Important Quarantine Pest Research Articles

Volatiles potentially useful to be used as biomarkers for monitoring the khapra beetle Trogoderma granarium for quarantine

The khapra beetle, Trogoderma granarium, is one of the most important quarantine pests globally. In order to identify potential biomarkers for establishing a rapid and effective monitoring technology for T. granarium quarantine, headspace-solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrum (GC-MS) was used to identify volatiles that are potentially useful for early detection of the presence of the T. granarium presence. Three volatiles, including Ethyl palmitate, (Z)-5-tetradecenoic acid and (Z)-14-methyl-8-hexadecen-1-al, were commonly identified as unique volatiles emitted from four different stored products including peanut, barley, wheat and rapeseed only after T. granarium infestation. In addition, four volatiles including hexadecane, palmitic acid, (Z)-14-methyl-8-hexadecen-1-al and N-heptadecane, were found emitted commonly from the larvae of T. granarium after they fed on the four stored products. These common chemicals could potentially be biomarkers sensitive enough for monitoring the presence of T. granarium. In addition to these unique chemicals, a combination of up- and down-regulated volatiles by each type of stored seeds were also identified as potential markers useful for the early detection of the presence of T. granarium. Our results could be a first step towards establishing a method for T. granarium monitoring and quarantine.

Biology of Citrus long-horned beetle, Anoplophora chinensis (Forster, 1771) (Coleoptera: Cerambycidae) on hazelnut

Anoplophora chinensis (Forster, 1771) (Coleoptera: Cerambycidae) is an important quarantine pest causing significant economic losses. It was detected in hazelnut orchards in Trabzon province in 2017 and an eradication program continues. In this study, the population level and biology of the pest were determined by rearing it and conducting periodic surveys in hazelnut orchards in Trabzon province Maçka district between 2020 and 2021. It has been revealed that the adult emergence of A. chinensis takes place in the last weeks of June and adults are found in nature until the end of August. The oviposition period continued from the end of June to the beginning of September and the larval stage of A. chinensis was observed throughout the year. The pupal stage was observed in May and June. With this study, the biology of A. chinensis in Türkiye has been revealed in detail for the first time and obtained data guides the eradication and surveillance studies.

Comparison of different attract-and-kill device densities to control the adult population of Popillia japonica (Coleoptera: Scarabaeidae).

Popillia japonica Newman is a scarab beetle native to Japan that is considered a serious pest outside its native range. It can feed on more than 400 host plants and spread about 10 km per year in invaded territories, therefore it is considered the second most important quarantine pest in Europe. Both chemical and biological insecticides have been used for control, with variable results. Despite ongoing efforts, P. japonica remains a threat in invaded countries, and it is necessary to test more effective and sustainable solutions in the context of integrated pest management. Here we present a study on long-lasting insecticide-treated nets (LLINs) assembled in semiochemical-baited attract-and-kill devices (A&Ks) as a means to control adults of P. japonica with low environmental impact. This study complements previous ones in which we first evaluated the effectiveness of the LLINs in the laboratory and then tested both effectiveness and duration in field-exposed A&Ks against P. japonica. In the present work we compared the effectiveness of three different densities of A&Ks per hectare in areas where the population of P. japonica was numerically homogeneous. The different densities of A&K (1, 2, 4 A&Ks per ha) resulted in an overall reduction of the population of P. japonica by about two thirds in comparison to the control area. This study suggests that the use of one A&K per hectare, requiring minimal management effort, is an effective ratio for reducing local populations of P. japonica. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Consumption of β-Caryophyllene Increases the Mating Success of Bactrocera zonata Males (Diptera: Tephritidae).

