Chinese Oak Silkworm Antheraea Pernyi Research Articles

Comparative demographics, population projections and egg maturation patterns of four eupelmid egg parasitoids on the factitious host Antherae pernyi.

The genera Anastatus and Mesocomys (both Hymenoptera: Eupelmidae) are important solitary egg endoparasitoids as biological control agents for lepidopterous and hemipterous pests worldwide. Here, we comparatively evaluated the demographic parameters of four important eupelmid egg parasitoids (Anastatus fulloi, A. japonicus, Mesocomys albitarsis and M. trabalae) reared on the factitious host eggs of the Chinese oak silkworm Antheraea pernyi, using age-stage two-sex life tables, their population projections as well as egg maturation patterns. Both the age-specific net reproductive rate (lx mx ) and reproductive value (vxj ) increased initially and then gradually decreased with increasing age in all four parasitoid species. Overall, the two Mesocomys species had higher survival rates at stable age-stage distribution, peak reproductive values, and intrinsic rates of increase than the two Anastatus species. Mesocomys albitarsis had the longest longevity while A. japonicus had the longest oviposition days and mean generation time. The two Mesocomys species are thus projected to have faster population increase than the two Anastatus species. Adult females of all four parasitoid species emerged with only a few mature eggs (< 6 eggs) and most of their eggs were matured post-emergence (i.e., strict synovigeny). The estimated 90% of lifetime complement of reproduction (offspring) and realized days were 374 and 32 for A. japonicus, 337 and 22 for M. trabalae, 330 and 19 for M. albitarsis and 147 and 28 for A. fulloi. Our results indicated that the two Mesocomys species have higher control capacity than the two Anastatus species. Provision of adult food for these strictly synovigenic parasitoids would be essential to prolong their lifespan and continuously produce eggs for parasitizing their hosts for mass rearing or augmentative biological control programs. This article is protected by copyright. All rights reserved.

Evidence of two-sex life table analysis supporting Anastatus japonicus, a more effective biological control agent of Caligula japonica compared with other two Anastatus species

Japanese giant silkworm (JGS) Caligula japonica, is an emerging defoliator pest of forest and fruit trees, causing serious economic losses to walnut, ginkgo and chestnut in East Asia. To identify the dominant parasitoids of JGS, we compared biological control potential and fitness of three Anastatus species (A. fulloi, A. meilingensis and A. japonicus), which are natural egg parasitoids of JGS collected in northwestern China, on JGS eggs using age-stage, two-sex life table method. The results showed that three Anastatus species reared on suitable factitious host Chinese oak silkworm Antheraea pernyi were readily parasitized and completed their development and emergence from target JGS eggs. Anastatus japonicus had higher fecundity (F: 189.8 offspring/female) and longer oviposition days (Od: 24.36 d) than that of A. fulloi (F: 66.7 offspring/female, Od: 12.32 d) and A. meilingensis (F: 39.9 offspring/female, Od: 8.61 d). Consequently, the intrinsic rate of increase (r), finite rate of increase (λ) and net reproductive rate (R0) of A. japonicus (r: 0.1404 d−1, λ: 1.1508 d−1, R0: 146.5 offspring/individual) were significantly higher than those of A. fulloi (r: 0.1122 d−1, λ: 1.1187 d−1, R0: 50.9 offspring/individual) and A. meilingensis (r: 0.1213 d−1, λ: 1.1289 d−1, R0: 35.2 offspring/individual). In addition, the net killing rate (C0) of A. japonicus was 155.6 (eggs/female), which was significantly higher than that of A. fulloi (51.5 eggs/female) and A. meilingensis (36.7 eggs/female). Our results showed that A. japonicus had the highest parasitism capacity among three Anastatus species tested on target eggs and suggest that it may be the most suitable biocontrol agent for JGS.

Antheraea pernyi Suppressor of Cytokine Signaling 2 Negatively Modulates the JAK/STAT Pathway to Attenuate Microbial Infection.

