99%), and P. stratiotes removed Pb (96.21%) > Fe (94.34%) > Ni (92.53%) > Mn (85.24%) > Zn (79.51%) > Cr (78.57%) > Cu (74.19%) > Cd (72.72%). The impact of coal mine exposure on chlorophyll content showed a significant decrease of 42.49% and 24.54% from control values in S. molesta and P. stratiotes, respectively, perhaps due to the damage inflicted by the toxic metals, leading to the decay of plant tissues.     

Projections of global and UK bioenergy potential from Miscanthus × giganteus—Feedstock yield,carbon cycling and electricity generation in the 21st century

In this article, we modify bioenergy model MiscanFor investigating global and UK potentials for Miscanthus × giganteus as a bioenergy resource for carbon capture in the 21st century under the RCP 2.6 climate scenario using SSP2 land use projections. UK bioenergy land projections begin in the 2040s, 60 year average is 0.47 Mega ha rising to 1.9 Mega ha (2090s). Our projections estimate UK energy generation of 0.09 EJ/year (60 year average) and 0.37 EJ/year (2090s), under stable miscanthus yields of 12 t ha^?1 year^?1. We estimate aggregated UK soil carbon (C) increases of 0.09 Mt C/year (60 year average) and 0.14 Mt C/year (2090s) with C capture plus sequestration rate of 2.8 Mt C/year (60 year average) and 10.49 Mt C/year (2090s). Global bioenergy land use begins in 2010, 90 year average is 0.13 Gha rising to 0.19 Gha by the 2090s, miscanthus projections give a 90 year average energy generation of 16 EJ/year, rising to 26.7 EJ/year by the 2090s. The largest national capabilities for yield, energy and C increase are projected to be Brazil and China. Ninety year average global miscanthus yield of 1 Gt/year will be 1.7 Gt/year by the 2090s. Global soil C sequestration increases less with time, from a century average of 73.6 Mt C/year to 42.9 Mt C/year by the 2090s with C capture plus sequestration rate of 0.54 Gt C/year (60 year average) and 0.81 Gt C/year (2090s). M. giganteus could provide just over 5% of the bioenergy requirement by the 2090s to satisfy the RCP 2.6 SSP2 climate scenario. The choice of global land use data introduces a potential source of error. In reality, multiple bioenergy sources will be used, best suited to local conditions, but results highlight global requirements for development in bioenergy crops, infrastructure and support.     

Life history of the Asian longhorn beetle Anoplophora glabripennis (Coleoptera Cerambycidae) in southern Europe

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Control effect on the brown-marmorated stink bug, <Emphasis Type="Italic">Halyomorpha halys</Emphasis> (Hemiptera: Pentatomidae), by combined spraying of pyrethroid and neonicotinoid insecticides in apple orchards in northern Japan

Control of Halyomorpha halys (Stål) adults by combined spraying of pyrethroid and neonicotinoid insecticides has been investigated in apple orchards in northern Japan in 2008 and 2009. Insecticides were selected to control not only H. halys but also the common lepidopteran pests Carposina sasakii Matsumura, Adoxophyes orana fasciata Walsingham, and Phyllonorycter ringoniella Matsumura. Dinotefuran was sprayed in mid-May and early July, fenpropathrin in late June, and bifenthrin in late July. H. halys adults were released into apple trees at 3–5-day intervals from mid-May to mid-August. In this spraying program, survival of the adults increased 6 days after spraying, but relatively few injury marks were noted throughout the growing season. On the other hand, in the conventional spraying program, the survival ratio of the adults increased rapidly 3 days after spraying—especially after the organophosphate applied in mid-May, late June, and late July—and increases in the number of injury marks synchronized well with survival in both years. This spray program reduced the number of fruit injury marks to one-fifth that of the conventional spraying program and one-tenth that of fungicides alone.     