The peach fruit fly, Bactrocera zonata (Saunders) (Diptera: Tephritidae), is an economically important polyphagous quarantine pest of horticultural crops endemic to South and Southeast Asia. Methyl eugenol (ME), a naturally occurring phenylpropanoid, is a male attractant used to lure and (when mixed with an insecticide) annihilate the males from the wild population, a method of pest control termed the male annihilation technique (MAT). ME is reported to enhance the mating success of sterile males of Bactrocera spp., which is critical for enhancing the effectiveness of the sterile insect technique (SIT). The suppressed response of ME-treated males to ME-baited traps/devices allows the simultaneous application of the MAT and SIT, increasing the efficiency of area-wide integrated pest management (AW-IPM) programs. However, ME treatment in sterile males in SIT facilities is logistically difficult. β-caryophyllene (BCP) is a widely occurring, safer plant compound and is considered suitable for treating males in SIT facilities. Here, we demonstrate that BCP feeding enhanced B. zonata male mating success to the same extent as ME feeding. Feeding on BCP suppressed the male's subsequent attraction to ME-baited traps, but not to the same degree as feeding on ME. The results are discussed and BCP is suggested as an alternative to ME for the concurrent use of the MAT and SIT.

Attractant potential of Enterobacter cloacae and its metabolites to Bactrocera dorsalis (Hendel).

Bactrocera dorsalis (Hendel) has a wide host range. It has been the most important quarantine pest in many countries or regions. Currently, chemical control and bait trapping are mainly used in the monitoring, prevention, and control of B. dorsalis. However, chemical control will cause pollution of the environment and drug resistance of insects. Methyl eugenol, the main attractant currently used, can only attract males of B. dorsalis. This study focused on the attractant function and active substances of one key intestinal bacterium, Enterobacter cloacae, which was isolated from B. dorsalis. First, the attraction of the E. cloacae autoclaved supernatant to male and female adults of 0, 6, and15days post-emergence was confirmed using a Y-type olfactometer. Subsequently, through metabolome sequencing and bioassays, L-prolinamide was identified and confirmed as the most effective attractant for B. dorsalis. Finally, the synergistic effect of L-prolinamide with the sex attractant ME was validated through field experiments. This study confirmed the attraction effect of E. cloacae on B. dorsalis and also proved the attraction effect of L-prolinamide, the metabolite of E. cloacae, on B. dorsalis. This laid a theoretical foundation for the development of a new attractant and safe, green, and efficient prevention and control technology of B. dorsalis.

The Effects of Nine Compounds on Aldehyde-Oxidase-Related Genes in Bactrocera dorsalis (Hendel).

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) (B. dorsalis) is an important agricultural, major invasive, and quarantine pest that can cause significant damage to the economic value of the fruit and vegetable industry. Male bait is one of the most effective methods of surveying, monitoring, and controlling B. dorsalis. In our study, we constructed cDNA libraries using total RNA extracted independently from the antennae, mouthparts, and thoracic legs of male and female adults and the ovipositors of female adults and screened out four aldehyde-oxidase-related genes (AOX-related), C58800, C66700, C67485, and C67698. Molecular docking predictions showed that eight compounds, including 3,4-dimethoxycinnamyl alcohol, 3,4-dimethoxy-cinnamaldehyde, deet, ethyl N-acetyl-N-butyl-β-alaninate, n-butyl butyrate, n-butyl butyrate, ethyl butyrate, methyl eugenol, and ethyl acetate, could combine with proteins encoded by the four B. dorsalis AOX-related genes. Furthermore, QPCR was performed to confirm that four compounds, including 3,4-dimethoxy cinnamic aldehyde, butyl levulinic acid ethyl ester (mosquito repellent), butyl butyrate, and methyl eugenol, induced significant changes in the AOX-related genes of B. dorsalis. These results provide useful information and guidance for the batch screening of potentially useful compounds and the search for effective attractants of B. dorsalis.

Virulence of entomopathogenic bacteria Serratia marcescens against the red palm weevil, Rhynchophorus ferrugineus (Olivier).

