Combined lethal toxicity, biochemical responses, and gene expression variations induced by tebuconazole, bifenthrin and their mixture in zebrafish (Danio rerio).

Pesticides commonly occur as mixtures in an aqueous environment, causing deleterious effects on human health and the environment. However, the mechanism underlying the combined effects on aqueous organisms remains largely unknown, especially at low concentrations. In the current study, we inspected the interactive toxicity of tebuconazole (TEB), a triazole fungicide, and bifenthrin (BIF), a pyrethroid insecticide, to zebrafish (Danio rerio) using various toxicological assays. Our data revealed that the 96 h-LC50 (lethal concentration 50) values of BIF to fish at different life periods (embryonic, larval, juvenile, and adult periods) ranged from 0.013 (0.011-0.016) to 0.41 (0.35-0.48) mg a.i. L-1, which were lower than that of TEB ranging from 1.1 (0.88-1.3) to 4.8 (4.1-5.7) mg a.i. L-1. Combination of TEB and BIF induced synergetic acute toxicity to embryonic fish. Activities of T-SOD, POD, and GST were distinctly altered in most individual and joint administrations. Expressions of 16 genes associated with oxidative stress, cellular apoptosis, immune system, and endocrine system at the mRNA level were evaluated, and the information revealed that embryonic zebrafish were impacted by both individual compounds and their combinations. Six genes (cas9, P53, gr, TRα, IL-8, and cxcl-clc) exhibited greater changes when exposed to pesticide mixtures. Therefore, the joint effects induced by the pesticides at low concentrations should be considered in the risk assessment of mixtures and regulated as priorities for mixture risk management in the aqueous ecosystem. More research is needed to identify the threshold concentrations of the realistic pesticide mixtures above which synergistic interactions occur.

Variations of enzymatic activity and gene expression in zebrafish (Danio rerio) embryos co-exposed to zearalenone and fumonisin B1.

The natural co-occurrence of multiple mycotoxins has been reported in cereals and cereal products worldwide. Even though the dietary exposure to mycotoxins constitutes a serious human health, most reports are limited to the toxic effect of individual mycotoxins. The purpose of the present study was to assess the combined toxic effects of zearalenone (ZEN) and fumonisin B1 (FB1) and the potential interaction of their mixture on zebrafish (Danio rerio) embryos. Our results showed that ZEN possessed the higher toxicity to embryonic zebrafish (7-day LC50 value of 0.78 mg a.i. L-1) compared with FB1 (7-day LC50 value of 227.7 mg a.i. L-1). The combination of ZEN and FB1 exerted an additive effect on zebrafish embryos. Meanwhile, the activities of antioxidant CAT, caspase-3, and detoxification enzyme CYP450, as well as the expressions of six genes (Mn-sod, cas9, bax, cc-chem, ERα, and crh) associated with oxidative stress, cellular apoptosis, immune system, and endocrine system were prominently altered in the mixture exposure compared with the corresponding single treatment group of ZEN or FB1. Taken together, the regulatory standards of mycotoxins in food and feed should be updated based on the mixture effects of mycotoxins, and there is an increased need on effective detoxification methods for controlling and reducing the toxicity of multiple mycotoxins in animal feed and throughout the food supply chain.

Frontalin pheromone biosynthesis in the mountain pine beetle, Dendroctonus ponderosae, and the role of isoprenyl diphosphate synthases.

The mountain pine beetle (Dendroctonus ponderosae Hopkins) is the most destructive pest of western North American pine forests. Adult males produce frontalin, an eight-carbon antiaggregation pheromone, via the mevalonate pathway, as part of several pheromones that initiate and modulate the mass attack of host trees. Frontalin acts as a pheromone, attractant, or kairomone in most Dendroctonus species, other insects, and even elephants. 6-Methylhept-6-en-2-one, a frontalin precursor, is hypothesized to originate from 10-carbon geranyl diphosphate (GPP), 15-carbon farnesyl diphosphate (FPP), or 20-carbon geranylgeranyl diphosphate (GGPP) via a dioxygenase- or cytochrome P450-mediated carbon-carbon bond cleavage. To investigate the role of isoprenyl diphosphate synthases in pheromone biosynthesis, we characterized a bifunctional GPP/FPP synthase and a GGPP synthase in the mountain pine beetle. The ratio of GPP to FPP produced by the GPP/FPP synthase was highly dependent on the ratio of the substrates isopentenyl diphosphate and dimethylallyl diphosphate used in the assay. Transcript levels in various tissues and life stages suggested that GGPP rather than GPP or FPP is used as a precursor to frontalin. Reduction of transcript levels by RNA interference of the isoprenyl diphosphate synthases identified GGPP synthase as having the largest effect on frontalin production, suggesting that frontalin is derived from a 20-carbon isoprenoid precursor rather than from the 10- or 15-carbon precursors.