The Janus kinase (JAK) signal transducer and activator of transcription (STAT) pathway has been shown to govern various physiological processes, including immune responses, hematopoiesis, cell growth, and differentiation. Recent studies show that suppressors of cytokine signaling (SOCS) proteins attenuate JAK-STAT signaling in mammals; however, their functions are less clear in lepidopteran insects. Here, we report a full-length sequence of SOCS-2 from the Chinese oak silkworm Antheraea pernyi (designated as ApSOCS-2) and study its biological role in immune responses via the JAK-STAT pathway. ApSOCS-2 expression was high in the fat bodies and hemocytes of A. pernyi fifth instar larvae. After pathogen infection with nucleopolyhedrovirus, Beauveria bassiana, Escherichia coli, and Microccus luteus, ApSOCS-2 mRNA was strongly increased compared to the control group. To elucidate the possible involvement in innate immunity, we measured antimicrobial peptide genes expression profiles in the fat body of A. pernyi. In contrast, recombinant ApSOCS-2 protein administration significantly reduced the AMPs transcription, while the depletion of ApSOCS-2 by RNAi increased their expression. Furthermore, we observed higher antibacterial activity and lower bacterial replication in dsApSOCS-2-treated larvae. The ApSOCS-2 transcription level was reduced in STAT depleted A. pernyi larvae challenged by M. luteus. The ApSOCS-2 RNAi data sets were also subjected to transcriptomic analysis, which suggests that ApSOCS-2 is a key regulator of immune function. Taken together, our data suggest that ApSOCS-2 is required for the negative regulation of AMPs transcripts via the JAK-STAT pathway in the insect.

Demography and Fitness of Anastatus japonicus Reared from Antheraea pernyi as a Biological Control Agent of Caligula japonica.

Simple SummaryJapanese giant silkworm (JGS), Caligula japonica Moore, is an emerging defoliator pest of forest and fruit trees in East Asia, causing severe economic losses. In this study, we used eggs of the Chinese oak silkworm (COS), Antheraea pernyi Guérin-Méneville, as an alternative host to rear the dominant JGS egg parasitoid, Anastatus japonicus Ashmead. We showed that A. japonicus could be efficiently reared on COS eggs. Furthermore, A. japonicus reared from COS eggs had a high biological control potential against JGS. This study provides useful information on the demography, parasitism rate, and rearing methods of A. japonicus on COS eggs and the basis for developing a cost-effective biological control program against JGS using A. japonicus.Japanese giant silkworm (JGS), Caligula japonica Moore, is an emerging defoliator pest of forest and fruit trees in East Asia, causing severe economic losses. To develop a cost-effective biological control program against JGS, we used eggs of the Chinese oak silkworm (COS) Antheraea pernyi Guérin-Méneville as an alternative host to rear the most dominant JGS egg parasitoid Anastatus japonicus Ashmead. We compared the demographic parameters and total parasitism (killing) rates of A. japonicus parasitizing JGS and COS eggs using an age-stage, two-sex life table method. The results showed that A. japonicus performed differently on these two different hosts. Anastatus japonicus reared from COS eggs had a higher fecundity (369.7 eggs per female) and a longer oviposition period (35.9 days) on the COS than JGS eggs (180.9 eggs; 24.0 days). Consequently, A. japonicus parasitizing COS eggs had a higher intrinsic rate of increase (r = 0.1466 d−1), finite rate of increase (λ = 1.1579 d−1) and net reproductive rate (R0 = 284.9 offspring) than those parasitizing JGS eggs (r = 0.1419 d−1, λ = 1.1525 d−1, R0 = 150.0 offspring). The total net parasitism rate (the number of parasitized hosts in which the parasitoids successfully developed) of A. japonicus parasitizing COS eggs was 284.9, significantly higher than that of A. japonicus parasitizing JGS eggs (150.0), while the net non-effective parasitism rate (the number of parasitized hosts in which the parasitoids failed to develop) of the former (0.0) was significantly lower than that of the latter (9.6). These results suggest that A. japonicus can be efficiently reared on the alternative (or factitious) COS eggs, and the reared parasitoids have a high biological control potential against the target JGS.

