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Glutathione S-transferases (GSTs) constitute an important multifunctional enzyme family that plays vital roles in cellular detoxification and protecting organisms against oxidative stress caused by reactive oxygen species (ROS). In this study, we isolated a GST-like gene from Apis cerana cerana (AccGSTL) and investigated its antioxidant functions under stress conditions. We found that AccGSTL belongs to the Sigma class of GSTs. Real-time quantitative PCR and western blotting analyses showed that the mRNA and protein levels of AccGSTL were altered in response to oxidative stress caused by various external stimuli. In addition, a heterologous expression analysis showed that AccGSTL overexpression in Escherichia coli (E. coli) cells enhanced resistance to oxidative stress. After AccGSTL silencing with RNA interference (RNAi) technology, the expression of some antioxidant genes was inhibited, and the enzymatic activities of POD, CAT, and SOD were decreased. In conclusion, these data suggest that AccGSTL may be involved in antioxidant defense under adverse conditions in A. cerana cerana. [ABSTRACT FROM AUTHOR]

MAP kinase phosphatase 3 (MKP3), a member of the dual-specificity protein phosphatase (DUSP) superfamily, has been widely studied for its role in development, cancer, and environmental stress in many organisms. However, the functions of MKP3 in various insects have not been well studied, including honeybees. In this study, we isolated an MKP3 gene from Apis cerana cerana and explored the role of this gene in the resistance to oxidation. We found that AccMKP3 is highly conserved in different species and shares the closest evolutionary relationship with AmMKP3. We determined the expression patterns of AccMKP3 under various stresses. qRT-PCR results showed that AccMKP3 was highly expressed during the pupal stages and in adult muscles. We further found that AccMKP3 was induced in all the stress treatments. Moreover, we discovered that the enzymatic activities of peroxidase, superoxide dismutase, and catalase increased and that the expression levels of several antioxidant genes were affected after AccMKP3 was knocked down. Collectively, these results suggest that AccMKP3 may be associated with antioxidant processes involved in response to various environmental stresses. [ABSTRACT FROM AUTHOR]

Inositol phosphate synthase (IPS) is a rate-limiting enzyme in myo-inositol biosynthesis, which can regulate stress responses in plants and animals. However, there are few studies on the function of IPS in insects, especially in Apis cerana cerana. In this study, the inositol-3-phosphate synthase 1-B gene (AccIPS1-B) was isolated from Apis cerana cerana, and its connection to antioxidant defence was investigated. The open reading frame of AccIPS1-B was 1542 bp, encoding a 513 amino acid polypeptide. Quantitative real-time PCR analysis revealed that the expression level of AccIPS1-B was highest in pupae of Apis cerana cerana, and it was expressed at higher levels in the thorax than in other tissues tested. Moreover, the expression of AccIPS1-B was significantly upregulated by abiotic stresses. The recombinant AccIPS1-B also displayed significant tolerance to cumene hydroperoxide and HgCl2. In addition, knockdown of AccIPS1-B significantly suppressed the expression of most of the antioxidant genes and decreased the antioxidant enzymatic activities of SOD, POD, and GST. Taken together, these findings indicate that AccIPS1-B may be involved in the response to antioxidant defence and development in Apis cerana cerana. [ABSTRACT FROM AUTHOR]

Clip-domain serine proteases (Clip-SPs) mediate innate immunity and embryonic development in insects. However, the function of Clip-SPs in Apis cerana cerana is little known. Here, a Clip-SP gene, AccSp1, was identified. AccSp1 was mainly detected in third and sixth day instar larvae, dark-eyed pupae, and adults (1and 30 days post-emergence). In addition, AccSp1 was expressed at its highest level in the venom gland and epidermis than tentacle, abdomen, muscle, honey sac, head, leg, chest, hemolymph, rectum, and midgut. AccSp1 was induced by 4, 24, and 44 °C; H2O2; CdCl2; HgCl2; and pesticides (paraquat, pyridaben, and methomyl) and was inhibited by UV light and cyhalothrin treatments. When adults that had been pretreated with dsRNA 6 h prior (knocking AccSp1 down) were challenged with Bacillus bombysepticus for 18 h, the survival rate of bees greatly decreased, the activity of PO (phenoloxidase) was reduced, revealing that AccSp1 may play a critical role in assisting bees to survive the microbial infection and participate in regulating PO activity. The antioxidant enzymatic activities of catalase, peroxidase, and superoxide dismutase; the contents of hydrogen peroxide and malondialdehyde; and the ratio of NADP+/NADPH were all lower in samples containing dsRNA-AccSp1 interference than in control groups, but the content of carbonyl was not significantly different. These findings suggest the knockdown of AccSp1 may influence melanization so that the antioxidant enzyme activities and the harmful metabolites decreased. These results collectively suggest that AccSp1 plays critical roles in abiotic stresses responses and resistance to pathogens. [ABSTRACT FROM AUTHOR]

