Ectropis Obliqua Research Articles

Physicochemical Basis and Comparison of Two Type II Sex Pheromone Components Binding with Pheromone-Binding Protein 2 from Tea Geometrid, Ectropis obliqua

Lepidopteran geometrid moth can produce complex Type II sex pheromone components to attract males and trigger mating behavior. Although several sex pheromone components have been identified, it remains unclear whether their physicochemical roles in sex pheromone sensing are the same. Therefore, we utilized tea geometrid ( Ectropis obliqua) as an example model to investigate and compare the physicochemical basis of two key Type II sex pheromone components, cis-6,7-epoxy-(3Z,9Z)-3,9-octadecadiene ( Z3 Z9-6,7-epo-18:Hy) and ( Z, Z, Z)-3,6,9-octadecatriene (Z3Z6Z9-18:Hy), interacting with pheromone-binding protein 2 ( EoblPBP2) from E. obliqua. Multispectral, thermodynamic, docking, and site-directed mutagenesis indicated that the major sex pheromone component Z3Z9-6,7-epo-18:Hy is more susceptible to pH-tuned than the minor component Z3Z6Z9-18:Hy, whereas Z3Z6Z9-18:Hy seems to be more susceptible to temperature and amino acid mutations than Z3Z9-6,7-epo-18:Hy. Our study suggests that different components of Type II sex pheromone play different binding characters under specific conditions in the physicochemical behavior. This deeply supplements the theoretical knowledge of Type II pheromones involved in the recognition and discrimination in the Lepidopteran sex pheromones family.

Evaluation of E-nose data analyses for discrimination of tea plants with different damage types

This study employed electronic nose (E-nose) to detect tea plants with different types of damage (undamaged, mechanically damaged, damages caused by Ectropis obliqua and Ectropis grisescens). Gas chromatography–mass spectrometry was employed as an auxiliary technique for proving the potential of E-nose detection. A new feature extraction method was applied for obtaining comprehensive features of E-nose dataset. Feature selection method based on principal component analysis (PCA) was applied for further feature selection. Four dimensionality reduction methods [PCA, locality preserving projections (LPP), kernel principal component analysis and locally linear embedding] and three classification algorithms [multilayer perceptron neural network, extreme learning machine and support vector machine (SVM)] were employed, and the best combination of dimensionality reduction method and classification algorithm was determined. The results showed that the combination of LPP and SVM was the best, and its correct discrimination rate was as high as 100%, which proved the advantage of the new signal processing method and the feasibility of E-nose in discriminating among tea plants with different types of damage.

Decreased capture of natural enemies of pests in light traps with light‐emitting diode technology

Light traps, containing a fluorescent lamp, are commonly used to control pests in Chinese tea gardens. However, owing to them emitting light with a wide wavelength range, they can also attract and kill numerous natural enemies of such pests, which can potentially threaten the ecological balance of tea gardens. The light source in traps is a key factor determining the species of trapped insects. Based on interspecific differences in the phototactic response to diverse light of narrow wavelength among insects, a light source friendly to natural enemies of pests was designed in this study using light‐emitting diode chips. First, the wavelengths corresponding to the peaks of phototactic responses of two major tea pests, Ectropis obliqua (Lepidoptera: Geometridae) (385 nm) and Empoasca onukii (Hemiptera: Cicadellidae) (420 nm), and of 10 dominant natural enemies (380 nm) in tea gardens, were confirmed by indoor bioassays, and then 385‐nm and 420‐nm light‐emitting diode chips were selected to construct the light source for traps. Furthermore, a light trap combining a conical light source with suction fan equipment was designed to increase the quantity and lethality of catches. Finally, evaluation of the performance of the designed light trap was conducted in the field, revealing that it had the desired effect, namely, trapping more tea pests as well as fewer natural enemies compared with light traps with a fluorescent lamp.

Benzothiadiazole and B-Aminobutyricacid Induce Resistance to Ectropis Obliqua in Tea Plants (Camellia Sinensis (L.) O. Kuntz).

