Functional analysis of a NF-κB transcription factor in the immune defense of Oriental fruit fly, Bactrocera dorsalis Hendel (Diptera: Tephritidae)

AbstractAlthough some novel antimicrobial peptides (AMP) have been successfully isolated from Bactrocera dorsalis Hendel, the mechanisms underlying the induction of these peptides are still elusive. The homolog of NF-κB transcription factor Relish, designated as BdRelish, was cloned from B. dorsalis. The full length cDNA of BdRelish is 3954 bp with an open reading frame that encodes 1013 amino acids. Similar to Drosophila Relish and the mammalian p100, it is a compound protein containing a conserved Rel homology domain, an IPT (Ig-like, plexins, transcription factors) domain and an IκB-like domain (four ankyrin repeats), the nuclear localization signal RKRRR is also detected at the residues 449–453, suggesting that it has homology to Relish and it is a member of the Rel family of transcription activator proteins. Reverse transcription quantitative polymerase chain reaction analysis reveals that BdRelish mRNAs are detected in different quantities from various tissues and the highest transcription level of BdRelish is determined in fat body. The injection challenge of Escherichia coli and Staphylococcus aureas significantly upregulated the expression of BdRelish. The injection of BdRelish dsRNA markedly reduced the expression of BdRelish and decreased the transcription magnitude of antimicrobial peptides. Individuals injected BdRelish dsRNA died at a significantly faster rate compared with the control groups. Therefore, BdRelish is vital for the transcription of AMPs to attack the invading bacteria.

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Molecular cloning, expression, and characterization of E2F transcription factor 4 from Antheraea pernyi

Bulletin of Entomological Research ◽

10.1017/s0007485317000426 ◽

2017 ◽

Vol 107 (6) ◽

pp. 839-846 ◽

Cited By ~ 2

Author(s):

M.N. Abbas ◽

S. Kausar ◽

Y.-X. Sun ◽

Y. Sun ◽

L. Wang ◽

...

Keyword(s):

Escherichia Coli ◽

Transcription Factor ◽

Amino Acid ◽

Amino Acid Sequence ◽

Fat Body ◽

Micrococcus Luteus ◽

Polymerase Chain Reaction Analysis ◽

Antheraea Pernyi ◽

E2f Transcription Factor ◽

Transcription Factor 4

AbstractThe E2F transcription factor family is distributed widely in eukaryotes and has been well studied among mammals. In the present study, the E2F transcription factor 4 (E2F4) gene was isolated from fat bodies of Antheraea pernyi and sequenced. E2F4 comprised a 795 bp open reading frame encoding a deduced amino acid sequence of 264 amino acid residues. The recombinant protein was expressed in Escherichia coli (Transetta DE3), and anti-E2F4 antibodies were prepared. The deduced amino acid sequence displayed significant homology to an E2F4-like protein from Bombyx mori L. Quantitative real-time polymerase chain reaction analysis revealed that E2F4 expression was highest in the integument, followed by the fat body, silk glands, and haemocytes. The expression of E2F4 was upregulated in larvae challenged by bacterial (Escherichia coli, Micrococcus luteus), viral (nuclear polyhedrosis virus), and fungal (Beauveria bassiana) pathogens. These observations indicated that E2F4 is an inducible protein in the immune response of A. pernyi and probably in other insects.

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Molecular Characteristics of Fat Body Protein 1 in the Oriental Fruit Fly, Bactrocera dorsalis

Insects ◽

10.3390/insects12040319 ◽

2021 ◽

Vol 12 (4) ◽

pp. 319

Author(s):

Yao-Chih Yu ◽

Hsuan Lu ◽

Yi-Cheng Chiang ◽

Cheng-Lung Tsai ◽

Yu-Han Zuo ◽

...