Various environmental stresses, such as heat shock, heavy metals, ultraviolet (UV) radiation and different pesticides, induce a cellular oxidative stress response. The cellular oxidative stress response is usually regulated by heat shock proteins (Hsps) acting as molecular chaperones. Stress-induced phosphoprotein 1 (STIP1), one of the most widely studied co-chaperones, functions as an adaptor that directs Hsp90 to Hsp70-client protein complexes. However, the biological functions of STIP1 remain poorly understood in honeybee (Apis cerana cerana). In this study, AccSTIP1 was identified in Apis cerana cerana. AccSTIP1 transcription was found to be induced by heat (42 °C), HgCl2, H2O2 and different pesticides (emamectin benzoate, thiamethoxam, hexythiazox and paraquat) and inhibited by CdCl2, UV and kresoxim-methyl. Moreover, western blot analysis indicated that the expression profiles of AccSTIP1 were consistent with its transcriptional expression levels. The disc diffusion assay showed that chemically competent transetta (DE3) bacteria expressing a recombinant AccSTIP1 protein displayed the smaller death zones than did control bacteria after exposure to paraquat and HgCl2. The DNA nicking assay suggested that recombinant purified AccSTIP1 protected supercoiled pUC19 plasmid DNA from damage caused by a thiol-dependent mixed-function oxidation (MFO) system. After knocking down AccSTIP1 gene expression via RNA interference (RNAi), the transcript levels of antioxidation-related genes were obviously lower in dsAccSTIP1 honeybees compared with those in the uninjected honeybees. Collectively, these results demonstrated that AccSTIP1 plays an important role in counteracting oxidative stress. This study lays a foundation for revealing the mechanism of AccSTIP1 in the Apis cerana cerana antioxidant system. [ABSTRACT FROM AUTHOR]

Cyclin-dependent kinase 5 (CDK5) is an unusual CDK whose function has been implicated in protecting the central nervous system (CNS) from oxidative damage. However, there have been few studies of CDK5 in insects. In this study, we identified the AccCDK5 gene from Apis cerana cerana and investigated its role in oxidation resistance. We found that AccCDK5 is highly conserved across species and contains conserved features of the CDK5 family. The results of qPCR analysis indicated that AccCDK5 is highly expressed during the larval and pupal stages and in the adult head and muscle. We further observed that AccCDK5 is induced by several environmental oxidative stresses. Moreover, the overexpression of the AccCDK5 protein in E. coli enhances the resistance of the bacteria to oxidative stress. The activation of CDK5 requires binding to its activator. Therefore, we also identified and cloned cyclin-dependent kinase 5 regulatory subunit 1, which we named AccCDK5r1, from Apis cerana cerana. AccCDK5r1 contains a conserved cell localization targeting domain as well as binding and activation sites for CDK5. Yeast two-hybrid analysis demonstrated the interaction between AccCDK5 and AccCDK5r1. The expression patterns of the two genes were similar after stress treatment. Collectively, these results suggest that AccCDK5 plays a pivotal role in the response to oxidative stresses and that AccCDK5r1 is a potential activator of AccCDK5. [ABSTRACT FROM AUTHOR]

Serine proteinases play important roles in innate immunity and insect development. We isolated a serine proteinase gene, designated AccSp10, from the Chinese honeybees ( Apis cerana cerana). RT-qPCR and a Western blot analysis at different pupal development stages indicated that AccSp10 might be involved in melanin formation in pupae and promote pupal development. In adult workers, the expression of AccSp10 was upregulated by treatments mimicking harmful environments such as the presence of Bacillus bombysepticus, different temperatures (4, 24 and 42 °C), HgCl, HO and paraquat; the exception was treatment with VC (vitamin C), which did not upregulate AccSp10 expression. Western blot confirmed the results. A disc diffusion assay indicated that recombinant AccSp10 accelerated E. coli cell death during stimulation with harmful substances (HgCl, paraquat and cumene hydroperoxide). These findings suggest that AccSp10 may be involved in the pupal development of Chinese honeybees and protection against microorganisms and abiotic harms. [ABSTRACT FROM AUTHOR]