Cloning and characterization of two EcR isoforms from Japanese pine sawyer, Monochamus alternates.

The ecdysone receptor (EcR) is the hormonal receptor of ecdysteroids, which regulates insect growth and development. In this study, we cloned and characterized two isoforms of EcR in Monochamus alternates named MaEcR A and MaEcR B. The cDNAs of MaEcR A and MaEcR B have open repeating frames of 1,695 and 1,392 bp, respectively. The deduced proteins have the same C-terminal sequence and varied in N-terminal, and are consistent with reports on other insect species, particularly with the receptor of another coleopteran, Tribolium castaneum. The isoform-specific developmental expression profile of EcR in the epidermis and the midgut were analyzed with quantitative real-time reverse-transcriptase polymerase chain reaction in the pupal stage. RNA interference (RNAi) with common or isoform-specific regions induced developmental stagnation. When treated in the later larval stage, RNAi with either the common sequence or an EcR A specific sequence caused more severe effects and most larvae died prior to adulthood. The EcR B specific sequence caused less severe effects and about half of the treated larvae became adults, but some showed developmental defects. RNAi with both isoforms at early pupal stage attenuated the expression of 20E-regulated genes E74, E75, and HR3. The study demonstrates the role of EcR in the transduction of ecdysteroid response in Monochamus alternatus.

Joint toxic effects of thiamethoxam and flusilazole on the adult worker honey bees (Apis mellifera L.).

Insect pollinators are routinely exposed to a complex mixture of many pesticides. However, traditional environmental risk assessment is only carried out based on ecotoxicological data of single substances. In this context, we aimed to explore the potential effects when worker honey bees (Apis mellifera L.) were simultaneously challenged by thiamethoxam (TMX) and flusilazole (FSZ). Results displayed that TMX possessed higher toxicity to A. mellifera (96-h LC50 value of 0.11 mg a. i. L-1) than FSZ (96-h LC50 value of 738 mg a. i. L-1). Furthermore, the mixture of TMX and FSZ exhibited an acute synergistic impact on the pollinators. Meanwhile, the activities of SOD, caspase 3, caspase 9, and PPO, as well as the expressions of six genes (abaecin, dorsal-2, defensin-2, vtg, caspase-1, and CYP6AS14) associated with oxidative stress, immune response, lifespan, cell apoptosis, and detoxification metabolism were noteworthily varied in the individual and mixture challenges than at the baseline level. These data revealed that it is imminently essential to investigate the combined toxicity of pesticides since the toxicity evaluation from individual compounds toward honey bees may underestimate the toxicity in realistic conditions. Overall, the present results could help understand the potential contribution of pesticide mixtures to the decline of bee populations.

Mixture toxicities of tetrachlorantraniliprole and tebuconazole to honey bees (Apis mellifera L.) and the potential mechanism.

The extensive use of pesticides has negative effects on the health of insect pollinators. Although pollinators in the field are seldom exposed to individual pesticides, few reports have assessed the toxic impacts of pesticide combinations on them. In this work, we purposed to reveal the combined impacts of tetrachlorantraniliprole (TET) and tebuconazole (TEB) on honey bees (Apis mellifera L.). Our data exhibited that TET had greater toxicity to A. mellifera (96-h LC50 value of 298.2 mg a.i. L-1) than TEB (96-h LC50 value of 1,841 mg a.i. L-1). The mixture of TET and TEB displayed acute synergistic toxicity to the pollinators. Meanwhile, the activities of CarE, CYP450, trypsin, and sucrase, as well as the expressions of five genes (ppo, abaecin, cat, CYP4G11, and CYP6AS14) associated with immune response, oxidative stress, and detoxification metabolism, were conspicuously altered when exposed to the mixture relative to the individual exposures. These results provided an overall comprehension of honey bees upon the challenge of sublethal toxicity between neonicotinoid insecticides and triazole fungicides and could be used to assess the intricate toxic mechanisms in honey bees when exposed to pesticide mixtures. Additionally, these results might guide pesticide regulation strategies to enhance the honey bee populations.

