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Detection of food source by PCR analysis of the gut contents of Aldrichina grahami （Aldrich）（Diptera： Calliphoridae） during post-feeding period

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Insect Science 2007年第1期14卷 47-52页

作者： KAI LI gong-yin ye JIA-yinG ZHU CUI HU Institute of Insect Sciences College of Agriculture and Biotechnology Zhejiang University Hangzhou China

The last meal of sarcophagous maggots may be useful in identifying the species on whose flesh they have fed （the＂host＂species）. The DNA profile of the host species may indeed be detectable in the＂last meal＂. In this paper, mitochondrial DNA analysis of gut contents was used to identify the prior host of post-feeding larvae of Aldrichina grahami （Aldrich）（Diptera： Calliphoridae）. A modified logistic equation was fitted to estimate the probability of identifying the host under five different constant temperatures （16, 20, 24, 28 and 32 ℃）. Our results shows that the detected time ranged from a maximum of 24 h at 32℃ to 42 h at 16℃ and a minimum of 12 h at 32~C to 30 h at 16℃. Furthermore, the host detection time was also calculated to give the maximal time after larval hatching from the egg. These results indicate that, in criminal cases where the maggots stray from the corpse, the last meal of the larvae should not be overlooked as potentially critical evidence.

关键词： Cytochrome b gene detectable time forensic entomology gut post-feeding

Proteome changes in the plasma of Papilio xuthus (Lepidoptera:Papilionidae):effect of parasitization by the endoparasitic wasp Pteromalus puparum (Hymenoptera:Pteromalidae)

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第6期10卷 445-453页

作者： Jia-ying ZHU gong-yin ye Qi FANG Cui HU State Key Laboratory of Rice Biology Institute of Insect Sciences Zhejiang University Hangzhou 310029 China

Although the biochemical dissection of parasitoid-host interactions is becoming well characterized, the molecular knowledge concerning them is minimal. In order to understand the molecular bases of the host immune response to parasitoid attack, we explored the response of Papilio xuthus parasitized by the endoparasitic wasp Pteromalus puparum using proteomic approach. By examining the differential expression of plasma proteins in the parasitized and unparasitized host pupae by two-dimensional (2D) electrophoresis, 16 proteins were found to vary in relation to parasitization compared with unparasitized control samples. All of them were submitted to identification by mass spectrometry coupled with a database search. The modulated proteins were found to fall into the following functional groups: humoral or cellular immunity, detoxification, energy metabolism, and others. This study contributes insights into the molecular mechanism of the relationships between parasitoids and their host insects.

关键词： Proteomics Plasma Parasitism Immune response Pteromaluspuparum Papilio xuthus

Proteome changes in the plasma of Pieris rapae parasitized by the endoparasitoid wasp Pteromalus puparum

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2011年第2期12卷 93-102页

作者： Jia-ying ZHU Qi FANG gong-yin ye Cui HU State Key Laboratory of Rice Biology and Key Laboratory of Molecular Biology of Crop Pathology and Insects of Ministry of Agriculture Institute of lnsect Sciences Zhejiang University Hangzhou 310029 China Key Laboratory of Forest Disaster Warning and Control of Yunnan Province and Key Laboratory of Southwest Mountain Forest Resources Conservation and Utilization of Ministry of Education Southwest Forestry University Kunming 650224 China

Parasitism by the endoparasitoid wasp Pteromalus puparum causes alterations in the plasma proteins of Pieris rapae. Analysis of plasma proteins using a proteomic approach showed that seven proteins were differentially expressed in the host pupae after 24-h parasitism. They were masquerade-like serine proteinase homolog (MSPH), enolase (Eno), bilin-binding protein (BBP), imaginal disc growth factor (IDGF), ornithine decarboxylase (ODC), cellular retinoic acid binding protein (CRABP), and one unknown function protein. The full length cDNA sequences of MSPH, Eno, and BBP were successfully cloned using rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis indicated that the transcript levels of MSPH and BBP in the fat bodies of host pupae were inducible in response to the parasitism and their variations were consistent with translational changes of these genes after parasitism, while the transcript levels of Eno and IDGF were not affected by parasitism. This study will contribute to the better understanding of the molecular bases of parasitoid-induced host alterations associated with innate immune responses, detoxification, and energy metabolism.