Vitellogenin (Vg) is a yolk precursor protein in most oviparous females. However, Vg has not been studied in the Apis cerana cerana. In this work, the Vg gene of the A. cerana cerana has been cloned and sequenced. The gene codes for a protein consisting of 1770 amino acids in seven exons with a predicted molecular mass and isoelectric point of 200 kDa and 6.46, respectively. Additionally, an 807-bp 5′-flanking region was isolated, and potential transcription factor binding sites associated with development and stress response were identified. Quantitative real-time PCR analysis showed that AccVg is highly expressed in pupae during different developmental stages. In addition, the expression of AccVg could be induced by cold (4 and 16 °C), CdCl, and pesticide treatment. Real-time quantitative PCR (RT-PCR) and western blot analyses indicated that AccVg transcription was induced by various abiotic stresses. Western blot was used to measure the expression levels of AccVg protein. Taken together, these results suggest that AccVg most likely plays essential roles in antioxidant defence and that it may be of critical importance to honey bee survival. [ABSTRACT FROM AUTHOR]

Apis cerana cerana triosephosphate isomerase ( AccTPI), a key regulatory enzyme in glycolysis and gluconeogenesis pathway, catalyzes the interconversion of glyceraldehyde 3-phosphate to dihydroxyacetone phosphate. However, its role in A. cerana cerana has not been completely clarified. In this study, a TPI gene was cloned from A. cerana cerana and named AccTPI. The open reading frame (ORF) of AccTPI is 744 bp, which encodes 247 amino acids with a predicted molecular weight and an isoelectric point of 26.8 kDa and 8.42, respectively. The expression of AccTPI was up-regulated by some abiotic stresses, while it was down-regulated by HgCl, ultraviolet light (UV), and vitamin C (VC) treatment. Finally, a disc diffusion assay revealed that recombinant AccTPI proteins can protect cells from oxidative stresses. Taken together, all of the results indicate that AccTPI may play an important role in the metabolism, in antioxidant defense, and in many biological functions in the growth and development of A. cerana cerana. [ABSTRACT FROM AUTHOR]

RNA-binding motif proteins (RBMs) belong to RNA-binding proteins that display extraordinary posttranscriptional gene regulation roles in various cellular processes, including development, growth, and stress responses. Nevertheless, only a few examples of the roles of RBMs are known in insects, particularly in Apis cerana cerana. In the present study, we characterized the novel RNA-binding motif protein 11 from Apis cerana cerana, which was named AccRBM11 and whose promoter sequence included abundant potential transcription factor binding sites that are connected to responses to adverse stress and early development. Quantitative PCR results suggested that AccRBM11 was expressed at highest levels in 1-day postemergence worker bees. AccRBM11 mRNA and protein levels were higher in the poison gland and the epidermis than in other tissues. Moreover, levels of AccRBM11 transcription were upregulated upon all the simulation of abiotic stresses. Furthermore, Western blot analysis indicated that AccRBM11 protein expression levels could be induced under some abiotic stressors, a result that did not completely in agree with the qRT-PCR results. It is also noteworthy that the expression of some genes that connected with development or stress responses were remarkably suppressed when AccRBM11 was silenced, which suggested that AccRBM11 might play a similar role in development or stress reactions with the above genes. Taken together, the data presented here provide evidence that AccRBM11 is potentially involved in the regulation of development and some abiotic stress responses. We expect that this study will promote future research on the function of RNA-binding proteins. [ABSTRACT FROM AUTHOR]

Glutathione S-transferases (GSTs) are an important family of multifunctional enzymes in aerobic organisms. They play a crucial role in the detoxification of exogenous compounds, especially insecticides, and protection against oxidative stress. Most previous studies of GSTs in insects have largely focused on their role in insecticide resistance. Here, we isolated a theta class GST gene designated AccGSTT1 from Apis cerana cerana and aimed to explore its antioxidant and antibacterial attributes. Analyses of homology and phylogenetic relationships suggested that the predicted amino acid sequence of AccGSTT1 shares a high level of identity with the other hymenopteran GSTs and that it was conserved during evolution. Quantitative real-time PCR showed that AccGSTT1 is most highly expressed in adult stages and that the expression profile of this gene is significantly altered in response to various abiotic stresses. These results were confirmed using western blot analysis. Additionally, a disc diffusion assay showed that a recombinant AccGSTT1 protein may be roughly capable of inhibiting bacterial growth and that it reduces the resistance of Escherichia coli cells to multiple adverse stresses. Taken together, these data indicate that AccGSTT1 may play an important role in antioxidant processes under adverse stress conditions. [ABSTRACT FROM AUTHOR]

