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Liu Y.,ShenYang Agricultural University | Liu Y.,Sericultural Institute of Liaoning Province | Chen M.,Sericultural Institute of Liaoning Province | Su J.,Guangzhou University of Chinese Medicine | And 5 more authors.
PLoS ONE | Year: 2015

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. © 2015 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Chen M.-M.,ShenYang Agricultural University | Li Y.,ShenYang Agricultural University | Chen M.,ShenYang Agricultural University | Wang H.,ShenYang Agricultural University | And 7 more authors.
Gene | Year: 2014

Mitochondrial genome (mitogenome) can provide information for genomic structure as well as for phylogenetic analysis and evolutionary biology. In this study, we present the complete mitogenome of the atlas moth, Attacus atlas (Lepidoptera: Saturniidae), a well-known silk-producing and ornamental insect with the largest wing surface area of all moths. The mitogenome of A. atlas is a circular molecule of 15,282. bp long, and its nucleotide composition shows heavily biased towards As and Ts, accounting for 79.30%. This genome comprises 13 protein-coding genes (PCGs), two ribosomal RNA genes (rRNAs), 22 transfer RNA genes (tRNAs), and an A. +. T-rich region. It is of note that this genome exhibits a slightly positive AT skew, which is different from the other known Saturniidae species. All PCGs are initiated by ATN codons, except for COI with CGA instead. Only six PCGs use a common stop codon of TAA or TAG, whereas the remaining seven use an incomplete termination codon T or TA. All tRNAs have the typical clover-leaf structure, with an exception for tRNASer(AGN). The A. atlas A. +. T-rich region contains non-repetitive sequences, but harbors several features common to the Bombycoidea insects. The phylogenetic relationships based on Maximum Likelihood method provide a well-supported outline of Saturniidae, which is in accordance with the traditional morphological classification and recent molecular works. © 2014 Elsevier B.V.


Sun S.-H.,ShenYang Agricultural University | Li Y.-P.,ShenYang Agricultural University | Zheng Y.-N.,ShenYang Agricultural University | Xu X.-R.,ShenYang Agricultural University | And 5 more authors.
Annals of the Entomological Society of America | Year: 2011

Selenophosphate synthetase (Sps), the product of the SelD gene, produces a biologically active selenium donor compound from ATP and selenide. We have isolated and characterized the Sps gene from Antheraea pernyi (Guérin-Méneville) (Lepidoptera: Saturniidae), an economically important insect. The resulting 1601 bp cDNA sequence contains an open reading frame of 1209 bp encoding a polypeptide of 402 amino acids, with 87% sequence identity to that from Drosophila melanogaster (Meigen). Semiquantitative reverse transcription-polymerase chain reaction (PCR) analysis showed that the Sps gene was transcribed during four developmental stages (egg, larva, pupa, and adult) and in all the tissues tested (blood, fat body, midgut, silk glands, body wall, spermaries and ovaries), suggesting that ApSps plays an important role in the development of A. pernyi. From a database search, Sps protein homologs were found in prokaryotes and eukaryotes, including bacteria, fungi, invertebrates and vertebrates, with 47-98% amino acid sequence identities between eukaryotes, suggesting that they were highly conserved during the evolution of eukaryotes. Phylogenetic analysis, based on Sps protein homolog sequences, clearly separated the known bacterial, fungal, invertebrate and vertebrate Sps proteins, consistent with the topology tree of classical systematics, suggesting the potential value of the Sps protein sequence in phylogenetic inference. © 2011 Entomological Society of America.


Liu Y.,ShenYang Agricultural University | Li Y.,ShenYang Agricultural University | Li X.,Sericultural Institute of Liaoning Province | Qin L.,ShenYang Agricultural University
Journal of Insect Science | Year: 2010

Sericulture is one of the great inventions of the ancient Chinese. Besides the mulberry silkworm (Bombyx mori), Chinese farmers developed rearing of the Chinese oak silkworm (Antheraea pernyi) about 400 years ago. In this paper, the historic records of the origins and dispersal of the domesticated Chinese oak silkworm in China are summarized. The first document with clearly recorded oak silkworm artificial rearing appeared in 1651, although Chinese oak silkworm was documented in about 270 AD. All of the evidence in the available historic records suggests that the domesticated Chinese oak silkworm originated in central and southern areas of Shandong Province in China around the 16th century, and then was introduced directly and indirectly by human commerce into the present habitations in China after the late 17th century. The results strongly support the hypothesis that only one geographically distinct event occurred in domestication of the modern Chinese oak silkworm.