关键词： Proteomics Plasma Parasitism Gene cloning Pteromalus puparum Pieris rapae

Heat shock protein genes （hsp20, hsp75 and hsp90） from Pieris rapae： Molecular cloning and transcription in response to parasitization by Pteromalus puparum

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Insect Science 2013年第2期20卷 183-193页

作者： Jia-ying Zhu Guo-Xing Wu gong-yin ye Cui Hu Key Laboratory of Forest Disaster Warning and Control of Yunnan Province Southwest Forestry University Kunming College of Plant Protection Yunnan Agricultural University Kunming Key laboratory of Molecular Biology of Crop Pathology and Insects of Ministry of Agriculture Institute of Insect Sciences Zhejiang University Hangzhou China

Most molecular work on the roles of heat shock proteins （hsps） in host-parasite interaction has focused on vertebrates, rather than invertebrates. Here the full length complementary DNA （cDNA） sequences of three hsp genes （hsp20, hsp75 and hsp90） were amplified from Pieris rapae, and their transcriptional responsiveness to parasitization by the endoparasitic wasp Pteromalus puparum were investigated. The cDNA sequence analysis of hsp20, hsp75 and hsp90 revealed open reading frames of 531, 2328 and 2 157 bp in length, which encode proteins with calculated molecular weights of 19.5, 75.48 and 82.7 kDa, respectively. The comparison of amino acid sequences showed that P rapae hsp20 shared highly divergent homology to that of other insects, while hsp75 and hsp90 showed high homology to their counterparts of other species. The expression analysis indicated that these three genes were influenced in response to parasitization by P. puparum. The hsp20 transcripts in parasitized pupae were higher compared to non- parasitized pupae. The expression of hsp75 and hsp90 were down-regulated following parasitization. The results indicate that hsps are involved in host-parasitoid interactions.

关键词： gene expression heat shock protein parasitization

Parasitism of Pieris rapae （Lepidoptera： Pieridae） by the endoparasitic wasp Pteromalus puparum （Hymenoptera： Pteromalidae）： Effects of parasitism on differential hemocyte counts, micro- and ultra-structures of host hemocytes

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Insect Science 2012年第4期19卷 485-497页

作者： Qian-Qian Zhang Jia Huang Jia-ying Zhu gong-yin ye State Key Laboratory of Rice Biology&Key laboratory of Molecular Biology of Crop Pathology and Insects of Ministry of Agriculture Institute of Insect SciencesZhejiang UniversityHangzhouChina

Parasitism by the endoparasitic wasp Pteromalus puparum （Hymenoptera： Pteromalidae） by using only its associated venom, can suppress the immunal responses of Pieris rapae （Lepidoptera： Pieridae）. However, up to now, current knowledge of the mech- anisms has been limited. The response of host hemocytes to parasitism was investigated using a combination of light and transmission electron microscopy （TEM）. Five hemocyte types, prohemocytes （PRs）, granulocytes （GRs）, plasmatocytes （PLs）, oenocytoids （OEs） and coagulocytes （COs）, were observed and characterized from both unparasitized and parasitized Pieris rapae pupae. Light microscopy showed that both GRs and PLs became more round and spread abnormally after parasitism, whereas the shape of other types of hemocytes remained unaffected. In addition, the size of PRs and PLs became larger while OEs became smaller. The proportion of PRs significantly increased after parasitism and that of PLs decreased by 43.9%, but there was no significant increase of GRs and OEs. TEM showed that all types of hemocytes except COs were damaged to various degrees after parasitism, especially resulting in electron opaque cytoplasm and nucleus, fewer cell organelles of rough endoplasmic reticulum, mitochondria and vesicles. Our results indicate that parasitism by P. puparum affects differential hemocyte counts and structures of host hemocytes, particularly for GRs and PLs, which may be the main cause of the parasitoid suppressing host cellular immune responses.

关键词： Key words hemocytes light microscopy parasitism Pieris rapae Pteromaluspuparum,transmission electron microscopy ultrastructure

An endoparasitoid uses its egg surface proteins to regulate its host immune response

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Insect Science 2022年第4期29卷 1030-1046页

作者： Zi-Wen Teng Hui-Zi Wu Xin-Hai ye Qi Fang Hong-Xu Zhou gong-yin ye China-Australia Cooperation Base of Crop Health and Invasive Species China-Australia Joint Institute of Agricultural and Environmental HealthCollege of Plant Health and MedicineQingdao Agricultural UniversityQingdaoShandong ProvinceChina State Key Laboratory of Rice Biology Ministry of Agricultural and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and InsectsKey Laboratory of Biology of Crop Pathogens and Insects of Zhejiang ProvinceInstitute of Insect SciencesZhejiang UniversityHangzhouChina

With proteomic analysis,we identified 379 egg surface proteins from an endoparasitoid,Cotesia *** containing conserved enzymatic domains constitute a large proportion of egg surface *** proteins,such as superoxidase dismutase,homolog of *** 32-kDa protein,and immunoevasive protein-2A,are classical parasitism factors that have known functions in host immunity *** assays revealed that a novel egg surface protein,*** egg surface serpin domain-containing protein had the same function as a *** venom serpin,as both suppressed host melanization in a dose-dependent *** egg surface serpin domain-containing protein is mainly transcribed in *** oocytes with follicular cells,and it is located on both the anterior and posterior sides of the mature egg ***,we used LC-MS/MS to identify 586 binding proteins sourced from *** plasma located on the eggshell surface of ***,which included some immunity-related *** results not only indicate that *** uses its egg surface proteins to reduce the immune response of its host but also imply that endoparasitoid egg surface proteins might be a new parasitism factor involved in host immune regulation.