In order to investigate the effect of benzothiadiazole (BTH) and β-aminobutyric acid (BABA) on the resistance of tea plants (Camellia sinensis) to tea geometrid (Ectropis obliqua), three levels each of benzothiadiazole (BTH) and β-aminobutyric acid (BABA) were sprayed on 10-year-old tea plants. Generally PPO and PAL activities increased with low concentrations of BTH and BABA treatments. Quantitative RT-PCR revealed a 1.43 and 2.72-fold increase in PPO gene expression, and 3.26 and 3.99-fold increase in PAL gene expression with 75 mg/L BTH and 400 mg/L BABA respectively. Analysis of hydrolysis of synthetic substrates also revealed that chymotrypsin-like enzyme activity present in larval midgut extracts was not significantly inhibited by BTH and BABA. However, proteinase activity was found to be inversely proportional to the age of tea geometrid. Larvae pupation rate decreased by 8.10, 10.81 and 21.62% when tea geometrid were fed with leaves treated with 25, 50 and 75 mg/L BTH solutions, while 100, 200 and 400 mg/L BABA solutions decreased same by 8.10, 16.21 and 13.51% respectively. Also, larvae development period delayed to 23.33 and 26.33 days with 75 mg/L BTH and 400 mg/L BABA treatments respectively. The results in this study; therefore, suggest that benzothiadiazole (BTH) and β-aminobutyric acid (BABA) play a role in inducing resistance in tea plants to tea geometrid, with the optimal effect achieved at BTH-3 (75 mg/L) and BABA-3 (400 mg/L), respectively.

The Odorant Binding Protein 6 Expressed in Sensilla Chaetica Displays Preferential Binding Affinity to Host Plants Volatiles in Ectropis obliqua.

The monophagous tea geometrid Ectropis obliqua selectively feed on tea plants, requiring the specialized chemosensory system to forage for certain host. A deep insight into the molecular basis would accelerate the design of insect-behavior-modifying stimuli. In the present study, we focused on the odorant-binding protein 6 (EoblOBP6) with the high abundance in legs transcriptome of E. obliqua moths. qRT-PCR coupled with western blot analyses revealed the dual expression pattern of EoblOBP6 in antennae and legs. Cellular immunolocalization indicated that EoblOBP6 was predominantly labeled in the outer sensillum lymph of uniporous sensilla chaetica, which is not innervated by sensory neurons. No specific staining was observed in other sensillum types. The fluorescence competition assay showed a relatively narrow binding spectrum of recombinant EoblOBP6. EoblOBP6 could not only bind with intact tea plant volatiles benzaldehyde but also display high binding ability to nerolidol and α-farnesene which are tea plant volatiles dramatically induced by herbivore infestation. Besides, EoblOBP6 tightly bound to the aversive bitter alkaloid berberine. Taken together, EoblOBP6 displayed an unusual expression in sensilla chaetica, exhibited the potential involvement in olfaction and gustation, and may play a functional role in host location of female E. obliqua moths.

Tea saponin reduces the damage of Ectropis obliqua to tea crops, and exerts reduced effects on the spiders Ebrechtella tricuspidata and Evarcha albaria compared to chemical insecticides.