Thioredoxin peroxidase (Tpx), also named peroxiredoxin (Prx), is an important peroxidase that can protect organisms against stressful environments. AccTpx4, a 1-Cys thioredoxin peroxidase gene from the Chinese honey bee Apis cerana cerana, was cloned and characterized. The AccTpx4 gene encodes a protein that is predicted to contain the conserved PVCTTE motif from 1-Cys peroxiredoxin. Quantitative real-time PCR (Q-PCR) and Western blotting revealed that AccTpx4 was induced by various oxidative stresses, such as cold, heat, insecticides, HO, and HgCl. The in vivo peroxidase activity assay showed that recombinant AccTpx4 protein could efficiently degrade HO in the presence of DL-dithiothreitol (DTT). In addition, disc fusion assays revealed that AccTpx4 could function to protect cells against oxidative stresses. These results indicate that AccTpx4 plays an important role in oxidative stress responses and may contribute to the conservation of honeybees. [ABSTRACT FROM AUTHOR]

Arginine kinases (AK) in invertebrates play the same role as creatine kinases in vertebrates. Both proteins are important for energy metabolism, and previous studies on AK focused on this attribute. In this study, the arginine kinase gene was isolated from Apis cerana cerana and was named AccAK. A 5'-flanking region was also cloned and shown to contain abundant putative binding sites for transcription factors related to development and response to adverse stress. We imitated several abiotic and biotic stresses suffered by A. cerana cerana during their life, including heavy metals, pesticides, herbicides, heat, cold, oxidants, antioxidants, ecdysone, and Ascosphaera apis and then studied the expression patterns of AccAK after these treatments. AccAK was upregulated under all conditions, and, in some conditions, this response was very pronounced. Western blot and AccAK enzyme activity assays confirmed the results. In addition, a disc diffusion assay showed that overexpression of AccAK reduced the resistance of Escherichia coli cells to multiple adverse stresses. Taken together, our results indicated that AccAK may be involved of great significance in response to adverse abiotic and biotic stresses. [ABSTRACT FROM AUTHOR]

Apoptosis-inducing factors (AIFs) play vital roles in bioenergetic and redox metabolism. Compared with studies focused on the apoptogenic function of AIF, few studies have reported on its oxidoreductase function, especially in insects. In this study, we identified a novel AIF gene AccAIF3 that was isolated from Apis cerana cerana, and investigated its expression and structural features. Quantitative real-time PCR (qRT-PCR) revealed the highest mRNA level of AccAIF3 in adult bees at 15 days. qRT-PCR results revealed that AccAIF3 transcripts were induced by cold, CdCl, HgCl, UV, pyriproxyfen and cyhalothrin, but were downregulated by ecdysone. We also compared the expression of AIF3 from Apis cerana cerana and Apis mellifera ligustica Spinola to Ascosphaera apis, and found that AIF3s may respond to Ascosphaera apis in a species-specific manner. These data suggested that AccAIF3 may play vital roles in the response to abiotic and biotic stresses and contribute to the adaptability of honeybees to adversities. [ABSTRACT FROM AUTHOR]

The adenine nucleotide translocator (ANT), which has not been well studied in hymenopteran insects, catalyzes the transport of ATP and ADP across the impermeable mitochondrial membrane and plays a key role in the regulation of reactive oxygen species (ROS) generation and cell death. In this study, we isolated and characterized a single-copy ANT gene, referred as AccANT, from Apis cerana cerana (Fabricius) (Hymenoptera: Apidae). An alignment analysis showed that the deduced AccANT amino acid sequence has high identity with other known ANTs. Based on the genomic DNA sequence, a partial 5′-flanking region was obtained and some putative cis-acting elements related to the development and stresses were also predicted. Real-time quantitative PCR analysis indicated that the accumulation of AccANT transcripts could be detected in all development stages but reached its highest level in 1-day-old adult worker honey bees. Simultaneously, AccANT was also primarily localized in the thorax tissue of adults. Moreover, AccANT expression was found to be induced in response to temperature stress and HO injection. These results suggest that AccANT may be involved in the regulation of development and in reducing ROS generation when suffering from environmental stress. [ABSTRACT FROM AUTHOR]