Die küstenvegetation ostkanadas: D. Thannheiser. Münstersche Geographische Arbeiten. No. 10. Schöningh,Paderborn, 1981, 201 pp. 166 figs., 41 tables,DM. 41.50. ISBN 3-506-73210-2

Temperatures 10 and 2 cm above water level, in leaves of Salvinia molesta Mitchell, and 2 and 10 cm below water level, were measured in the field at hourly intervals over a total of 125 days. Temperature cycles of leaves and air had mean diurnal amplitudes of 8°C in summer and 17°C in winter; cycles in water lagged behind and had amplitudes which decreased with depth. Most parts of S. molesta were warmer than the air at a nearby weather station most of the time and there was temperature stratification in the water during the warm part of each day.Standard meteorological variables were selected, using stepwise regression, to predict daily maximum and minimum temperatures of S. molesta. Thermal inertia of water in the lake seemed to elevate S. molesta temperatures in autumn and depress them in spring compared with temperatures at the weather station. Better predictors were obtained by adding to meteorological variables a function based on the annual cycle of temperatures 10 cm below water level. Hourly temperatures experienced by S. molesta were predicted using curves fitted to diurnal cycles. The reliability of predictions was tested for each season of the year with independent data for a total of 78 days. Predicted temperatures were close to observed temperatures both in absolute terms and in terms of temperature-dependent growth rates of S. molesta.     

Emergence and mating behavior of the oriental fruit moth Cydia molesta (Lepidoptera: Tortricidae) and its potential for reproduction

The Oriental fruit moth, Cydia molesta (Busck, 1916) (Lepidoptera: Tortricidae), is a key pest of fruit and is widely distributed around the world. There are important connections between its behavior and biology and its management in agriculture, but few studies have investigated the associations between adult behaviors and oviposition. In this study, adult emergence, mating, and reproduction were investigated under laboratory and field conditions. The ratio of females to males at eclosion was approximately 1:1. When one virgin female had access to one virgin male, 66% and 34% of the couples copulated just once and twice, respectively; and the infertility rate of eggs (21.39 ± 1.25%) did not vary daily. Males, given access to one new female daily, could copulate multiple times, whereas females seldom mated more than once, indicating a male-biased operational sex ratio, but mating status of the male parent had no effect on progeny egg reproduction. Also, the number of eggs that hatched by all female partners of a male was inversely proportional to copulation duration for the female laying the eggs for total female reproductive success; and the number of eggs laid by all female partners of a male was proportional to their number of matings for total male reproductive success. However, the total number of eggs that hatched did not significantly differ for eggs laid by a female given new virgin males daily for mating (17.75 ± 4.28) versus eggs laid by virgin females (19.17 ± 7.51) presented daily with a male that re-mated daily with the series of females. Therefore, our results showed that females engaged in mate choice and males engaged in mate competition, affecting egg production, a factor that may be used to enhance mating disruption technology against Cydia molesta.     

Temperature-related effects of three insects on growth ofSalvinia molesta in Brazil

The effect ofCyrtobagous salviniae, Samea multiplicalis, andPaulinia acuminata on the growth ofSalvinia molesta was assessed in relation to temperature in field cages at 2 sites in Brazil, a canal with running-water and a lagoon with stillwater. Each insect species reduced the growth ofS. molesta over a mean leaftemperature range of 16–30°C, causing more damage as temperature increased. There was no difference between the 3 species in response to temperature. These results and their implication to the performance of these insects as biological control agents againstS. molesta are discussed.

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Résumé L'effet deCyrtobagous salviniae, deSamea multiplicalis et dePaulinia acuminata sur la croissance deSalvinia molesta a été étudié en relation avec la température dans des cages implantées dans 2 situations au Brésil, un canal avec de l'eau courante et une lagune avec de l'eau stagnante. Chaque espèce réduit la croissance deS. molesta dans une gamme de températures des feuilles s'étendant de 16° à 30°C, les dégats augmentent avec l'accroissement de la température. Il n'y a pas de différence entre les 3 espèces vis-à-vis de leur réaction à la température. Ces résultats et leur implication quant au rendement de ces insectes comme agents biologiques de lutte contreS. molesta sont discutés.