Keyword(s):

Fat Body ◽

Expression Profiles ◽

Fruit Fly ◽

Genome Project ◽

Bactrocera Dorsalis ◽

Molecular Characteristics ◽

Body Protein ◽

Larval Stages ◽

Rnai Treatment ◽

Third Instar Larvae

Bactrocera dorsails fat body protein 1 (Bdfbp1) cDNA was cloned (GenBank accession no. MT514270), and the complete 3,749-bp cDNA encoded a 1,152-amino acid protein. The phylogenetic relationship of dipteran fbp1s was analyzed. The sequence XP_028900815 from the insect genome project for Zeugodacus cucurbitae (LOC105219342) was proposed that two fbp1 genes were present in the sequence. The developmental transcriptional expression profiles were determined. In the larval stages, Bdfbp1 mRNA had significantly higher expression in the late third instar larvae compared with first, second, and early third instar larvae. In the pupal stages, the highest expression of Bdfbp1 mRNA was found in the newly pupated pupae and then decreased with age. In the fat body of female adults, Bdfbp1 was highly expressed in newly emerged samples and decreased rapidly over the following three days. In the fat body of male adults, Bdfbp1 was highly expressed in newly eclosed samples. RNAi treatment decreased the expression level of Bdfbp1 without statistical difference. However, RNAi treatment significantly decreased the rate of eclosion. These results suggest that Bdfbp1 may function as a storage protein and be associated with adult eclosion.

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Proteomic Identification of Immune-Related Silkworm Proteins Involved in the Response to Bacterial Infection

Journal of Insect Science ◽

10.1093/jisesa/iez056 ◽

2019 ◽

Vol 19 (4) ◽

Cited By ~ 1

Author(s):

Shiyi Chen ◽

Zhaoming Dong ◽

Xiu Ren ◽

Dongchao Zhao ◽

Yan Zhang ◽

...

Keyword(s):

Fat Body ◽

Quantitative Polymerase Chain Reaction ◽

Immune Defense ◽

Effector Proteins ◽

Control Group ◽

Transcriptional Level ◽

Signaling Proteins ◽

E Coli ◽

A Genome ◽

Immunological Recognition

Abstract Bombyx mori (Lepidoptera: Bombycidae) is an important economic insect and a classic Lepidopteran model system. Although immune-related genes have been identified at a genome-wide scale in the silkworm, proteins involved in immune defense of the silkworm have not been comprehensively characterized. In this study, two types of bacteria were injected into the silkworm larvae, Gram-negative Escherichia coli (Enterobacteriales: Enterobacteriaceae), or Gram-positive Staphylococcus aureus (Bacillales: Staphylococcaceae). After injection, proteomic analyses of hemolymph were performed by liquid chromatography—tandem mass spectrometry. In total, 514 proteins were identified in the uninduced control group, 540 were identified in the E. coli-induced group, and 537 were identified in the S. aureus-induced group. Based on Uniprot annotations, 32 immunological recognition proteins, 28 immunological signaling proteins, and 21 immunological effector proteins were identified. We found that 127 proteins showed significant upregulation, including 10 immunological recognition proteins, 4 immunological signaling proteins, 11 immunological effector proteins, and 102 other proteins. Using real-time quantitative polymerase chain reaction in the fat body, we verified that immunological recognition proteins, signaling proteins, and effector proteins also showed significant increases at the transcriptional level after infection with E. coli and S. aureus. Five newly identified proteins showed upregulation at both protein and transcription levels after infection, including 30K protein, yellow-d protein, chemosensory protein, and two uncharacterized proteins. This study identified many new immune-related proteins, deepening our understanding of the immune defense system in B. mori. The data have been deposited to the iProX with identifier IPX0001337000.