BackgroundTea is one of the most economically important crops in China. However, the tea geometrid (Ectropis obliqua), a serious leaf-feeding pest, causes significant damage to tea crops and reduces tea yield and quality. Spiders are the most dominant predatory enemies in the tea plantation ecosystem, which makes them potentially useful biological control agents of E. obliqua. These highlight the need for alternative pest control measures. Our previous studies have shown that tea saponin (TS) exerts insecticidal activity against lepidopteran pests. Here, we investigate whether TS represents a potentially new alternative insecticide with no harm to spiders.MethodsWe investigated laboratory bioactivities and the field control properties of TS solution against E. obliqua. (i) A leaf-dip bioassay was used to evaluate the toxicity of TS to 3rd-instar E. obliqua larvae and effects of TS on the activities of enzymes glutathione-S-transferase (GST), acetylcholinesterase (AChE), carboxylesterase (CES) and peroxidase (POD) of 3rd-instar E. obliqua larvae in the laboratory. (ii) Topical application was used to measure the toxicity of 30% TS (w/v) and two chemical insecticides (10% bifenthrin EC and 50% diafenthiuron SC) to two species of spider, Ebrechtella tricuspidata and Evarcha albaria. (iii) Field trials were used to investigate the controlling efficacy of 30% TS against E. obliqua larvae and to classify the effect of TS to spiders in the tea plantation.ResultsThe toxicity of TS to 3rd-instar E. obliqua larvae occurred in a dose-dependent manner and the LC50 was 164.32 mg/mL. Activities of the detoxifying-related enzymes, GST and POD, increased in 3rd-instar E. obliqua larvae, whereas AChE and CES were inhibited with time by treatment with TS. Mortalities of E. tricuspidata and E. albaria after 48 h with 30% TS treatment (16.67% and 20%, respectively) were significantly lower than those with 10% bifenthrin EC (80% and 73.33%, respectively) and 50% diafenthiuron EC (43.33% and 36.67%, respectively). The highest controlling efficacy of 30% TS was 77.02% at 5 d after treatment, which showed no difference to 10% bifenthrin EC or 50% diafenthiuron SC. 30% TS was placed in the class N (harmless or slightly harmful) of IOBC (International Organization of Biological Control) categories for natural enemies, namely spiders.ConclusionsOur results indicate that TS is a botanical insecticide that has a good controlling efficacy in E. obliqua larvae, which suggests it has promise as application in the integrated pest management (IPM) envisaged for tea crops.

Functional Characteristics, Electrophysiological and Antennal Immunolocalization of General Odorant-Binding Protein 2 in Tea Geometrid, Ectropis obliqua.

As one of the main lepidopteran pests in Chinese tea plantations, Ectropis obliqua Warren (tea geometrids) can severely decrease yields of tea products. The olfactory system of the adult tea geometrid plays a significant role in seeking behaviors, influencing their search for food, mating partners, and even spawning grounds. In this study, a general odorant-binding protein (OBP) gene, EoblGOBP2, was identified in the antennae of E. obliqua using reverse transcription quantification PCR (RT-qPCR). Results showed that EoblGOBP2 was more highly expressed in the antennae of males than in females relative to other tissues. The recombinant EoblGOBP2 protein was prepared in Escherichia coli and then purified through affinity chromatography. Ligand-binding assays showed that EoblGOBP2 had a strong binding affinity for some carbonyl-containing tea leaf volatiles (e.g., (E)-2-hexenal, methyl salicylate, and acetophenone). Electrophysiological tests confirmed that the male moths were more sensitive to these candidate tea plant volatiles than the female moths. Immunolocalization results indicated that EoblGOBP2 was regionally confined to the sensilla trichoid type-II in the male antennae. These results indicate that EoblGOBP2 may be primarily involved in the olfactory activity of male E. obliqua moths, influencing their ability to sense tea leaf volatiles. This study provides a new perspective of insect GOBPs and implies that olfactory function can be used to prevent and control the tea geometrid.

Complete mitochondrial genome of the tea looper caterpillar, Ectropis obliqua (Lepidoptera: Geometridae) with a phylogenetic analysis of Geometridae

The complete mitochondrial genome (mitogenome) of the Ectropis obliqua (Lepidoptera: Geometridae) was sequenced and compared with other species of Geometridae. The mitochondrial genome of E. obliqua is 15,475bp long and contains 37 genes including 13 protein coding genes (PCGs), 22 transfer RNA genes, 2 ribosomal RNA genes and an A+T-rich region. Gene cluster trnI-trnQ-trnM rearranged to trnM-trnI-trnQ comparing with the ancestral mitogenome of insects. All tRNA genes have typical cloverleaf secondary structure except for trnS (AGN). All the sequenced mitogenomes from Geometridae have negative AT-skews and positive GC-skews except for Biston suppressaria. The synonymous and nonsynonymous substitutions ratios (Ka/Ks) indicated variable selective pressures among 13 PCGs. The phylogenetic relationship among 11 species of Geometridae was recovered based on 13 protein coding genes and 2 rRNA genes using Bayesian inference. E. obliqua is grouped with other Ennominae species and recovered to be sister-relationship with Abraxas suspecta.

Identification and Expression Patterns of Putative Diversified Carboxylesterases in the Tea Geometrid Ectropis obliqua Prout.