Mixture toxic effects of thiacloprid and cyproconazole on honey bees (Apis mellifera L.).

Pesticide exposure remains one of the main factors in the population decline of insect pollinators. It is urgently necessary to assess the effects of mixtures on pollinator risk assessments because they are often exposed to numerous agrochemicals. In the present study, we explored the mixture toxic effects of thiacloprid (THI) and cyproconazole (CYP) on honey bees (Apis mellifera L.). Our findings revealed that THI possessed higher acute toxicity to A. mellifera (96-h LC50 value of 216.3 mg a.i. L-1) than CYP (96-h LC50 value of 601.4 mg a.i. L-1). It's worth noting that the mixture of THI and CYP exerted an acute synergistic effect on honey bees. At the same time, the activities of detoxification enzyme cytochrome P450s (CYP450s) and neuro target enzyme Acetylcholinesterase (AChE), as well as the expressions of seven genes (CRBXase, CYP306A1, CYP6AS14, apidaecin, defensing-2, vtg, and gp-93) associated with detoxification metabolism, immune response, development, and endoplasmic reticulum stress, were significantly altered in the combined treatment compared with the corresponding individual exposures of THI or CYP. These data indicated that a mixture of THI and CYP could disturb the physiological homeostasis of honey bees. Our study provides a theoretical basis for in-depth studies on the impacts of pesticide mixtures on the health of honey bees. Our study also provides important guidance for the rational application of pesticide mixtures to protect pollinators in agricultural production effectively.

Interacted toxic mechanisms of ochratoxin A and tricyclazole on the zebrafish (Danio rerio).

Despite the current efforts to identify the mixtures of chemical pollutants, they are often "binned" into their corresponding pollutant groups. Limited studies have investigated complex mixtures of chemical pollutants co-occurring across different groups. The combined toxic impacts of several substances become a critical consideration in toxicology because chemical combinations can exert a greater deleterious effect than the single components in the mixture. In the current work, we assessed the joint impacts of ochratoxin A and tricyclazole on the zebrafish (Danio rerio) embryos and explored their underlying signaling pathways. Ochratoxin A displayed higher toxicity than tricyclazole, with a 10-day LC50 of 0.16 mg L-1, whereas that for tricyclazole was 1.94 mg L-1. The combination of ochratoxin A and tricyclazole exhibited a synergistic impact on D. rerio. The activities of detoxification enzymes GST and CYP450, as well as apoptosis-associated enzyme caspase 3, were distinctly changed in most individual and mixture exposures comparing to the untreated group. Upon both individual and mixture exposures, more dramatic variations were detected in the expressions of nine genes, such as the apoptosis genes cas3 and bax, antioxidant gene mn-sod, immunosuppression gene il-1ß, and the endocrine system genes trα, dio1, trß, ugtlab, and crh, compared with the untreated group. These findings suggested that the simultaneous exposure to low doses of mycotoxins and pesticides in food commodities was more toxic than predicted from the individual chemicals. Considering the frequent co-occurrence of mycotoxins and pesticides in the diet, this synergy should be considered in future assessments.

Germline transformation of the silkworm Bombyx mori L. by sperm-mediated gene transfer.

The domesticated silkworm Bombyx mori L. has important roles in basic biological research and applied science. To explore the practical use of transgenic technology in agricultural silkworm varieties, we fused the neomycin-resistance gene (Neo(R)) and the green fluorescent protein gene (gfp) into the piggyBac-based transposon vector and transduced it into silkworms by sperm-mediated gene transfer (SMGT). Fluorescence observation indicated the positive rate of G0 egg-batches is 72.7%. After screening against the antibiotic G418, development of individual larvae in the same brood showed significant size differences. PCR detection indicated the existence of gfp and Neo(R) and confirmed the positive rate of transgenesis as 0.47%. Southern blot analysis confirmed the presence of the exogenous genes in the genome of G7 larvae. These results show that our strategy is practical and markedly improves the efficiency of SMGT.

Joint toxic effects of phoxim and lambda-cyhalothrin on the small yellow croaker (Larimichthys polyactis).