Chinese oak silkworm Antherae pernyi egg, a suitable factitious host for rearing eupelmid egg parasitoids.

Eupelmid egg parasitoids in the genera Anastatus and Mesocomys are important biological control agents for lepidopterous and hemipterous pests worldwide. The egg of Chinese oak silkworm Antheraea pernyi has been widely used for mass rearing of Trichogramma parasitoids. This study evaluated the suitability and optimal use methods of A. pernyi egg as a factitious host for the rearing of six eupelmid egg parasitoids (Anastatus fulloi, Anastatus gansuensis, Anastatus japonicus, Anastatus meilingensis, Mesocomys albitarsis and Mesocomys trabalae). Each parasitoid was tested for its oviposition preference and offspring performance on various differently treated host eggs (extracted from virgin moths or laid naturally by virgin or mated moths, and washed or unwashed prior to the use) in both no-choice and choice tests. All treated A. pernyi eggs were readily parasitized by the six parasitoids. In general, A. gansuensis and M. trabalae preferred washed over unwashed eggs regardless of the fertilization status of host eggs, A. fulloi and A. meilingensis parasitized more unfertilized than fertilized host eggs, and A. japonicus and M. albitarsis did not show a preference among differently treated host eggs. Host egg treatment did not significantly affect offspring fitness (development time, survival, sex ratio and body size) nor reproductive potential of developed adult females for each parasitoid species, except for M. albitarsis (whose females contained more eggs when reared from unfertilized than fertilized host eggs). Results suggest that manually extracted, unfertilized and washed A. pernyi eggs are most suitable for mass rearing of these eupelmid egg parasitoids in biological control programs.

NISES-AnPe-428 cell line derived from the Chinese oak silkworm Antheraea pernyi is permissive for multiple nucleopolyhedrovirus species from insects of four different families.

The cell line NISES-AnPe-428 (AnPe), derived from the Chinese oak silkworm Antheraea pernyi, was characterized for its permissiveness and productivity for six different nucleopolyhedrovirus (NPV) species. These NPVs included homologous Antheraea pernyiNPV (AnpeNPV) and heterologous Autographa californicamultiple NPV (AcMNPV), Bombyx moriNPV (BmNPV), Hyphantria cuneaMNPV (HycuMNPV), Spodoptera exiguaMNPV (SeMNPV), and Lymantria disparMNPV (LdMNPV), representing viruses that had been isolated from insect species belonging to five different families (Saturniidae, Noctuidae, Bombycidae, Arctiidae, and Lymantriidae). We found that AnPe cells supported productive replication of AnpeNPV, AcMNPV, BmNPV, HycuMNPV, and SeMNPV to varying degrees. Upon infection with SeMNPV, a subset of AnPe cell population in the culture underwent apoptosis, while remaining cells produced limited amounts of progeny virions and polyhedra. AnPe cells were refractory to LdMNPV infection and failed to support replication of viral DNA, indicating that viral replication was restricted at or prior to the step of viral DNA replication. These results indicated that AnPe cells have the potential to provide excellent systems for studying the molecular mechanisms underlying cellular permissiveness for NPV replication and host-range determination of NPVs.

Comparison of bacterial communities between midgut and midgut contents in two silkworms, Antheraea pernyi and Bombyx mori

Bacterial communities living inside the midgut of insects have been attracting increasing interest. Previous studies have shown that both the midgut and midgut contents harbor bacterial communities. However, whether the bacterial communities of the insect midgut are similar to those of the insect midgut contents (including the peritrophic membrane, food particles, and digestive fluids secreted by the midgut in this study) remains unknown. In the present study, we analyzed two economically important silkworms, the Chinese oak silkworm Antheraea pernyi (Lepidoptera: Saturniidae) and the mulberry silkworm Bombyx mori (Lepidoptera: Bombycidae), through Illumina MiSeq technology to address this issue. In A. pernyi larvae, 17 phyla and 162 genera were found in the midgut, while 7 phyla and 36 genera were found in the midgut contents. For B. mori larvae, 30 phyla and 465 genera were found in the midgut, but 22 phyla and 344 genera were found in the midgut contents. This evidence from the two silkworms suggests that the bacterial composition and diversity in the midgut are more diverse than those in the midgut contents. Principal component analysis revealed a significant difference in the bacterial community structure between the midgut and midgut contents of B. mori. To our knowledge, this is the first study to compare the bacterial communities between the midgut and midgut contents in insects, and the results will provide useful information for probing the functional differentiation within the midgut in the future.