Liu L.,ShenYang Agricultural University | Wang H.-Y.,ShenYang Agricultural University | Jin H.-Y.,ShenYang Agricultural University | Wu S.,ShenYang Agricultural University | And 6 more authors.
Biochemical Systematics and Ecology | Year: 2010

Myosin light chain 2 (MLC-2) gene was isolated and characterized from Antheraea pernyi, a well-known wild silkmoth. The isolated cDNA sequence is 905 bp in length with an open reading frame of 612 bp encoding a polypeptide of 203 amino acids. Semi-quantitative RT-PCR analysis showed that the MLC-2 gene was transcribed during four developmental stages (egg, larva, pupa, and moth), and present in all tissues tested. Alignment analysis revealed that the deduced protein sequence has over 95% identity to myosin light chain 2 of lepidopteran species, and 57-88% identity to other insect species, suggesting that insect MLC-2 proteins are highly conserved throughout evolution. The protein sequence was used to construct phylogenetic trees with other known vertebrate and invertebrate MLC-2 sequences, and the obtained trees demonstrated similar topology with the classical systematics, indicating the potential value of MLC-2 gene in phylogenetic study. © 2010 Elsevier Ltd.


Jiang D.-F.,Sericultural Institute of Liaoning Province | Jiang D.-F.,ShenYang Agricultural University | Liu Y.-Q.,Sericultural Institute of Liaoning Province | Liu Y.-Q.,ShenYang Agricultural University | And 2 more authors.
African Journal of Biotechnology | Year: 2010

It has been known that the abnormal wing disc (awd) gene encodes a nucleoside diphosphate kinase and is closely related to wing development in Drosophila melanogaster and Bombyx mori. In the present study, the awd gene was isolated and characterized from Antheraea pernyi, a well-known wild silkmoth. The isolated cDNA sequence is 666 bp in length with an open reading frame of 462 bp encoding a polypeptide of 153 amino acids, which contains a putative nucleoside diphosphate kinases active site motif and conserved multimer interface. The deduced A. pernyi awd protein sequence reveals 75, 82 and 96% identity with its homologue of Homo sapiens, D. melanogaster, and B. mori, respectively. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that the awd gene was transcribed during all four developmental stages (egg, larva, pupa, and moth), and present in all tissues tested (blood, midgut, silk glands, Malpighian tublues, spermaries, ovaries, brain, muscle, fat body and body wall), with the highest abundance in Malpighian tubules. Interestingly, mRNA expression level in pupal fat body was significantly down-regulated after cold shock (4°C) compared with the control (26°C) and significantly up-regulated after heat shock (46°C). The results indicated that the A. pernyi awd gene is inducible, and that its expression effect is different after cold stress and heat stress. Consequently, we refer that the product of the awd gene may contribute to its temperature tolerance. © 2010 Academic Journals.


Chen M.,ShenYang Agricultural University | Chen M.-M.,ShenYang Agricultural University | Yao R.,ShenYang Agricultural University | Li Y.,ShenYang Agricultural University | And 4 more authors.
Journal of Agricultural and Food Chemistry | Year: 2013

Two 12 kDa FK506-binding protein (FKBP12) genes were isolated and characterized from Chinese oak silkworm Antheraea pernyi, an important agricultural and edible insect, designated ApFKBP12 A and B, respectively. Both ApFKBP12 A and B contained 108 amino acids with 82% sequence identity. Phylogenetic analysis showed that FKBP12 B sequences of A. pernyi, Bombyx mori, and Danaus plexippus were clearly separated from FKBP12 A sequences of these three species, suggesting that insect FKBP12 A and B may have been evolving independently. RT-PCR analyses revealed that two ApFKBP12 genes were expressed during the four developmental stages and in all tested tissues, and that the mRNA expression level of the ApFKBP12 A gene was significantly higher than that of the ApFKBP12 B gene. After heat shock treatment, expressions of the two FKBP12 genes were up-regulated, but at different time points. The results suggested that each paralogue of the FKBP12 genes may play a distinct functional role in the development of A. pernyi. © 2013 American Chemical Society.