Although pesticides commonly exist as combinations in real-life situations of the aquatic ecosystem, the impact of the toxicity of their mixtures has remained largely unclear. In this study, we investigated the combined effects of two neurotoxic pesticides, including one organophosphate insecticide phoxim (PHO) and one pyrethroid insecticide lambda-cyhalothrin (LCY), on the embryos of the small yellow croaker (Larimichthys polyactis), and their potential pathways. LCY exhibited higher toxicity relative to PHO, with a 72-h LC50 value of 0.0074 mg a.i. L-1, while the corresponding value for PHO was 0.12 mg a.i. L-1. The mixture of PHO and LCY exerted a synergistic effect on the embryos of L. polyactis. The activities of antioxidant enzyme CAT and apoptotic enzyme caspase 3 were substantially changed in most single and combined exposure groups relative to the baseline value. Under both single and combined exposures, more significant changes were found in the mRNA expression of five genes, including the immunosuppression gene ngln2, the apoptosis gene P53, the endocrine system gene cyp19a1b, as well as neurodevelopment genes of ap and acp2, relative to the baseline value. Furthermore, the non-target metabolomic analysis demonstrated that hundreds of differential metabolites, including two bile acids (taurodeoxycholic acid and tauroursodeoxycholic acid), were significantly increased in the exposure groups. The bile acids were closely associated with the gut microbiota, and 16S rRNA sequencing results demonstrated dysfunction of the gut microbiota after exposure, especially in the combined exposure group. Our findings indicated that there might be a potential risk connected to the co-occurrence of these two pesticides in aquatic vertebrates. Consequently, future ecological risk assessments should incorporate synergistic mixtures because the current risk assessments do not consider them.

Combined toxic impacts of thiamethoxam and four pesticides on the rare minnow (Gobiocypris rarus).

To examine pesticide mixture toxicity to aqueous organisms, we assessed the single and combined toxicities of thiamethoxam and other four pesticides (chlorpyrifos, beta-cypermethrin, tetraconazole, and azoxystrobin) to the rare minnow (Gobiocypris rarus). Data from 96-h semi-static toxicity assays of various developmental phases (embryonic, larval, juvenile, and adult phases) showed that beta-cypermethrin, chlorpyrifos, and azoxystrobin had the highest toxicities to G. rarus, and their LC50 values ranged from 0.0031 to 0.86 mg a.i. L-1, from 0.016 to 6.38 mg a.i. L-1, and from 0.39 to 1.08 mg a.i. L-1, respectively. Tetraconazole displayed a comparatively high toxicity, and its LC50 values ranged from 3.48 to 16.73 mg a.i. L-1. By contrast, thiamethoxam exhibited the lowest toxic effect with LC50 values ranging from 37.85 to 351.9 mg a.i. L-1. Rare minnow larvae were more sensitive than embryos to all the pesticides tested. Our data showed that a pesticide mixture of thiamethoxam-tetraconazole elicited synergetic toxicity to G. rarus. Moreover, pesticide mixtures containing beta-cypermethrin in combination with chlorpyrifos or tetraconazole also had synergetic toxicities to fish. The majority of pesticides are presumed to have additive toxicity, while our data emphasized that the concurrent existence of some chemicals in the aqueous circumstance could cause synergetic toxic effect, leading to severe loss to the aqueous environments in comparison with their single toxicities. Thence, the synergetic impacts of chemical mixtures should be considered when assessing the ecological risk of chemicals.

Mixture toxicity of thiophanate-methyl and fenvalerate to embryonic zebrafish (Danio rerio) and its underlying mechanism.

Though pesticide mixtures can reflect the real-life situation in the water ecosystem, the quantification of their toxicity is still not fully understood. Combined effects of thiophanate-methyl (THM) and fenvalerate (FEN) on embryonic zebrafish (Danio rerio) and underlying mechanism were conducted in this study. Results showed that the 96-h LC50 values of THM to D. rerio at different growth periods ranged from 12.1 to 26.1 mg L-1, which were lower in comparison with those of FVR ranging from 0.025 to 2.8 mg L-1. Mixture of THM and FVR exhibited a synergetic response to zebrafish embryos. Activities of Cu/Zn-SOD, POD, caspase 3 and caspase 9 were significantly different in most of single and mixture administrations compared with the control group. In addition, five genes (P53, Cu/Zn-sod, crh, ERα and IL-8) associated with oxidative stress, cellular apoptosis, immune system and endocrine system showed greater variations of expressions when administrated to pesticide mixtures compared with single chemicals. Our experimental results exhibited that mixtures of thiophanate-methyl and fenvalerate produced higher toxicity towards aqueous vertebrates than when determined singly. Collectively, upcoming environmental risk assessments established according to single administrations might not be enough to protect the water ecosystem.