In vitro rearing of Ooencyrtus nezarae (Hymenoptera: Encyrtidae), an egg parasitoid of Riptortus pedestris (Hemiptera: Alydidae)

ABSTRACTOoencyrtus nezarae Ishii (Hymenoptera: Encyrtidae) is a polyphagous egg parasitoid of various true bugs, including Riptortus pedestris (Fabricius) (Hemiptera: Alydidae), a major pest of soybean and fruit trees in northeastern Asia. This study was conducted to develop artificial host eggs containing insect haemolymph for mass rearing of O. nezarae. The haemolymph of Chinese oak silkworm (Antheraea pernyi Guérin-Méneville) pupae was found to be most suitable for artificial eggs for the complete development of O. nezarae. Among 764 parasitoid eggs laid in artificial eggs, 49.2% successfully developed to adult parasitoids. The developmental time in artificial eggs was delayed by 3–5 days relative to that observed in natural host eggs; however there was no significant difference in the length of hind tibia between adults that emerged from artificial and natural host eggs. Moreover, new generations that emerged from artificial eggs reproduced on all egg types offered (either natural or artificial eggs). We propose that the artificial eggs developed herein can therefore be an effective means for mass rearing O. nezarae.

A small heat shock protein 21 (sHSP21) mediates immune responses in Chinese oak silkworm Antheraea pernyi

Small heat shock proteins (sHSPs) are conserved among insects and play an important role in the regulation of many biological processes, including temperature stress, abiotic stress, immune responses, metamorphosis, and embryo development. Antheraea pernyi is an economically valuable silk-producing moth and source of insect food containing high-quality protein. The aim of this study was to quantify expression of the ApsHSP21 gene in response to pathogen-associated molecular patterns (PAMPs) and nucleopolyhedrovirus (NPV) challenge. The deduced ApsHSP21 protein sequence consists of 186 residues with a calculated molecular mass of 21.0 kDa and an isoelectronic point (pI) of 6.63. The protein contains a conserved α-crystallin domain (ACD), and includes two casein kinase II phosphorylation sites, a protein kinase C phosphorylation site, two tyrosine kinase phosphorylation sites, and various polypeptide binding sites. Phylogenetic analysis revealed that ApsHSP21 is closely related to homologs from other insects. Real-time quantitative reverse transcription PCR (qRT-PCR) analysis revealed that expression of ApsHSP21 was significantly up-regulated at different timepoints following simulated pathogen challenge with lipopolysaccharide (LPS), peptidoglycan (PGN), glucan, and NPV. The results suggest sHSP21 is involved in innate immune responses in A. pernyi.

Transcriptome-Wide Identification of Differentially Expressed Genes in Chinese Oak Silkworm Antheraea pernyi in Response to Lead Challenge.

Antheraea pernyi is a commercially cultivated silk moth and a source of insect food with high-quality protein. Insects suffer oxidative stress on exposure to heavy metals, and reactive oxygen species are cleared by antioxidant enzymes. To gain better understanding of the antioxidant defense system of A. pernyi, we analyzed transcriptomes of pupae after stimulation with lead and phosphate-buffered saline (control). In total, 72 367 unigenes were identified. Gene ontology analysis revealed that these DEGs were in 20 biological process subcategories, 19 cellular component subcategories, and 18 molecular function subcategories. Clusters of orthologous groups of protein annotation placed a total of 528 DEGs into 25 categories. Kyoto Encyclopedia of Genes and Genomes enrichment analysis identified antioxidant defense pathways, including "Peroxisome" and "Glutathione metabolism", which are reported for the first time in A. pernyi. Our study enriches A. pernyi transcriptome databases and provides insight into the heavy metal responses of antioxidant systems of this insect fat bodies.