The red palm weevil (RPW), Rhynchophorus ferrugineus (Olivier), is an important quarantine pest, which has caused serious economic losses in various palm species, such as coconut, oil palm and date palm. Finding effective biocontrol resources is important for the control of this pest and the protection of palm crops. A pathogenic strain HJ-01 was isolated from infected and dead pupa of Tenebrio molitor using tissue separation method. The HJ-01 strain was streak cultured and purified, and its morphological, physiological, biochemical characteristics, and 16S rDNA homology were identified after conducting a pathogenicity test on RPW larvae. Strain HJ-01 exhibited remarkable pathogenicity against RPW larvae. Under the concentration of HJ-01 suspension was 1.0 × 108 cfu/mL, the mortality rate of RPW reached 82.22%, and the half-lethal time (LT50) was 4.72 days. RPW larvae infected with strain HJ-01 showed reduced movement, decreased appetite, and eventual death. As the treatment progresses, the larvae's bodies turned red, became soft, and started to rot, resulting in the discharge of liquid. HJ-01 demonstrated the ability to produce scarlet pigment after 24 h of culture on a basic medium. Colonies of HJ-01 appeared convex, bright red, moist, and viscous, opaque in the center, irregular at the edges, and emitted an unpleasant odor. Under microscopic observation, the cells of HJ-01 appeared as short rod-shaped and flagellate, with a size ranging from (1.2-1.8) μm × (1.0-1.2) μm. Genomic DNA extraction was performed on the strain, and the 16S rDNA sequence was amplified, yielding a sequence length of 1445 bp. The sequence of strain HJ-01 displayed a 99.72% similarity to that of Serratia marcescens. Phylogenetic tree analysis further confirmed that strain HJ-01 belonged to S. marcescens. Therefore, the strain HJ-01 has a very good lethal effect on RPW larvae, and it may be used as an effective bacterium for the control of RPW.

UDA-FlyRecog: Unsupervised domain adaptation for drosophila cross-domain recognition model

Drosophila is an important quarantine pest, and its identification plays a crucial role in agricultural development. To address the issue of significant style variations in Drosophila image samples collected from different environments, which results in poor performance when applying models trained on the source domain to the target domain, this paper proposes a domain-adaptive model for cross-domain Drosophila recognition (UDA-FlyRecog). This model combines global feature alignment and class-aware feature alignment methods, and adopts a cyclic iterative training approach to mitigate the effects of domain shift. Experimental results demonstrate that our method achieves the highest accuracy improvement of up to 16.7% compared to other methods on Drosophila datasets collected from both laboratory and natural environments, indicating promising practical applications.

New and rapid visual detection assay for Trogoderma granarium Everts based on recombinase polymerase amplification and CRISPR/Cas12a.

Khapra beetle (Trogoderma granarium Everts), one of the most important quarantine pests globally, is capable of causing severe infestation and huge economic loss to stored grain, and its interception rate has increased in major global trade countries over the past few years. However, difficulties remain in distinguishing this species with similar ones. In order to assist border ports and warehouses in khapra beetle's effective rapid identification as well as pest control at the early stages of monitoring or interception, we herein developed a new and rapid visual detection assay for T. granarium based on recombinase polymerase amplification (RPA) and CRISPR/Cas12a system. We designed and selected the first khapra beetle-specific RPA primers and crRNA, and optimized the visualization reaction system (Cas12a/CrRNA=100 nM/500 nM). With only a 37°C-heat-source and a blue light torch, RPA and CRISPR/CAS12a-based visualization assays can be completed within 40 min to differentiate among khapra beetle and other nine similar Dermestidae species. After DNA extraction using a kit (4-5 h) or a simple method (5 min), the specific amplicons were obtained after a 15 min-RPA-reaction at 37°C, followed by a 15 min-color-reaction under 37°C in dark using CRISPR/CAS12a system and a fluorescent probe (5'-FAM/3'-BHQ1 labeled). This method is ingenious to low levels of DNA (10-1 ng/μL) and meets the sensitivity requirements for detecting a single khapra beetle's egg (about 0.7 mm). Our specificity and sensitivity analysis inferred that the present visualization system is effective to quickly and uniquely detect khapra beetle at room temperature (37°C), thereby preventing this species before they spread widely. Our study is suitable for being pushed forward in storage pest management and provides value as a reference for monitoring and identification of other pests. This article is protected by copyright. All rights reserved.

Phytosanitary irradiation treatment of the aerial root mealybug, Pseudococcus baliteus (Hemiptera: Pseudococcidae).