Carboxylesterases (CXEs) belong to a family of metabolic enzymes. Some CXEs act as odorant-degrading enzymes (ODEs), which are reportedly highly expressed in insect olfactory organs and participate in the rapid deactivation of ester pheromone components and plant volatiles. The tea geometrid Ectropis obliqua Prout produces sex pheromones consisting of non-ester functional compounds but relies heavily on acetic ester plant volatiles to search for host plants and locate oviposition sites. However, studies characterizing putative candidate ODEs in this important tea plant pest are still relatively scarce. In the present study, we identified 35 candidate EoblCXE genes from E. obliqua chemosensory organs based on previously obtained transcriptomic data. The deduced amino acid sequences possessed the typical characteristics of the insect CXE family, including oxyanion hole residues, the Ser-Glu-His catalytic triad, and the Ser active included in the conserved pentapeptide characteristic of esterases, Gly-X-Ser-X-Gly. Phylogenetic analyses revealed that the EoblCXEs were diverse, belonging to several different insect esterase clades. Tissue- and sex-related expression patterns were studied via reverse-transcription and quantitative real-time polymerase chain reaction analyses (RT- and qRT-PCR). The results showed that 35 EoblCXE genes presented a diversified expression profile; among these, 12 EoblCXEs appeared to be antenna-biased, two EoblCXEs were non-chemosensory organ-biased, 12 EoblCXEs were ubiquitous, and nine EoblCXEs showed heterogeneous expression levels among different tissues. Intriguingly, two EoblCXE genes, EoblCXE7 and EoblCXE13, were not only strongly localized to antennal sensilla tuned to odorants, such as the sensilla trichodea (Str I and II) and sensilla basiconica (Sba), but were also expressed in the putative gustatory sensilla styloconica (Sst), indicating that these two CXEs might play multiple physiological roles in the E. obliqua chemosensory processing system. This study provides the first elucidation of CXEs in the chemosensory system of a geometrid moth species and will enable a more comprehensive understanding of the functions of insect CXEs across lepidopteran species.

Evidence of Premating Isolation Between Two Sibling Moths: Ectropis grisescens and Ectropis obliqua (Lepidoptera: Geometridae)

The sex pheromones of Ectropis grisescens Warren and Ectropis obliqua Prout were both reported to contain (Z,Z,Z)-3,6,9-octadecatriene (Z3,Z6,Z9-18:H) and (Z,Z)-3,9-cis-6,7-epoxy-octadecadiene (Z3,epo6,Z9-18:H). To clarify how these two sibling geometrids maintain premating isolation, the female sex pheromones of the two species were reexamined. Gas chromatography-electroantennographic detection (GC-EAD) and gas chromatography-mass spectrometry revealed two GC-EAD-active compounds, Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H, in E. grisescens female pheromone glands as well as an additional GC-EAD-active compound, (Z,Z)-3,9-cis-6,7-epoxy-nonadecadiene (Z3,epo6,Z9-19:H), in E. obliqua female pheromone glands. Synthesized Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H elicited dose-dependent electroantennogram (EAG) responses from male antennae of both E. grisescens and E. obliqua. However, Z3,epo6,Z9-19:H only elicited dose-dependent EAG responses from E. obliqua and limited EAG responses from E. grisescens at all doses. In wind-tunnel studies, lures that contained Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H attracted E. grisescens males and had no effect on E. obliqua males. The addition of Z3,epo6,Z9-19:H to the blend of Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H strongly attracted E. obliqua males but had a limited attraction for E. grisescens males. Thus, Z3,Z6,Z9-18:H and Z3,epo6,Z9-18:H were sex pheromone components of E. grisescens, whereas Z3,Z6,Z9-18:H, Z3,epo6,Z9-18:H and Z3,epo6,Z9-19:H were sex pheromone components of E. obliqua. The presence or absence of Z3,epo6,Z9-19:H played a central role in the premating isolation of these two sibling species.