Changes of enzyme activity and gene expression in embryonic zebrafish co-exposed to beta-cypermethrin and thiacloprid.

Pesticides often occur as mixtures of complex compounds in water environments, while most of studies only focus on the toxic effects of individual pesticides with little attention to the joint toxic effects. In the present study, we aimed to the mixture toxicity of beta-cypermethrin (BCY) and thiacloprid (THI) to zebrafish (Danio rerio) employing multiple toxicological endpoints. Results displayed that the 96-h LC50 values of BCY to D. rerio at various developmental stages ranged from 2.64 × 10 (1.97 × 10-3.37 × 10) to 6.03 × 103 (4.54 × 103-1.05 × 104) nM, which were lower than those of THI ranging from 2.97 × 104 (1.96 × 104-4.25 × 104) to 2.86 × 105 (2.19 × 105-5.87 × 105) nM. Mixtures of BCY and THI exhibited synergistic response in embryonic zebrafish. Meanwhile, the enzyme activities of antioxidants (CAT and SOD) and detoxification enzyme (CarE), endogenous T-GSH and MDA contents, as well as gene expressions (tsh, crh, cxcl and bax) involved in oxidative stress, cellular apoptosis, immune system and endocrine system were obviously changed in the mixture exposure compared with the respective BCY or THI treatment. Consequently, the increased toxicity of pesticide mixture suggested that the toxicological data acquired from individual pesticide tests might underrate the toxicity risk of pesticides that actually arise in the real environment. Taken together, our present study provided evidence that mixture exposure of BCY and THI could induce additional toxic effect compared with their respective individual pesticides on D. rerio, offering valuable insights into the toxic mechanism of pesticide mixture.

Combined toxic effects of fludioxonil and triadimefon on embryonic development of zebrafish (Danio rerio).

Pesticides scarcely exist as individual compounds in the water ecosystem, but rather as mixtures of multiple chemicals at relatively low concentrations. In this study, we aimed to explore the mixture toxic effects of fludioxonil (FLU) and triadimefon (TRI) on zebrafish (Danio rerio) by employing different toxicological endpoints. Results revealed that the 96-h LC50 values of FLU to D. rerio at multiple developmental stages ranged from 0.055 (0.039-0.086) to 0.61 (0.33-0.83) mg L-1, which were less than those of TRI ranging from 3.08 (1.84-5.96) to 9.75 (5.99-14.78) mg L-1. Mixtures of FLU and TRI exerted synergistic effects on embryonic zebrafish. Activities of total superoxide dismutase (T-SOD) and catalase (CAT) were markedly altered in most of the individual and pesticide mixture treatments compared with the control. The expressions of 16 genes involved in oxidative stress, cellular apoptosis, immune system and endocrine system displayed that embryonic zebrafish were affected by the individual pesticides and their mixtures, and greater variations of four genes (ERɑ, Tnf, IL and bax) were found when exposed to pesticide mixtures compared with their individual compounds. Therefore, more studies on mixture toxicities among different pesticides should be taken as a priority when evaluating their ecological risk.

[Cloning and analysis of adenine nucleotide translocase gene in Helicoverpa armigera].

A cDNA clone encoding the ADP/ATP translocase in Helicoverpa armigera has been identified by RT-PCR, 5'and 3'RACE methods. Sequence analysis shows that it is 1,190 bp long and contains a single open reading frame (ORF, 133-1,033 bp) encoding a protein of 300 amino acids (GenBank submission number, AY253868). The protein has a 22 aa signal peptide on its N-terminal, which leads the protein locating onto the inner membrane of the mitochondria. It also has three conserved domains of the mitochondrial carrier protein forming a channel to exchange ATP and ADP energy molecule through the inner membrane of the mitochondria. It shows extensive similarities to the known ADP/ATP translocase poly-peptides. The ADP/ATP translocase similarity was up to 90% in the Lepidoptera.

[Agrobacterium tumefaciens-mediated transformation of the pathogen of Botrytis cinerea].

Employing gfp as a reporter gene and hygromycin gene (hph) as a selection marker, the recombinant vector pKPG was constructed and transformed into fresh conidia of Botrytis cinerea via Agrobacterium tumefaciens. Transformants were identified by PCR analysis of gfp and hph cassette, green fluorescence observation with microscope and Southern hybridization. Results confirmed that target genes were successfully integrated into the genome of Botrytis cinerea.