关键词： egg surface protein immune regulation melanization parasitism factor parasitoid wasp

cDNA of an arylphorin-type storage protein from Pieris rapae with parasitism inducible expression by the endoparasitoid wasp Pteromalus puparum

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Insect Science 2009年第3期16卷 227-236页

作者： Jia-ying Zhu gong-yin ye Qi Fang Cui Hu Zunnu-Raen Akhtar State Key Laboratory of Rice Biology Institute of Insect Sciences College of Agriculture and Biotechnology Zhejiang University Hangzhou China

This report presents the cDNA cloning of a storage protein, PraAry, from Pieris rapae and investigates its expression regulated by parasitism of an endoparasitoid wasp Pteromalus puparum. The full-length eDNA of PraAry is 2 270 nucleotides and contains a 2 121 nucleotide open reading frame encoding 707 amino acids with calculated molecular weights of approximately 83 kDa. Analysis of the primary protein sequence revealed that it possesses a signal peptide of 16 amino acids at the N-terminus and contains two highly conserved storage protein signature motifs. According to both phylogenetic analysis and the criteria for amino acid composition, PraAry belongs to the subfamily of arylphorin-type storage protein （1.42% methionine and 18.82% aromatic amino acids）. Reverse transcription- polymerase chain reaction analysis indicated that the transcriptional level of PraAry mRNA in P. rapae pupae fat body is inducible in response to parasitism by P. puparum.

关键词： cloning gene expression parasitism Pieris rapae Pteromalus puparum storage protein

Cellular and humoral immune interactions between Drosophila and its parasitoids

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Insect Science 2021年第5期28卷 1208-1227页

作者： Lei Yang Li-Ming Qiu Qi Fang David W.Stanley gong-yin ye State Key Laboratory of Rice Biology&Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects Institute of Insect SciencesZhejiang University.HangzhouChina USD A Agricultural Research Service Biological Control of Insects Research LaboratoryColumbiaMissouriUnited States

The immune interactions occurring between parasitoids and their host insects,especially in Drosophila-wasp models,have long been the research focus of insect immunology and *** infestation in Drosophila is counteracted by its multiple natural immune defense systems,which include cellular and humoral immunity,Occurring in the hemocoel,cellular immune responses involve the proliferation,differentiation,migration and spreading of host hemocytes and parasitoid encapsulation by ***,humoral immune responses rely more heavily on melanization and on the Toll,Imd and Jak/Stat immune pathways associated w ith antimicrobial peptides along with stress *** the wasps'side,successful development is achieved by introducing various virulence factors to counteract immune responses of *** or all of these factors manipulate the host's immunity for successful *** we review current knowledge of the cellular and humoral immune interactions between Drosophila and its parasitoids,focusing on the defense mechanisms used by Drosophila and the strategies evolved by parasitic wasps to outwit it.

关键词： Drosophilas immunity parasitoid venom virus-like particles

Accumulation of cadmium and its effects on growth, development and hemolymph biochemical compositions in Boettcherisca peregrina larvae （Diptera： Sarcophagidae）

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Insect Science 2006年第1期13卷 31-39页

作者： GUO-XING WU gong-yin ye CUI HU JIA-AN CHENG Institute of Applied Entomology College of Agriculture and Biotechnology Zhejiang University Hangzhou China

Newly emerged larvae of the fleshfly, Boettcherisca peregrina were exposed to two different CdCl2 concentrations of 100μg/g and 400 mg/g diet fresh weight （DFW）. They were administered in the diets until the end of larval stage. Cd-exposed larvae accumulated significant amounts of Cd and this accumulation increased with the exposure dose and time. The body weights were lightened and lengths of larvae were shortened considerably after Cd exposure, especially at the higher Cd concentration. The total larval duration was also extremely affected due to Cd exposure. The average duration was prolonged significantly by 14 h at the lower Cd concentration, while it was increased by 33.7 h over controls at the higher Cd concentration. A significant decrease in contents of either soluble proteins, total lipids or caloric values in the hemolymph occurred due to Cd exposure throughout the entire tested period but after 120 h of Cd exposure. In contrast, when exposed to Cd with its higher concentration, total sugar contents in the hemolymph were increased strikingly over the whole tested time, except after 96 h of Cd exposure, while they were not apparently altered except after 24 h of Cd exposure at the lower concentration. Thus, it is suggested that Cd exposure shows significant adverse impact on the growth and development, as well as metabolism, in larvae of this fleshfly, depending on its exposure time and dose.

关键词： Boettcherisca peregrina cadmium caloric value growth and development,lipids proteins total sugar