The complete mitochondrial genome of a tea geometrid, Ectropis obliqua (Lepidoptera: Geometridae)

The tea geometrid, Ectropis obliqua Prout (Lepidoptera: Geometridae), is a major pest of tea plantation and poses a considerable economic threat to tea industry. We have sequenced the complete mitochondrial genome of E. obliqua. The entire genome is 16,535 bp in length with an A + T content of 81.32% (GenBank accession No. KX827002). The tea geometrid mt genome encodes all 37 genes that are typically found in animal mt genomes, consists of 13 protein-coding genes (PCG), two ribosomal RNA genes, and 22 transfer RNA genes. The gene order is consistent with other sequenced mt genome of moths and butterflies in Ditrysia. The A + T-rich region is 1523 bp long and consisting of the motif ‘ATAGA’, a 19 bp poly-T stretch, and a tandem repeat sequence with seven 194 bp repeat units. Phylogenetic analysis was performed using 13 PCG with 16 moths showed that E. obliqua clusters with other Geometridae species.

Main predators of insect pests: screening and evaluation through comprehensive indices.

Predatory natural enemies play key functional roles in integrated pest management. However, the screening and evaluation of the main predators of insect pests has seldom been reported in the field. Here, we employed comprehensive indices for evaluating the predation of a common pest (Ectropis obliqua) by nine common spider species in Chinese tea plantations. We established the relative dominance of the spider species and their phenological overlap with the pest species, and analyzed DNA from the nine spider species using targeted real-time quantitative polymerase chain reaction to identify the residual DNA of E. obliqua. The predation rates and predation numbers per predator were estimated by the positive rates of target fragments and the residual minimum number of E. obliqua in predators' guts, respectively. The results showed that only four spider species preyed on E. obliqua, and the order of potential of the spiders to control E. obliqua from greatest to smallest was Neoscona mellotteei, Xysticus ephippiatus, Evarcha albaria and Coleosoma octomaculatum by the Z-score method. The orb-weaving spider N. mellotteei has the maximum potential as a biological control agent of E. obliqua in an integrated pest management strategy. An approach of screening and evaluating main predators of insect pests through comprehensive indices was preliminarily established. © 2017 Society of Chemical Industry.

Highly Site-Selective Epoxidation of Polyene Catalyzed by Metal–Organic Frameworks Assisted by Polyoxometalate

Ectropis obliqua Prout is one of the most severe defoliator insects of tea plants. Sex pheromones method has been widely used for monitoring and controlling the insect pest of E. obliqua Prout due to their relative safety, low cost, and environmental friendliness. However, conventional preparing sex pheromones of E. obliqua Prout has been criticized by the costly and/or low efficiency procedure. Herein, we report a series of Fe-based metal–organic frameworks (MOFs) as efficient heterogeneous catalysts for the site-selective epoxidation of 3Z,6Z,9Z-octadecatriene to its monoepoxides. Particularly, MIL-100(Fe) combined with polyoxometalate (POM) as cocatalyst appears as the best catalyst for the oxidation of 3Z,6Z,9Z-octadecatriene to a high site selectivity to the corresponding 6,7-epoxide with good yields. 3Z,6Z,9Z-octadecatriene and its corresponding 6,7-epoxide are the primary sex pheromone components of the E. obliqua Prout. Significantly, the field bioassay experiments showed that the mixed products were highly effective in attracting E. obliqua Prout male moths. This work provides a general method to fast and clean synthesis of sex pheromones and opens new avenues for the application of Fe-based MOFs in the pest control with environmental friendliness.

Physicochemical Evidence on Sublethal Neonicotinoid Imidacloprid Interacting with an Odorant-Binding Protein from the Tea Geometrid Moth, Ectropis obliqua