The aerial root mealybug, Pseudococcus baliteus Lit (Hemiptera: Pseudococcidae), is an important invasive and quarantine pest that poses a potential threat to fruits, vegetables, and ornamental plants. As a result, phytosanitary treatments are necessary to ensure the commodities of international trade are free from these pests. To determine the minimum absorbed dose required for phytosanitary irradiation (PI) application, irradiation dose-response and large-scale confirmatory tests were conducted. Eggs that were 2, 4, and 6 days old and late gravid females (containing 0-day-old eggs) of P. baliteus were X-ray irradiated with doses of 10, 20, 40, 60, 80, 100, and 120 Gray (Gy). The efficacy of preventing egg-hatching (mortality) was compared using two-way ANOVA, 95% confidence interval overlapping and lethal dose ratio test in probit analysis. The radiotolerance sequence of mealybugs egg was found to be 0 < 2 ≈ 4 < 6-day-old eggs, and their estimated LD99.9968 values with 95% confidence interval were 132.0 (118.9-149.5), 137.6 (125.2-153.7), 145.5 (134.5-159.1), and 157.4 (144.6-173.6) Gy, respectively. Subsequently, target doses of 135 and 145 Gy were used in the confirmatory gamma radiation treatments. No F1 generation neonates developed from a total of 47,316 late females irradiated at the measured dose of 107.7-182.5 Gy, resulting in the treatment efficiency of 99.9937% at the 95% confidence level. Therefore, the highest dose of 183 Gy measured in the confirmatory tests is recommended as the minimum absorbed dose in PI treatment of P. baliteus for establishing national and international standards.

Is there hybridization between 2 species of the same genus in sympatry?-The genetic relationships between Anoplophora glabripennis, Anoplophora chinensis, and putative hybrids.

Anoplophora glabripennis (Asian longhorn beetle, ALB) and Anoplophora chinensis (Citrus longhorn beetle, CLB) are native forest pests in China; they have become important international quarantine pests. They are found using the same Salix aureo-pendula host tree of Cixi, Zhejiang province, China. On this host tree, we collected additional beetles that appeared to be morphologically intermediate between ALB and CLB. By using a stereoscope, we observed that there were several bumps on the base of the elytra, which was inconsistent with ALB, which typically has a smooth elytral base, but was more like CLB, which has numerous short tubercles on the elytral base. Given their sympatry and intermediate morphology, we hypothesized that these may represent ALB×CLB hybrids. We studied the genomic profiles for 46 samples (ALB, CLB, and putative hybrids) using genotyping-by-sequencing (GBS) providing a reduced representation of the entire genome. Employing principal component analyses on the 163 GBS-derived single nucleotide polymorphism data, we found putative hybrids tightly clustered with ALB, but genetically distinct from the CLB individuals. Therefore, our initial hybrid hypothesis was not supported by genomic data. Further, while mating experiments between adult ALB and CLB were successful in 4 separate years (2017, 2018, 2020, and 2021), and oviposition behavior was observed, no progeny was produced. Having employed population genomic analysis and biological hybridization experiments, we conclude that the putative hybrids represent newly discovered morphological variants within ALB. Our approach further confirmed the advantage of genome-wide information for Anoplophora species assignment in certain ambiguous classification cases.

Prediction of the Current and Future Distribution of Tomato Leafminer in China Using the MaxEnt Model.

Tomato leafminer (Tuta absoluta), an important quarantine pest in China, was first detected in China in Yili, Xinjiang Uygur Autonomous Region, in 2017. Its damage has grown in recent years, severely harming Solanaceae plants in China and causing enormous economic losses. The study and prediction of the current and future suitable habitats for tomato leafminer in China can provide an important reference for the monitoring, early warning, and prevention and control of the pest. Here, tomato leafminer's potential distributions in China under the current climate and four future climate models (SSP1-26, SSP2-45, SSP3-70, and SSP5-85) were predicted using the maximum entropy (MaxEnt) model with ArcGIS software, and the accuracy of the prediction results was tested. The areas under the receiver operating characteristic curves of the models were all greater than 0.8, and the test omission rate of the model simulation results basically agreed with the theoretical omission rate, suggesting that the prediction results had satisfactory accuracy and reliability. Under the current climatic conditions, the highly suitable habitats for tomato leafminer in China are mainly distributed in most of North China, most of East China, most of South China, most of Central China, most of Southwest China, some parts of Northeast China, and only a few parts of Northwest China. Annual mean temperature is the main environmental factor limiting the distribution. The suitable habitats for tomato leafminer will shift under different future climate models: Under SSP1-26, the highly suitable habitats will spread to the north and northeast and to the southeast coastal areas; under SSP2-45, the size of highly suitable habitats will grow from the present to 2080 and shrink from 2081 to 2100; under SSP3-70, the highly suitable habitats will spread northeastwards, but the highly suitable habitats in southeast coastal areas will shrink from 2081 to 2100 and turn into moderately suitable habitats. Under SSP5-85, the highly suitable habitats will spread northeastwards and northwestwards, with the size of highly suitable habitats gradually decreasing and the size of moderately suitable habitats increasing. Different climates will lead to different distributions of suitable habitats for tomato leafminer, with annual mean temperature, isothermality, and mean diurnal range as the main environmental influences.