Molecular cloning and expression analysis of the highly conserved eukaryotic translation initiation factor 5A (eIF‐5A) from Antheraea pernyi

AbstractEukaryotic initiation factor 5A (eIF‐5A) is a highly conserved protein found in all eukaryotic organisms that plays a key role in the regulation of many cellular processes including translation elongation, cell proliferation, programmed cell death, mRNA turnover and decay, and abiotic stress responses. In this study, the eIF‐5A gene from the Chinese oak silkworm Antheraea pernyi (Lepidoptera: Saturniidae) was characterized. The full‐length ApeIF‐5A cDNA of 1056 bp includes a 5′‐untranslated region (UTR) of 138 bp, a 3′‐UTR of 435 bp, and an open reading frame of 483 bp encoding a polypeptide of 160 amino acids. The deduced ApeIF‐5A protein shares 99 %, 82 %, and 72 % sequence identity with orthologs in Bombyx mori, Drosophila melanogaster and Homo sapiens, indicating high conservation during animal evolution. Real‐time quantitative reverse transcription PCR revealed expression in all four developmental stages and in all nine tissues tested, consistent with an important role in development. After challenge with lipopolysaccharide, the expression levels of ApeIF‐5A were markedly upregulated. Phylogenetic analysis of amino acid sequences revealed A. pernyi eIF‐5A was closely related to B. mori eIF‐5A, consistent with traditional classification and other molecular data. The results indicate the potential value of eIF‐5A in phylogenetic analysis.

Serpin-14 negatively regulates prophenoloxidase activation and expression of antimicrobial peptides in Chinese oak silkworm Antheraea pernyi

Genes encoding proteins of serpins superfamily are widely distributed in invertebrates. In insects, serpins play important roles in regulating immune responses and other physiological processes. Here, we report the cloning and characterization of cDNA of Apserpin-14 from Chinese oak silkworm (Antheraea pernyi). The Apserpin-14 gene contains 1206 bp open reading frame, encoding a predicted 401 amino acid residue protein. We expressed the recombinant Apserpin-14 protein in Escherichia coli and then purified protein was used to prepare rabbit anti-Apserpin-14 polyclonal antibodies. Quantitative real-time PCR analysis revealed that mRNA level of Apserpin-14 was highest in the fat body, whereas, among developmental stages the 5th instar and pupal stage showed greatest expression. Furthermore, Escherichia coli, Beauveria bassiana, Micrococcus luteus and nuclear polyhedrosis virus challenge enhanced Apserpin-14 transcript in both the fat body and hemocyte. Recombinant Apserpin-14 added to hemolymph inhibited spontaneous melanization and suppressed prophenoloxidase activation stimulated by M. luteus, but did not affect phenoloxidase (PO) activity. Injection of recombinant Apserpin-14 protein into A. pernyi larvae significantly reduced the transcript levels of antimicrobial peptides in the fat body, while its depletion by double stranded RNA enhanced their expression. We concluded that Apserpin-14 likely involved in regulation of proPO activation and production of antimicrobial peptides, implying its important role in the innate immune system of A. pernyi.

The functions of serpin-3, a negative-regulator involved in prophenoloxidase activation and antimicrobial peptides expression of Chinese oak silkworm, Antheraea pernyi