Nowadays the excessive usage of neonicotinoid insecticides always results in residues in Chinese tea fields. It is not clear whether the insecticide residue at the sublethal level influences the physiological processes of tea pests. Here, we provide evidence of interaction between the neonicotinoid imidacloprid and a general odorant-binding protein, EoblGOBP2, from the tea geometrid moth, Ectropis obliqua. The interacting process was demonstrated through multiple fluorescence spectra, UV absorption spectra, circular dichroism (CD) spectra, molecular docking, etc. The binding mode was determined to be static (from 300 to 310 K) and dynamic quenching (from 290 to 300 K). The binding distance was calculated to be 6.9 nm on the basis of FRET theory. According to the thermodynamic analysis, the process was mainly driven by enthalpy (ΔH < 0), and hydrogen bond and van der Waals interactions were the main driving forces in the static and dynamic binding cases, respectively. Moreover, synchronous fluorescence spectra and CD spectra analysis showed stretching of the EoblGOBP2 peptide chains with a decreasing α-helix when imidacloprid was added. Molecular docking was applied and predicted that two hydrogen bonds were formed between imidacloprid and Arg110 in the mature peptide of EoblGOBP2. Moreover, when the absolute amounts of EoblGOBP2 in the moth antennae were measured and calculated by using real-time PCR, it was estimated that imidacloprid at sublethal level (about 0.233 and 0.175 ng/male and female moth antennae, respectively) inhibited the binding of a tea volatile, E-2-hexenal, to EoblGOBP2 at about half. This study indicates that neonicotinoid insecticide at sublethal level may still affect the olfactory cognition of the tea geometrid moth to volatile compounds from tea leaves.

Characterization of candidate odorant‐binding proteins and chemosensory proteins in the tea geometrid Ectropis obliqua Prout (Lepidoptera: Geometridae)

Insects rely heavily on their sophisticated chemosensory systems to locate host plants and find conspecific mates. Although the molecular mechanisms of odorant recognition in many Lepidoptera species have been well explored, limited information has been reported on the geometrid moth Ectropis obliqua Prout, an economically important pest of tea plants. In the current study, we first attempted to identify and characterize the putative olfactory carrier proteins, including odorant-binding proteins (OBPs) and chemosensory proteins (CSPs). By analyzing previously obtained transcriptomic data of third-instar larvae, five OBPs and 14 CSPs in E. obliqua were identified. Sequence alignment, conserved motif identification, and phylogenetic analysis suggested that candidate proteins have typical characteristics of the insect OBP or CSP family. The expression patterns regarding life stages and different tissues were determined by quantitative real-time PCR. The results revealed that four transcripts (OBP2, OBP4 and CSP8, CSP10) had larvae preferential expression profiles and nine candidate genes (PBP1, OBP1 and CSP2, CSP4, CSP5, CSP6, CSP7, CSP11, and CSP13) were adult-biased expressed. Further specific tissue expression profile evaluation showed that OBP1, OBP2, OBP4, and PBP1 were highly expressed at olfactory organs, implying their potential involvement in chemical cue detection, whereas CSPs were ubiquitously detected among all of the tested tissues and could be associated with multiple physiological functions. This study provided a foundation for understanding the physiological functions of OBPs and CSPs in E. obliqua and will help pave the way for the development of a new environmental friendly pest management strategy against the tea geometrid moth.

A putative 12-oxophytodienoate reductase gene CsOPR3 from Camellia sinensis, is involved in wound and herbivore infestation responses

12-Oxophytodienoate reductase (OPR) is a key enzyme in the biosynthesis of jasmonic acid (JA), which plays an important role in plant defense responses. Although multiple isoforms of OPRs have been identified in various annual herbaceous plants, genes encoding these enzymes in perennial woody plants have yet to be fully investigated. In the tea plant, Camellia sinensis (L.), no OPR genes have been isolated, and their possible roles in tea plant development and defense mechanism remain unknown. In this study, a putative OPR gene, designated as CsOPR3, was isolated from tea plants for the first time through the rapid amplification of cDNA ends. The open reading frame of CsOPR3 is 1197bp in length, and encodes a protein of 398 amino acids. Real-time qPCR analysis revealed that CsOPR3 was expressed in different organs. In particular, CsOPR3 was highly expressed in flowers, leaves and stems but was weakly expressed in roots and seeds. CsOPR3 expression could be rapidly induced by mechanical wounding, and increased JA levels were correlated with the wound-induced CsOPR3 expression. The infestation of the tea geometrid (TG) Ectropis obliqua Prout, regurgitant derived from TG and exogenous JA application could enhance the CsOPR3 expression. Our study is the first to report that CsOPR3 plays an important role in JA biosynthesis and tea plant defense against herbivorous insects.