Glutathione S-transferases (GSTs) are members of a multifunctional antioxidant enzyme superfamily that play pivotal roles in both detoxification and protection against oxidative damage caused by reactive oxygen species. In this study, a complementary DNA (cDNA) encoding a sigma class GST was identified in the Chinese honey bee, Apis cerana cerana ( AccGSTS1). AccGSTS1 was constitutively expressed in all tissues of adult worker bees, including the brain, fat body, epidermis, muscle, and midgut, with particularly robust transcription in the fat body. Relative messenger RNA expression levels of AccGSTS1 at different developmental stages varied, with the highest levels of expression observed in adults. The potential function of AccGSTS1 in cellular defenses against abiotic stresses (cold, heat, UV, HO, HgCl, and insecticides) was investigated. AccGSTS1 was significantly upregulated in response to all of the treatment conditions examined, although the induction levels were varied. Recombinant AccGSTS1 protein showed characteristic glutathione-conjugating catalytic activity toward 1-chloro-2,4-dinitrobenzene. Functional assays revealed that AccGSTS1 could remove HO, thereby protecting DNA from oxidative damage. Escherichia coli overexpressing AccGSTS1 showed long-term resistance under conditions of oxidative stress. Together, these results suggest that AccGSTS1 is a crucial antioxidant enzyme involved in cellular antioxidant defenses and honey bee survival. [ABSTRACT FROM AUTHOR]

Oxidative stress may be the most significant threat to the survival of living organisms. Glutathione S-transferases (GSTs) serve as the primary defences against xenobiotic and peroxidative-induced oxidative damage. In contrast to other well-defined GST classes, the Omega-class members are poorly understood, particularly in insects. Here, we isolated and characterised the GSTO2 gene from Apis cerana cerana ( AccGSTO2). The predicted transcription factor binding sites in the AccGSTO2 promoter suggested possible functions in early development and antioxidant defence. Real-time quantitative PCR (qPCR) and western blot analyses indicated that AccGSTO2 was highly expressed in larvae and was predominantly localised to the brain tissue in adults. Moreover, AccGSTO2 transcription was induced by various abiotic stresses. The purified recombinant AccGSTO2 exhibited glutathione-dependent dehydroascorbate reductase and peroxidase activities. Furthermore, it could prevent DNA damage. In addition, Escherichia coli overexpressing AccGSTO2 displayed resistance to long-term oxidative stress exposure in disc diffusion assays. Taken together, these results suggest that AccGSTO2 plays a protective role in counteracting oxidative stress. [ABSTRACT FROM AUTHOR]

Glutathione S-transferases (GSTs) are members of a multifunctional enzyme super family that plays a pivotal role in both insecticide resistance and protection against oxidative stress. In this study, we identified a single-copy gene, AccGSTD, as being a Delta class GST in the Chinese honey bee ( Apis cerana cerana). A predicted antioxidant response element, CREB, was found in the 1,492-bp 5′-flanking region, suggesting that AccGSTD may be involved in oxidative stress response pathways. Real-time PCR and immunolocalization studies demonstrated that AccGSTD exhibited both developmental- and tissue-specific expression patterns. During development, AccGSTD transcript was increased in adults. The AccGSTD expression level was the highest in the honey bee brain. Thermal stress experiments demonstrated that AccGSTD could be significantly upregulated by temperature changes in a time-dependent manner. It is hypothesized that high expression levels might be due to the increased levels of oxidative stress caused by the temperature challenges. Additionally, functional assays of the recombinant AccGSTD protein revealed that AccGSTD has the capability to protect DNA from oxidative damage. Taken together, these data suggest that AccGSTD may be responsible for antioxidant defense in adult honey bees. [ABSTRACT FROM AUTHOR]

Small heat shock proteins (sHSPs) play an important role in the cellular defense of prokaryotic and eukaryotic organisms against a variety of internal and external stressors. In this study, a cDNA clone encoding a member of the α-crystallin/sHSP family, termed AccHsp27.6, was isolated from Apis cerana cerana. The full-length cDNA is 1,014 bp in length and contains a 708-bp open reading frame encoding a protein of 236 amino acids with a calculated molecular weight of 27.6 kDa and an isoelectric point of 7.53. Seven putative heat shock elements and three NF-κB binding sites were present in the 5′-flanking region, suggesting a possible function in immunity. A semi-quantitative RT-PCR analysis indicated that AccHsp27.6 was expressed in all tested tissues and at different developmental stages. Furthermore, expression of the AccHsp27.6 transcript was induced by exposure to heat shock, HO, a number of different chemicals (including SO, formaldehyde, alcohol, acetone, chloroform, and the pesticides phoxime and acetamiprid), and the microbes Staphylococcus aureus and Micrococcus luteus. In contrast, the mRNA expression could be repressed by CO, the pesticides pyriproxyfen and cyhalothrin, and the microbes Bacillus subtilis and Pseudomonas aeruginosa. Notably, the recombinant AccHsp27.6 protein exhibited significant in vitro molecular chaperone activity and antimicrobial activity. Taken together, these results suggest that AccHsp27.6 might play an important role in the response to abiotic and biotic stresses and in immune reactions. [ABSTRACT FROM AUTHOR]