     

Molecular identification of glutathione S-transferase genes and their potential roles in insecticides susceptibility of Grapholita molesta

Grapholita molesta is one kind of serious fruit-boring pests in the world, which caused huge economic losses to the fruit industry including pear and apple. The heavy usage of synthetic insecticides resulted in different degrees of resistance of G. molesta to some commonly used insecticides. However, the tolerance mechanisms of G. molesta to insecticides are still unclear. Glutathione S-transferases (GSTs) belong to the superfamily of detoxifying enzymes and mainly play important roles in detoxification metabolism and insecticide susceptibility of insects. In this study, we first evaluated the toxicity of three insecticides including emamectin benzoate, chlorantraniliprole and lambda-cyhalothrin on G. molesta at different developmental stages. Synergism study showed that the susceptibility of G. molesta larvae to these three insecticides obviously increased when the enzyme activity of GST was inhibited by diethyl maleate compared with the controls. Next, we screened out six GmGST genes which are upregulated after insecticides treatment from a total of 21 GmGST genes and analysed their sequence characteristics and expression profiles. RNAi and bioassay results further revealed that the mortality of 5th instar larvae significantly increased after exposure to LC₃₀ of emamectin benzoate, chlorantraniliprole and lambda-cyhalothrin when silencing of GmGSTo2, GmGSTs1 and GmGSTz1 compared with the controls. In conclusion, this study indicated that GmGSTo2, GmGSTs1 and GmGSTz1 play important roles in insecticides susceptibility of G. molesta to three commonly used insecticides, which is significance for its future integrated management.     

Studies on decomposition of two species of Salvina

Decomposition of two species of Salvina, S. molesta Mitchell and S. cucullata Roxb. was studied for 220 days by the litter bag method. Salvinia molesta decomposes faster in the initial stages and S. cucullata in the later stages of decomposition. The release of nutrients is rapid during the first seven days and is ascribed to leaching. The rate of release of nutrients differs in the two species, with higher rates for P, Ca and Mg in S. molesta and for N in S. cucullata.     

High‐resolution prediction of leaf onset date in Japan in the 21st century under the IPCC A1B scenario

Reports indicate that leaf onset (leaf flush) of deciduous trees in cool‐temperate ecosystems is occurring earlier in the spring in response to global warming. In this study, we created two types of phenology models, one driven only by warmth (spring warming [SW] model) and another driven by both warmth and winter chilling (parallel chill [PC] model), to predict such phenomena in the Japanese Islands at high spatial resolution (500 m). We calibrated these models using leaf onset dates derived from satellite data (Terra/MODIS) and in situ temperature data derived from a dense network of ground stations Automated Meteorological Data Acquisition System. We ran the model using future climate predictions created by the Japanese Meteorological Agency's MRI‐AGCM3.1S model. In comparison to the first decade of the 2000s, our results predict that the date of leaf onset in the 2030s will advance by an average of 12 days under the SW model and 7 days under the PC model throughout the study area. The date of onset in the 2090s will advance by 26 days under the SW model and by 15 days under the PC model. The greatest impact will occur on Hokkaido (the northernmost island) and in the central mountains.     

Climate change affects the early-life history of a freshwater turtle in a severely drying region

Freshwater turtles are one of the most threatened vertebrate groups. Climate change is a major threat to these species, with impacts affecting all life-history stages. There is currently a limited understanding of how changes in climate may alter the environmental triggers for hatching and emergence from the nests of freshwater turtle hatchlings. This precludes making predictions about how climate change may impact freshwater turtle recruitment success. The southwestern snake-necked turtle (Chelodina oblonga) is endemic to south-western Australia, a global biodiversity hotspot that has undergone severe climatic drying. Recruitment failure is thought to be occurring in many populations of the species. However, there is little understanding as to how environmental change may be influencing recruitment. This study aimed to: (1) determine the incubation duration and hatching and hatchling emergence success of C. oblonga, (2) determine if the species exhibits hatching or emergence synchrony and/or delayed emergence and (3) quantify the effects of temperature and rainfall on hatchling emergence. Using this information, the study assesses how climatic drying and warming may be impacting C. oblonga's early life-history. Between 2018 and 2020 nest sites were monitored around a large urban wetland with weekly assessments of egg and hatchling status. Incubation duration and hatching and hatchling emergence success were calculated, and generalized linear models were built to determine how temperature and/or rainfall predicted emergence. Hatchlings either emerged shortly after hatching or overwintered in the nest, and both hatching and emergence were asynchronous. Both emergence periods were positively associated with temperature and rainfall. This study reveals that incubation duration, hatching success, hatchling emergence and survival are all likely to be impacted by recent and projected climate change, and especially drying. Warming and drying are predicted for many temperate regions globally, and it is therefore important that their impacts on the early life history of freshwater turtles be better understood.