Delayed Mating Impacts on the Reproductive Performance ofEctropis obliqua(Lepidoptera: Geometridae)

Abstract The impact of age at which mating occurs on the mating success, fecundity, fertility, and adult longevity of Ectropis obliqua Prout (Lepidoptera: Geometridae) was determined in laboratory testing. When newly emerged males were mated with females that emerged 1, 2, 3, 4, 5, or 6 d earlier, mating success, fecundity, and fertility generally decreased with increased age of the females. Similar responses were observed when newly emerged females were mated with males that had emerged 1, 2, 3, 4, 5, 6, or 7 d earlier and when females and males of the same age were allowed to mate. Adult longevity was inversely related to age at which mating occurred.

Characterization of eleven microsatellite markers for the tea geometrid Ectropis obliqua (Lepidoptera: Geometridae)

The tea geometrid, Ectropis obliqua Prout (Lepidoptera: Geometridae), is one of the most threatening pests of tea (Camellia sinensis (L.) O. Kuntze) plants. In this study, we developed eleven new polymorphic microsatellite markers from tea geometrid by using the microsatellite motif enriched library method. Polymorphism of each locus was detected in 69 individuals from three natural populations. The number of alleles varied from 2 to 8, and the expected and observed heterozygosities ranged from 0.042 to 0.801 and from 0.042 to 0.875, respectively. Six loci had significant deviations from the Hardy–Weinberg equilibrium (HWE). These microsatellite markers will be useful for the future study of this agricultural pest in genetic diversity, population structure and evolution of resistance. This information will be allowed to establish new pest management strategies and improve the effectiveness of pest control program of this species.

Asymmetrical reproductive interference between two sibling species of tea looper: Ectropis grisescens and Ectropis obliqua.

Ectropis grisescens Warren and Ectropis obliqua (Prout) are two morphologically similar sibling species with overlapping ranges. In this study, manipulative laboratory experiments were conducted to examine the possibility of reproductive interference in sympatric populations of E. grisescens and E. obliqua and the potential consequences of the mating interaction. Our results showed that the presence of males or females of different species could incur mating interference and significant reduction of F 1 offspring. The reduction was not significant relevant to the initial relative abundance of E. grisescens and E. obliqua. Detailed observations of mating opportunity showed that female mating frequencies of both species were not significantly affected by the absolute species density, but the mating success of E. obliqua females with conspecific males depended on species ratio. In addition, adding males to the other species resulted in lower number of offspring suggesting that the males' behaviour might be linked with mating interference. Males of both E. grisescens and E. obliqua could interfere the intraspecific mating of the other species, but the impact of the mating interference differed. These combined data indicated that asymmetric reproductive interference existed in E. grisescens and E. obliqua under laboratory conditions, and the offspring of the mixed species were significantly reduced. The long term outcome of this effect is yet to be determined since additional reproductive factors such as oviposition rate and progeny survival to adulthood may reduce the probability of demographic displacement of one species by the other in overlapping niches.

Development of synthetic volatile attractant for male Ectropis obliqua moths

The tea geometrid Ectropis obliqua is one of the most serious leaf-feeding insect pests in tea (Camellia sinensis) in East Asia. Although several volatile chemicals emitted from tea plants have been reported to be attractive to E. obliqua moths, no synthetic attractants for E. obliqua moths have been developed. By measuring the behavioral responses of the moth to a series of chemicals in the lab, we found that a blend containing a ternary mixture containing (Z)-3-hexenal, (Z)-3-hexenyl hexanoate and benzyl alcohol clearly attracted to E. obliqua moths of both sex and that (Z)-3-hexenyl acetate could enhance the attractiveness of the ternary blend. Moreover, we found that the volatiles emitted from the plant-E. obliqua larva complex have the same attractiveness as: 1) the blend of volatiles containing the ternary mixture and 2) the blend containing (Z)-3-hexenyl acetate plus the ternary mixture to both male and female moths. In a field bioassay, more male moths were observed on traps that were baited with the blend containing (Z)-3-hexenyl acetate plus the ternary mixture than on control traps. Our study raises the tantalizing possibility that synthetic blends could be deployed as attractants for pests in the field.