Calcyclin binding protein (CacyBP), a homolog of Sgt1, was shown to interact with some S100 proteins, Skp1, tubulin, actin and ERK1/2 kinases. Studies have also shown that CacyBP is a neuronal protein in mammals. Limited information is available regarding the properties and functions of CacyBP in insects. Here, we cloned and characterized a novel CacyBP gene, named AccCacyBP, from honeybee ( Apis cerana cerana). Bioinformatic analysis indicated that AccCacyBP was highly conserved and closely related to the CacyBP of other insects. Promoter analysis revealed a number of putative tissue, development and stress-related transcription factor-binding sites. RT-qPCR demonstrated that AccCacyBP was expressed at all of the stages of development, especially in the brains of honeybees. Moreover, immunohistochemistry analysis showed the presence of AccCacyBP in the brain. The transcript levels of AccCacyBP in the brains of honeybees were developmentally induced and upregulated by exposure to oxidative stresses, including UV-light, acetamiprid and HgCl. This study demonstrates that the CacyBP gene in honeybees may be a neuronal protein involved in the developmental regulation and the stress-response of the brain of honeybees. [ABSTRACT FROM AUTHOR]

Ribosomal proteins (RPs) play pivotal roles in developmental regulation. The loss or mutation of ribosomal protein L11 ( RPL11) induces various developmental defects. However, few RPs have been functionally characterized in Apis cerana cerana. In this study, we isolated a single copy gene, AccRPL11, and characterized its connection to brain maturation. AccRPL11 expression was highly concentrated in the adult brain and was significantly induced by abiotic stresses such as pesticides and heavy metals. Immunofluorescence assays demonstrated that AccRPL11 was localized to the medulla, lobula and surrounding tissues of esophagus in the brain. The post-transcriptional knockdown of AccRPL11 gene expression resulted in a severe decrease in adult brain than in other tissues. The expression levels of other brain development-related genes, p38, ERK2, CacyBP and CREB, were also reduced. Immunofluorescence signal attenuation was also observed in AccRPL11-rich regions of the brain in ds AccRPL11-injected honeybees. Taken together, these results suggest that AccRPL11 may be functional in brain maturation in honeybee adults. [ABSTRACT FROM AUTHOR]

Lipid storage droplet 1 (LSD-1), a PAT family protein located around lipid droplets in insects, is intimately linked to lipid droplets formation and lipid metabolism. Conjugated linoleic acid (CLA) and rosiglitazone (Rosi) have previously been shown to modulate the expression of several PAT family proteins through peroxisome proliferator-activated receptor-γ (PPARγ). In the present study, we isolated and characterized a novel LSD- 1 gene, referred to AccLSD- 1, from Chinese honeybee ( Apis cerana cerana). Sequence analysis indicated that the central region of LSD-1 protein had significant sequence similarity and a typical LSD- 1 gene was composed of 8 exons and 7 introns. Interestingly, the first intron of AccLSD- 1 including several PPARγ-response elements (PPREs) was located in 5′ UTR. Analysis of 5′-flanking region of AccLSD- 1 revealed a number of putative cis-acting elements, including three PPREs. Quantitative real-time PCR showed that AccLSD- 1 expressed ubiquitously from feeding larva to adult, and its expression level was highest at brown-eyed pupae (Pb) stage. The effect of CLA, Rosi and combination on AccLSD- 1 expressions indicated 1% CLA and 0.5 mg/ml Rosi were considered as the suitable diets for rearing adult workers in laboratory, and AccLSD- 1 was down-regulated by CLA whereas up-regulated by Rosi. Furthermore, the combination of CLA and Rosi remarkly rescued the suppression of AccLSD- 1 expression by CLA alone. These results suggest that AccLSD- 1 is associated with A. cerana cerana development, especially during pupal metamorphosis, and can be regulated by CLA or Rosi possibly via activating PPARγ. [ABSTRACT FROM AUTHOR]