Serpins are a superfamily of proteins engaged in various physiological processes in all kingdoms of life. To date, many striking results have demonstrated serpins are involved in the invertebrate immune system by regulating the proteolytic cascades. However, in most insect species, the immune functions of serpins in response against pathogen invasion remain obscure. In this study, we identified a full-length cDNA sequence of serpin, named serpin-3, from the Chinese oak silkworm Antheraea pernyi. Sequence alignments have indicated that Apserpin-3 might regulate the melanization reaction via inhibiting prophenoloxidases-activating protease(s) in plasma. Furthermore, it was detected to be primarily transcribed within the fat body, epidermis and hemocytes with significant induction following immune-challenge. Further studies have shown that the knockdown of serpin-3 up-regulated the prophenoloxidases cascade stimulated by pathogen in hemolymph, while the addition of recombinant serpin-3 along with the same elicitor led to the suppressed activation of prophenoloxidase. Besides, the injection of dsRNA of serpin-3 caused the elevated expression of antimicrobial peptides. Altogether, we arrived at a conclusion that serpin-3 might act as a negative-regulator in prophenoloxidases activation and inhibit the production of antimicrobial peptides in Antheraea pernyi larvae.

Transcriptome Analysis of the Midgut of the Chinese Oak Silkworm Antheraea pernyi Infected with Antheraea pernyi Nucleopolyhedrovirus.

The Antheraea pernyi nucleopolyhedrovirus (ApNPV) is an exclusive pathogen of A. pernyi. The intense interactions between ApNPV and A. pernyi cause a series of physiological and pathological changes to A. pernyi. However, no detailed report exists regarding the molecular mechanisms underlying the interactions between ApNPV and A. pernyi. In this study, four cDNA libraries of the A. pernyi midgut, including two ApNPV-infected groups and two control groups, were constructed for transcriptomic analysis to provide new clues regarding the molecular mechanisms that underlie these interactions. The transcriptome of the A. pernyi midgut was de novo assembled using the Trinity platform because of the lack of a genome resource for A. pernyi. Compared with the controls, a total of 5,172 differentially expressed genes (DEGs) were identified, including 2,183 up-regulated and 2,989 down-regulated candidates, of which 2,965 and 911 DEGs were classified into different GO categories and KEGG pathways, respectively. The DEGs involved in A. pernyi innate immunity were classified into several categories, including heat-shock proteins, apoptosis-related proteins, serpins, serine proteases and cytochrome P450s. Our results suggested that these genes were related to the immune response of the A. pernyi midgut to ApNPV infection via their essential roles in regulating a variety of physiological processes. Our results may serve as a basis for future research not only on the molecular mechanisms of ApNPV invasion but also on the anti-ApNPV mechanism of A. pernyi.

The Toll Signaling Pathway in the Chinese Oak Silkworm, Antheraea pernyi: Innate Immune Responses to Different Microorganisms.

The Toll pathway is one of the most important signaling pathways regulating insect innate immunity. Spatzle is a key protein that functions as a Toll receptor ligand to trigger Toll-dependent expression of immunity-related genes. In this study, a novel spatzle gene (ApSPZ) from the Chinese oak silkworm Antheraea pernyi was identified. The ApSPZ cDNA is 1065 nucleotides with an open reading frame (ORF) of 777 bp encoding a protein of 258 amino acids. The protein has an estimated molecular weight of 29.71 kDa and an isoelectric point (PI) of 8.53. ApSPZ is a nuclear and secretory protein with no conserved domains or membrane helices and shares 40% amino acid identity with SPZ from Manduca sexta. Phylogenetic analysis indicated that ApSPZ might be a new member of the Spatzle type 1 family, which belongs to the Spatzle superfamily. The expression patterns of several genes involved in the Toll pathway were examined at different developmental stages and various tissues in 5th instar larvae. The examined targets included A. pernyi spatzle, GNBP, MyD88, Tolloid, cactus and dorsalA. The RT-PCR results showed that these genes were predominantly expressed in immune-responsive fat body tissue, indicating that the genes play a crucial role in A. pernyi innate immunity. Moreover, A. pernyi infection with the fungus Nosema pernyi and the gram-positive bacterium Enterococcus pernyi, but not the gram-negative bacterium Escherichia coli, activated the Toll signaling pathway. These results represent the first study of the Toll pathway in A. pernyi, which provides insight into the A. pernyi innate immune system.

Identification and Characterization of a Novel Microvitellogenin from the Chinese Oak Silkworm Antheraea pernyi.