Lethal and sublethal effects of broflanilide on four tephritid pests (Diptera: Tephritidae).

Fruit flies are internationally important quarantine or invasive pests of many fruits and vegetables and can cause serious economic losses. Long-term reliance on insecticides for controlling these pests has led to increasing resistance to multiple insecticides; hence, a new agent is needed. In this study, the acute toxicity and sublethal effects of the novel insecticide broflanilide on four adult fruit fly species, Bactrocera dorsalis, Bactrocera cucurbitae, Bactrocera tau, and Bactrocera correcta, were evaluated. Broflanilide was effective against B. dorsalis and B. correcta, with lethal concentration values (amount required to kill 50% of animals; LC50 ) of 0.390 and 1.716 mg/L. However, for B. cucurbitae (19.673 mg/L) and B. tau (24.373 mg/L), the LC50 was 50-60 times higher than that of B. dorsalis. The survival rates of B. correcta and B. cucurbitae were significantly lower under LC50 treatment than those of the control (corrected for mortality rate). Sublethal concentrations of broflanilide stimulated fecundity in all species except B. tau. The hatching rate at LC50 was significantly lower for B. correcta and B. tau compared with the control and even more so for B. correcta, which was zero. Broflanilide is potentially an effective insecticide for controlling B. dorsalis and B. correcta. However, the variation in toxicity of broflanilide to the four fruit flies suggests that species variation needs to be carefully considered. Our results highlight the importance of clarifying the sublethal effects of insecticides on target insects to ensure the comprehensive evaluation and rational use of insecticides. © 2023 Society of Chemical Industry.

Effect of Chilling Temperature on Survival and Post-Diapause Development of Korean Population of Lymantria dispar asiatica (Lepidoptera: Erebidae) Eggs

One of the subspecies of the Eurasian spongy moth, Lymantria dispar asiatica, is a destructive forest pest in native regions and also an important quarantine pest in non-native regions. Its polyphagous nature, together with occasional outbreaks, may seriously threaten ecosystems and result in costly management programs. In this study, we examined the effect of chilling temperatures (−12, −6, 0, 6, and 12 °C) during the diapause phase on the survival and post-diapause development of L. d. asiatica eggs, collected before winter, in order to characterize their thermal response. The eggs were exposed to treatment temperatures for 100 days, followed by 25 °C incubation to determine their survival and development time. The eggs hatched in all the treatments, indicating that all the examined conditions could partly or sufficiently satisfy the thermal requirement for eggs to enter post-diapause development. However, exposure to chilling temperatures significantly affected both the survival and development times of overwintering eggs in a given temperature range. The survival rates declined at −12 °C, and the development rates accelerated as the chilling temperature increased. This information could offer clues for the assessment of the outbreak potential in native regions and the possibility of range expansion in non-native regions through the consideration of winter conditions that favor L. d. asiatica egg hatching and their subsequent development.