Evaluation of joint effects of cyprodinil and kresoxim-methyl on zebrafish, Danio rerio.

Aquatic organisms are usually exposed to a mixture of pesticides instead of individual chemicals. However, risk assessment of pesticides is traditionally based on toxicity data of individual compounds. In this study, we aimed to examine the joint toxicity of two fungicides cyprodinil (CYP) and kresoxim-methyl (KRM) to zebrafish (Danio rerio) using a systematic experimental approach. Results from 96-h semi-static test indicated that the LC50 values of KRM to D. rerio at multiple life stages (embryonic, larval, juvenile and adult stages) ranged from 0.034 (0.015-0.073) to 0.61 (0.39-0.83) mg a.i. L-1, which were higher than those of CYP ranging from 1.05 (0.88-1.52) to 4.42 (3.24-6.02) mg a.i. L-1. Pesticide mixtures of CYP and KRM exhibited synergistic effect on embryonic zebrafish. The activities of carboxylesterase (CarE) and cytochrome P450 (Cyp450) were significantly altered in most of the individual and combined exposures compared with the control group. The expressions of seven genes (Mnsod, cyp17, crhr 2, crh, gnrhr 4, gnrhr 1 and hmgrb) were significantly altered upon exposure to combined pesticides compared with their individual pesticides. Collectively, these findings suggested joint effects should be considered in the risk assessment of pesticides and development of water quality criteria for the protection of aquatic environment.

[Cloning and mapping of DpKettin gene on Z chromosome of the pine caterpillar by real-time quantitative PCR].

The ratio of Z chromosome number to autosomal chromosome (A) is different between male and female lepidoptera insects. In males: 2Z : 2A=1, and in females: Z : 2A=0.5. The ratio of the copies of gene (such as K) on Z chromosome number to the copies of gene (such as N) on autosomal chromosome (K/N) is also different between male and female. In males: 2K : 2N =1, and in females: K : 2N=0.5. The DpKettin gene of the pine caterpillar Dendrolimus punctatus was cloned with the primers designed according to the sequences of the BmKettin of the silkworm and the HaKettin of the cotton bollworm. The ratio of copies between DpKettin and ANT (adenine nucleotide translocator) in male and female was determined by the real-time quantitative PCR technique. They were 1.0 in males and 0.5 in females. These ratios were equal to the ratios of the copies of gene on Z chromosome number to the copies of gene on autosomal chromosome. It indicates that DpKettin is located on Z chromosome in the genome of the pine caterpillar.

[Cloning and structural analysis of MLP in the silkworm, Bombyx mori].

The LIM domain is found in a wide variety of eukaryotic proteins that regulate gene expression and cell differentiation during development. Muscle LIM protein (MLP) gene in Bombyx mori has been cloned by blasting its EST database and PCR test in present report. The resulting sequence covers 2 327 bp of cDNA (GenBank accession No. DQ311195). It has a complete open reading fragment and encodes a 494 amino acid protein. Genomic DNA sequence contains 11 exons and 10 introns, with intron splicing following the GT-AG rule. M.W. and PI of the predicted MLP in Bombyx mori are 53.03 kDa and 8.29 respectively. A single LIM domain linked to a glyscine-rich region is found in a previously deposited LIM protein (AAR23823) in Bombyx mori. MLP identified in this report encodes a protein with five tandem LIM-glycine modules. The two LIM proteins could be produced by alternative splicing and both are probably involved in muscle cell differentiation. This work provides foundation for further research on the in vivo function of MLP.

Complete mitochondrial genome of the Japanese pine sawyer, Monochamus alternatus (Coleoptera: Cerambycidae).

We determined the complete mitochondrial genome of the Japanese pine sawyer Monochamus alternatus Hope (Coleoptera: Cerambycidae), which is a major forest pest in Asia. The genome is 15,874 bp in length containing 37 typical animal mitochondrial genes and one non-coding A+T-rich region. Its gene content and order are typical of other coleopteran mitochondrial genomes described to date. All protein-coding genes (PCGs) are initiated by ATN codons. Eight PCGs use complete stop codons TAG or TAA, whereas other PCGs end with a single T. All tRNA genes show typical secondary cloverleaf structures except for tRNA(Ser(AGN)), which lacks the dihydrouridine (DHU) arm. The large non-coding A+T-rich region of 1249 bp contains a 14 bp-long poly-T stretch and two microsatellite-like (AT)(TA)7 and (TA)8 elements.