Microvitellogenin (mVg) is a relatively small vitellogenic protein only characterized in the eggs of the lepidopteran insects Manduca sexta and Bombyx mori. In the present study, we report a novel mVg (ApmVg) isolated from the Chinese oak silkworm Antheraea pernyi. The obtained ApmVg cDNA sequence contains an open reading frame of 783 bp encoding a protein of 260 amino acids with a predicted molecular weight of 29.96 kDa. This gene does not contain introns. Structural analysis revealed that this protein shares putative conserved domains with the lepidopteran low-molecular weight lipoprotein, which belongs to the lipoprotein_11 superfamily. The protein sequence of ApmVg exhibits 48% sequence identity with mVg from M. sexta and 40–47% sequence identity with the 30K lipoproteins from B. mori. Phylogenetic analysis suggests that ApmVg is a novel member of the lepidopteran low-molecular weight lipoproteins. Transcriptional analysis indicated that ApmVg mRNA is mainly expressed in the fat body (both female and male) during post-diapause development of the pupal stage, and it was also detected in ovaries and spermaries in smaller amounts. RT-PCR and Western blot analyses revealed that ApmVg is synthesized by the fat body and secreted into hemolymph and ultimately accumulates in eggs. The ApmVg transcript can be detected in the fat bodies of female pupae four days after treatment with 20-hydroxyecdysone and shows an expression pattern distinct from that of vitellogenin (Vg), which is detectable throughout diapausing and in post-diapause development. ApmVg decreased dramatically during embryonic development. These results represent the first study of mVg outside M. sexta and B. mori and provide insight into the physiological role and evolution of mVgs.

Imidazole derivative KK-42 boosts pupal diapause incidence and delays diapause termination in several insect species

The imidazole derivative KK-42 is a synthetic insect growth regulator known previously to be capable of averting embryonic diapause in several Lepidoptera, but whether it also affects diapauses occurring in other developmental stages remains unknown. In the present study, we examined the effect of KK-42 on pupal diapause in two species of Lepidoptera, the Chinese oak silkworm Antheraea pernyi and the corn earworm Helicoverpa zea, and in one species of Diptera, the flesh fly Sarcophaga crassipalpis. In A. pernyi, KK-42 delayed pupal diapause termination under the long day conditions that normally break diapause in this species. Likewise, in H. zea, KK-42 delayed termination of pupal diapause, a diapause that, in this species, is normally broken by high temperature. KK-42-treated pupae of these two species eventually terminated diapause and successfully emerged as adults, but the timing of diapause termination was significantly delayed. KK-42 also significantly increased the incidence of pupal diapause in H. zea and S. crassipalpis when administered to larvae that were environmentally programmed for diapause, but it was not capable of inducing pupal diapause in H. zea if larvae were reared under environmental conditions that do not normally evoke the diapause response. Experiments with H. zea showed that the effect of KK-42 on pupal diapause was dose- and stage-dependent, but not temperature-dependent. Results presented here are consistent with a link between KK-42 and the ecdysteroid signaling pathway that regulates pupal diapause.

Eicosanoids mediate sHSP 20.8 gene response to biotic stress in larvae of the Chinese oak silkworm Antheraea pernyi

Small heat shock proteins (sHSPs) can regulate protein folding and protect cells from stress. To investigate the role of sHSPs in the silk-producing insect Antheraea pernyi (A. pernyi; Lepidoptera: Saturniidae), cDNA encoding HSP20.8 in A. pernyi, termed Ap-sHSP20.8, was identified as a 564bp ORF. The translated amino acid sequence encoded 187 residues with a calculated molecular mass of 20.8kDa and an isoelectronic point (pI) of 5.98; the sequence showed homology to sHSP chaperone proteins from other insects. Ap-sHSP20.8 mRNA transcript expression was abundant in the midgut and fat body and found to be both constitutive and inducible by infectious stimuli. Therefore, Ap-sHSP20.8 may play important roles in A. pernyi immune responses under biotic stress. Furthermore, we found that eicosanoids could mediate the induction of Ap-sHSP20.8 in the fat body and midgut. Our findings show that sHSPs may be promising molecules to target in order to cripple immunity in insect pests.