Gamma and X-ray irradiation as a phytosanitary treatment against various stages of Planococcus lilacinus (Hemiptera: Pseudococcidae)

• Late-aged adult female of cacao mealybug is most tolerant to gamma and X-ray irradiation. • None F 1 generation 2nd instar nymphs emerged from 97,384 adult females irradiated at 126.1-163.0 Gy in confirmatory tests. • The treatment efficacy is not less than 99.9969% at the 95% confidence level. • A minimum of 163 Gy is suggested for phytosanitary irradiation treatment of cacao mealybug. The cacao mealybug, Planococcus lilacinus Cock, is an important quarantine pest. Infested commodity should be subject to appropriate phytosanitary treatment, while irradiation is recommended for the cacao mealybug. Radio-tolerance comparison tests were conducted on the crawler, nymphs, and adult females of P . lilacinus at the X-ray radiation doses of 40, 80, and 120 Gy, respectively. The results showed that irradiation had a strong effect on preventing of development and reproduction; the adult female stage was identified as the most tolerant. During the following dose-response tests, among young and late females X-ray-irradiation (20–100 Gy), the late females were most tolerant when preventing F 1 generation 2nd instars emergence was used as the evaluation criterion. Minimum absorbed dose and its 95% fiducial limits to provide probit 9 efficacy at 95% confidence level (100% mortality/inhibition in an estimated population of 93,616 individuals) were 131.5 Gy (122.5, 142.6 Gy) and 144.4 Gy (132.7, 159.4 Gy), estimating from the probit analysis on dose-mortality data of 1–30 and 1–10-day-old neonates laid by late females, respectively. In the large-scale confirmatory tests, a total of estimating 97,384 late females of P. lilacinus rearing on the pumpkins fruits were irradiated with gamma-ray at the target dose of 135 or 145 Gy (measured doses 126.1–163.0 Gy), which resulted in no F 1 generation 2nd nymphs developing during a 6-week post-treatment period. The treatment efficacy calculated is 99.9969% at the 95% confidence level. Therefore, a minimum absorbed dose of 163.0 Gy is recommended for phytosanitary treatment of P. lilacinus in infested commodity.

First detection of the potato cyst nematode (Globodera rostochiensis) in a major potato production region of China.

The potato cyst nematodes (PCN) Globodera rostochiensis and Globodera pallida are the very important quarantine nematode pests of potato [Stone 1973]. Both species cause serious potato tuber yield losses. These species are subjected to strict quarantine regulations in many countries (EPPO 2017). G. rostochiensis was detected in Sichuan and Yunnan province, China in 2022 (Jiang et al. 2022). A survey for cyst nematodes in potato fields was conducted in Guizhou Province from 2018 to 2020. A total of 200 samples, including roots and soil, were collected from 40 potato fields in Hezhang (N27 06.145, E104 39.153) and Weining (N26 50.541, E104 09.885) counties in Guizhou Province, China. The Cobb decanting and sieving method was used to isolate cysts and J2s from the soil samples (Southey 1986). The potato roots were stained with acid fusion to observe cyst development. Morphological and molecular analyses indicated that 27 (13.5%) of the samples contained G. rostochiensis. The cyst density ranged between 1-85 cysts per 100 cm3 of soil and a mean density was 15 cysts per 100 cm3 soil. The smoothly rounded cysts were in brown and golden color, and the terminal cone was absent and circumfenestrate. The key morphometrics of cysts (n=20) were 695 ± 26 (685-757) μm in length excluding neck and 690 ± 30 (668 to769) μm in width; the number of cuticular ridges between anus and vulval fenestra was 16.3 ±2.1 (14 to 18); fenestral length was 15.1 ± 2.1 μm (13.18 to 19.27); distance from anus to the edge of fenestra was 61.12 ± 8.9 (49.22 to76.27) μm; and Granek's ratio was 4.54 ± 0.8 (3.97-5.26). The key morphometrics of J2 (n = 20): 468.0 ± 20.1 (427 to - 521) μm in body length, 20.58 ± 0.7 (20.2 to 21.8) μm in stylet length, 43.9 ± 5.6 (40.3 to 53.9) μm in tail length, and 23.1 ± 1.8 (21.77 to 25.32) μm in hyaline region length. The cyst and J2 morphologies were consistent with those of G. rostochiensis (Subbotin et al. 2010, EPPO 2017). Genomic DNA was isolated from cysts (n=20). DNA extraction was performed in a volume of 20 μl containing 3 μl 10× PCR buffer, 3 μl Proteinase K (600 μg μl-1), 14 μl distilled water and a single cyst was added and ground in an ice bath as described by Ou et al. (2008). The internal transcribed spacer (ITS) regions were amplified using the universal primers: rDNA1 (5'-TTGATTACGTCCCTGCCCTTT-3') and rDNA2 (5'-TTTCACTCGCCGTTACTAAGG-3') (Fleming 1998 ), and the 28S rDNA-D2/D3 regions were amplified using the primers: D2A (5´-ACAAGTACCGTGAGGGAAAGTTG-3´) and D3B (5´-TCGGAAGGAACCAGCTACTA-3´) (Subbotin et al. 2006). After the brackets at the beginning and end of the sequences were closed-up, the ITS rDNA sequences (GenBank Accession No. MZ042367 and MZ042368) showed 99.66% - 99.92% identity to G. rostochiensis sequences available in GenBank (FJ212166.1, GQ294513, FJ212164.1 and KJ409617.1). Sequences from the 28S region (GenBank Accession No. MZ057597 and MZ057598) were 99.23% - 99.74% similar to those of G. rostochiensis isolate from Slovakia (KJ409625.1), Italy (KJ409631.1) and United Kingdom (KJ409633.1). We used species specific primers ITS5（5'-GGAAGTAAAAGTCGTAACAAGG-3'）and PITSr3 (5'AGCGCAGACATGCCGCAA-3') to amplify the product (Bulman & Marshall 1997; EPPO 2017). A single 434bp fragment was obtained from Hezhang and Weining populations. A host test for the Hezhang and Weining populations were performed by inoculating 1,000 eggs per plant of varieties Qingshu 9, Huize 2 and Hezuo 88 grown in the pots containing 800 cm3 of sterilized soil (soil: sand ratio was 3:1), and four replications were tested in greenhouse under 16 h light, 22°C in the day and 8 h dark in the night. At 90 days post inoculation, 32.6 ± 7, 31.2 ± 8, and 29.5 ± 8 females and cysts were extracted from the infected roots and soils of the varieties Qingshu 9, Huize 2 and Hezuo 88, respectively. No females and cysts were observed on the control plants. The trial indicated that potato cultivars Qingshu 9, Huize 2 and Hezuo 88 are hosts for the Hezhang and Weining populations of Globodera rostochiensis. To the best of our knowledge, this is the first detection of potato cyst nematode Globodera rostochiensis in Guizhou Province, China.