Liu Y.,ShenYang Agricultural University | Liu Y.,Sericultural Institute of Liaoning Province | Li Y.,ShenYang Agricultural University | Wang H.,ShenYang Agricultural University | And 8 more authors.
Acta Biochimica et Biophysica Sinica | Year: 2010

In this study, two enolase genes were isolated and characterized from the Chinese oak silkworm, Antheraea perny, which were designated as enolase I and II, respectively. The enolase I cDNA sequence was 1712 bp with an open reading frame (ORF) of 1302 bp encoding 433 amino acids. The enolase II cDNA sequence was 1549 bp with an ORF of 1296 bp encoding 431 amino acids. The amino acid sequences of the two genes share several conserved features/sites of enolase. Antheraea pernyi enolase I shows 93-97 sequence identity to enolases of lepidopterans available to date, 75-82 identity to enolases of other invertebrates, 60-72 identity to enolases of other organisms including vertebrates, plants, and fungi. Antheraea pernyi enolase II shows 84 identity to Bombyx mori enolase II, but 60 identity to A. pernyi enolase I. In the phylogenetic tree, enolase II sequences from A. pernyi and B. mori were clearly separated from the majority of enolase sequences of higher organisms including A. pernyi and B. mori enolase I sequences. By sequence comparisons and phylogenetic analysis, we suggest that enolase II from A. pernyi and B. mori may be a new member of the enolase superfamily. Antheraea pernyi enolase I mRNA was found in all tested tissues whereas enolase II mRNA was expressed specifically in the spermaries and ovaries, suggesting that the product of enolase II gene may be related to reproduction. The transcript abundance of A. pernyi enolase I gene was significantly down-regulated after cold shock and significantly up-regulated after heat shock, suggesting that A. pernyi enolase I gene may be inducible by temperature stress. © The Author 2010.


Sima Y.-H.,Soochow University of China | Chen M.,ShenYang Agricultural University | Yao R.,ShenYang Agricultural University | Li Y.-P.,ShenYang Agricultural University | And 7 more authors.
Gene | Year: 2013

The complete mitochondrial genome (mitogenome) of the Ailanthus silkmoth, Samia cynthia cynthia (Lepidoptera: Saturniidae) was determined. The circular genome is 15,345bp long, and presents a typical gene organization and order for sequenced mitogenomes of Bombycidea species. The nucleotide composition of the genome is highly A+T biased, accounting for 79.86%. The AT skew of the genome is slightly negative, indicating the occurrence of more Ts than As, as found in other Saturniidae species. All protein-coding genes (PCGs) are initiated by ATN codons, except for COI and COII, which are tentatively designated by CGA and GTG, respectively, as observed in other insects. Four of 13 PCGs, including COI, COII, ATP6, and ND3, harbor the incomplete termination codons, T or TA. With an exception for tRNASer(AGN), all other tRNAs can form a typical clover-leaf structure of mitochondrial tRNA. The 359bp A+T-rich region of S. c. cynthia contains non-repetitive sequences, but harbors several features common to the Bombycidea insects, including the motif ATAGA followed by a poly-T stretch of 19bp, a microsatellite-like (AT)7 element preceded by the ATTTA motif, and a poly-A element upstream tRNAMet. The phylogenetic analyses support the morphology-based current hypothesis that Bombycidae and Saturniidae are monophyletic. Our result confirms that Saturniini and Attacini form a reciprocal monophyletic group within Saturniidae. © 2013 Elsevier B.V.


Liu Y.-Q.,Soochow University of China | Liu Y.-Q.,ShenYang Agricultural University | Chen M.-M.,ShenYang Agricultural University | Li Q.,ShenYang Agricultural University | And 9 more authors.
Annals of the Entomological Society of America | Year: 2012

KK-42 is an imidazole insect growth regulator. A (KK-42)-binding protein (KK-42BP) has been shown to be associated with diapause termination in pharate first instars of Antheraea yamamai (Gurin-Mneville) (Lepidoptera: Saturniidae). In this study, the (KK-42)-binding protein gene (ApKK-42BP) was characterized from Antheraea pernyi (Gurin-Mneville) (Lepidoptera: Saturniidae) undergoing a winter diapause as a pupa. Homologous comparison revealed that KK-42BPs from A. pernyi and A. yamamai were closely related to the known minor yolk proteins from the lepidopterans Bombyx mori L., Plodia interpunctella (Hbner), and Galleria mellonella L. The two KK-42BPs also contained a lipase-like region, as observed in the known lepidopteran yolk proteins. Phylogenetic analysis suggested that KK-42BP is a new member of the minor yolk proteins. Reverse transcription- polymerase chain reaction analyses showed that ApKK-42BP mRNA was expressed in all of the tissues tested, throughout four developmental stages, and in both female and male. Both in the brain and hemolymph, expression of ApKK-42BP mRNA level was similar between nondiapause-destined and diapause-destined larval individuals. ApKK-42BP mRNA was expressed in the predia-pause period of diapause-destined pupae, disappeared in early diapause and diapause stage, and reappeared in the postdiapause stage. These expression patterns stated that a lack of KK-42BP is associated with pupal diapause and its expression may be critical to nondiapausing individuals. Our results suggested that the KK-42BP gene is likely involved in a function other than as a yolk protein. © 2012 Entomological Society of America.

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