Solid artificial diets for the phytoseiid predator Amblyseius swirskii

Amblyseius swirskii (Athias-Henriot) (Acari: Phytoseiidae) is a key predator of a wide range of pests including thrips, whitefly and several mite pests. A number of artificial diets have been developed for this predator, but all of these are liquid, complicating their use in mass production. In the present study, we investigated the survival, development and reproduction of A. swirskii fed on several dry artificial diets: the tested diets were freeze dried forms of previously developed liquid meridic artificial diets supplemented with extracts of decapsulated cysts of Artemia franciscana Kellogg (Anostraca: Artemiidae) or with pupal hemolymph of Chinese oak silkworm Antheraea pernyi (Guerin-Meneville) (Lepidoptera: Saturniidae), and newly composed powdered meridic artificial diets supplemented with ground dry A. franciscana cysts or lyophilized pupal hemolymph of A. pernyi. Performance of the mite on the artificial diets was compared with that on cattail pollen (Typha latifolia L.). Developmental time of A. swirskii females offered lyophilized diets was significantly shorter than on powdered diets. Total fecundity was significantly higher for females fed on the lyophilized diets than for those maintained on the powdered diet with A. franciscana. Daily oviposition rates were similar on T. latifolia pollen and both lyophilized diets but lower on both powdered diets. The highest intrinsic rate of increase was observed when A. swirskii was fed on T. latifolia pollen (0.210 females per female per day), followed by the freeze dried diets enriched with A. pernyi and A. franciscana (0.195 and 0.184 females per female per day, respectively), and the lowest growth rates were observed on the powdered diets supplemented with A. franciscana and A. pernyi (0.159 and 0.158 females per female per day, respectively). In conclusion, the phytoseiid was able to effectively feed on solid, powdered artificial diets. Freeze-drying of liquid diets did not influence their value to support the development and reproduction of A. swirskii. For mass rearing purposes, these dry diets have several advantages over liquid ones, including more convenient application and storage. Furthermore, they are believed to have better potential for use as supplemental foods to sustain predatory mite populations in the crop after release.

Artificial and factitious foods support the development and reproduction of the predatory mite Amblyseius swirskii

The generalist predatory mite Amblyseius swirskii Athias-Henriot (Acari: Phytoseiidae) was reared on Ephestia kuehniella Zeller eggs (Lepidoptera: Pyralidae), decapsulated dry cysts of the brine shrimp Artemia franciscana Kellogg (Anostraca: Artemiidae), and on meridic artificial diets (composed of honey, sucrose, tryptone, yeast extract, and egg yolk) supplemented with pupal hemolymph of the Chinese oak silkworm Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae) (AD1), with E. kuehniella eggs (AD2) or with A. franciscana cysts (AD3). Development, reproduction and predation capacity of the predatory mites were assessed in the first (G1) and sixth generation (G6) of rearing on the different diets. Immature survival rates in G1 were similar on all diets (96.8-100 %). After six generations, however, survival of A. swirskii was significantly reduced on all diets except on A. franciscana cysts. Oviposition rates did not differ between generations when females were fed on E. kuehniella, AD2 or AD3. The total number of deposited eggs was similar among diets except in G6 where the females fed on A. franciscana cysts produced more eggs than those maintained on E. kuehniella eggs. On most diets the intrinsic rates of increase in G1 were superior to those in G6, except for predators supplied with A. franciscana cysts where no differences were observed among generations. Female mites did not lose their capacity to kill first instar Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) after six generations on the different diets, but predation rates in G6 on E. kuehniella were lower than in G1. In conclusion, the different factitious and artificial diets tested in the present study supported the development and reproduction of A. swirskii for a single generation but fitness losses occurred to a varying degree after several generations on E. kuehniella eggs or the artificial diets. Artificial diet enriched with A. franciscana cysts yielded better results than the other artificial diets. Amblyseius swirskii performed best on decapsulated Artemia cysts indicating their potential for use in the mass production of the predator or to sustain its populations in the crop after release.