Rapid Diagnosis and Visual Detection of Potato Cyst Nematode (Globodera rostochiensis) Using Recombinase Polymerase Amplification Combination with Lateral Flow Assay Method (RPA-LFA)

Globodera rostochiensis is an important quarantine pest, it causes serious potato yield losses annually. Reliable and rapid molecular detection of G. rostochiensis is pivotal to effective early disease diagnosis and managements. Herein, recombinase polymerase amplification integrated with lateral flow assays method (RPA-LFA) was developed to target the internal transcribed spacer of nuclear ribosomal DNA (ITS rDNA) of the golden cyst nematode (G. rostochiensis), which allowed for the rapid diagnosis and detection of this nematode from crude extracts of cysts and juveniles within 30 min. Sensitivity test results showed that 10−1 single juvenile and 10−3 single cyst can be reliably detected. Moreover, the RPA-LFA method can directly diagnose and detect G. rostochiensis from infested field soil. This is the first RPA-LFA method for diagnosis G. rostochiensis, it is a fast, accurate, and sensitive detection method and can be developed for detection of G. rostochiensis in fields and laboratories lacking large instrument and equipment.

Cloning and Molecular Characterization of Hsp Genes from Anoplophora glabripennis and Their Responses to Cold Stress.

Anoplophora glabripennis (Agla) is an important global quarantine pest due to its highly destructive impacts on forests. It is widely distributed in many countries in Asia, Europe, and North America. The survival of A. glabripennis larvae has been facilitated by its high adaptability to low temperature. When insects are subjected to temperature stress, heat shock proteins (Hsps) limit cell damage and improve cell tolerance via their protein folding, localization, and degradation activities. However, the temperature adaptation mechanisms of A. glabripennis Hsps remain unclear. In this study, four A. glabripennis Hsp genes, AglaHsp20.43, AglaHsp71.18, AglaHsp82.09, and AglaHsp89.76, were cloned. Sequence analysis showed that all four Hsps had specific conserved domains. Phylogenetic analysis revealed that Hsps from different subfamilies were evolutionarily conserved, and that AglaHsps were highly similar to those of Coleoptera species. Protein expression vectors (pET30a-AglaHsps) were constructed and used to express AglaHsps in E. coli, where all four proteins were expressed in inclusion bodies. Western blot analysis showed that AglaHsps were expressed at a range of temperatures, from −10 °C to 25 °C. AglaHsp82.09 and AglaHsp89.76 showed high expressions with treatment at 0 °C. Our results will facilitate clarification of the molecular mechanisms underlying A. glabripennis responses to environmental stress.

The effectiveness of mass trapping of Ceratitis capitata (Wiedemann,1824) (Diptera: Tephritidae) in peach orchards in İzmir, Aydın and Mersin

Ceratitis capitata karantina zararlısı ve toleransı sıfır olarak kabul edilen bir türdür. Üreme kapasitesinin yüksek olması ve doğrudan meyvede zarar yapması nedeniyle zararlı ile mücadele yürütülmeden üretim yapmak mümkün olamamaktadır. Kimyasal mücadeleye alternatif olarak biyoteknik mücadele yöntemi olan kitle halinde tuzakla yakalama, zararlının çok yüksek olmayan popülasyonlarında başarılı sayılan bir mücadele şeklidir. Tuzaklar ile mücadelede farklı besin cezbedici materyaller kullanılarak bireylerin tuzağa çekilmesi sonucu öldürülmesi sağlanmaktadır. Bu çalışma, Ege ve Akdeniz Bölgelerinde şeftali bahçelerinde eş zamanlı yürütülmüş ve bölgeler arasındaki popülasyon farklılığına göre tuzakların etkinliğinin belirlenmesi amaçlanmıştır. Denemeler 2018 yılında Kuşadası (Aydın), Selçuk (İzmir) ve Erdemli (Mersin) ilçelerinde yürütülmüştür. Zararlının feromon tuzaklar ile popülasyon değişimi ve kitle halinde yakalama tuzaklarının etkinliği saptanmıştır. Mücadele tuzağı olarak Decis Trap (Bayer) isimli ürün kullanılarak tuzağın etkinlikleri saptanmıştır. Temmuz ayında Kuşadası, Selçuk ve Erdemli ilçelerinde sırasıyla 4.8, 149.1 ve 166.9 adet/tuzak ergin birey yakalamıştır. Buna göre Kuşadası ilçesinde %94.19, Selçuk ilçesinde %95.6 oranlarında etkili bir başarı sağlarken Erdemli ilçesinde %56.34 oranında etkili olmuştur. Akdeniz Bölgesi popülasyonunun Ege Bölgesine göre daha yüksek seyretmesi ve şeftali üretim sezonun daha uzun sürmesinden dolayı istenilen düzeyde başarı sağlanamadığı belirlenmiştir.

Climate change effects on the global distribution and range shifts of citrus longhorned beetle Anoplophora chinensis

AbstractThe citrus longhorned beetle (CLB) (Anoplophora chinensis) is an important quarantine pest of main Citrus crops. Its potential distribution and invasion under climate change scenarios have important economic implications for many countries. We used the ensemble maps from both the CLIMEX and MaxEnt models to obtain CLB’s potential distribution in climate‐suitable regions under historical and future climate conditions. Global broadleaved forest cover was overlaid with CLB’s ensemble distributions to further assess the effects of host range. The ensemble models’ projected climate‐suitable regions by 2040–2060 and 2060–2070 under different emission scenarios were used to analyze range shifts. Our results indicate that projected climate‐suitable regions of the CLIMEX are much wider than that of the MaxEnt, but both of them would lose areas with climate change. The global ensemble distributions of CLB concentrated in eastern Asia, central and western Europe and eastern North America, and would shift northward in the future time. Broadleaved forests would cover most of the projected climate‐suitable regions, which provide essential hosts for CLB’s establishment. The ensemble predictive results from the correlative model and mechanistic model highlight the necessity of increasing control, monitoring and quarantine efforts on the pest in the threatened areas.