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DIFFERENTIAL GENE EXPRESSION IN MALE AND FEMALE FAT BODY IN THE ORIENTAL FRUIT FLY,Bactrocera dorsalis

Archives of Insect Biochemistry and Physiology ◽

10.1002/arch.21142 ◽

2013 ◽

Vol 85 (1) ◽

pp. 48-59 ◽

Cited By ~ 6

Author(s):

Yu-Han Zuo ◽

Mei-Er Chen

Keyword(s):

Gene Expression ◽

Differential Gene Expression ◽

Fat Body ◽

Fruit Fly ◽

Bactrocera Dorsalis ◽

Oriental Fruit Fly ◽

Male And Female ◽

Differential Gene

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De novo Cloning and Annotation of Genes Associated with Immunity, Detoxification and Energy Metabolism from the Fat Body of the Oriental Fruit Fly, Bactrocera dorsalis

PLoS ONE ◽

10.1371/journal.pone.0094470 ◽

2014 ◽

Vol 9 (4) ◽

pp. e94470 ◽

Cited By ~ 21

Author(s):

Wen-Jia Yang ◽

Guo-Rui Yuan ◽

Lin Cong ◽

Yi-Fei Xie ◽

Jin-Jun Wang

Keyword(s):

Energy Metabolism ◽

De Novo ◽

Fat Body ◽

Fruit Fly ◽

Bactrocera Dorsalis ◽

Oriental Fruit Fly

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Humoral immunocompetence shifts in response to developmental stage change and mating access in Bactrocera dorsalis Hendel (Diptera: Tephritidae)

Bulletin of Entomological Research ◽

10.1017/s0007485314000911 ◽

2015 ◽

Vol 105 (2) ◽

pp. 166-172 ◽

Cited By ~ 2

Author(s):

Z. Shi ◽

Y. Lin ◽

Y. Hou ◽

H. Zhang

Keyword(s):

Antibacterial Activity ◽

Developmental Stage ◽

Maximum Level ◽

Fruit Fly ◽

Investment Strategy ◽

Immune Defense ◽

Bactrocera Dorsalis ◽

Male And Female ◽

Immune Defenses ◽

Stage Change

AbstractBecause immune defenses are often costly employed, insect immunocompetence cannot be always maintained at its maximum level. Here, the oriental fruit fly, Bactrocera dorsalis (Hendel), was used as a study object to investigate how its immune defenses varied with the developmental stage change and mating access. Our data indicated that both phenoloxidase (PO) activity and antibacterial activity significantly increased from new larvae to pupae but decreased in adults after emergence. Furthermore, both the PO activity and antibacterial activity in the hemolymph of copulated male and female adults were dramatically higher than that of virgin male and female ones, respectively. It provided the evidence that copulation could increase the magnitude of immune defense in hemolymph of B. dorsalis. Together, these results suggest that B. dorsalis possess a flexible investment strategy in immunity to meet its specific needs based on the endo- and exogenous factors, such as their distinct food source and living environments.

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The hAT-family transposable element, hopper, from Bactrocera dorsalis is a functional vector for insect germline transformation

BMC Genetics ◽

10.1186/s12863-020-00942-3 ◽

2020 ◽

Vol 21 (S2) ◽

Author(s):

Alfred M. Handler ◽

Marc F. Schetelig

Keyword(s):

Transposable Element ◽

Fluorescent Protein ◽

Fruit Fly ◽

Bactrocera Dorsalis ◽

Helper Plasmid ◽

Reading Frame ◽

Germline Transformation ◽

Functional Genomic Analysis ◽

Wide Range ◽

Dipteran Species

Abstract Background The hopper hAT-family transposable element isolated from the Oriental fruit fly, Bactrocera dorsalis, is distantly related to both the Drosophila hobo element and the Activator element from maize. The original 3120 bp hopperBd-Kah element isolated from the Kahuku wild-type strain was highly degenerate and appeared to have a mutated transposase and terminal sequences, while a second 3131 bp element, hopperBd-we, isolated from a white eye mutant strain had an intact transposase reading frame and terminal sequences consistent with function. Results The hopperBd-we element was tested for function by its ability to mediate germline transformation in two dipteran species other than B. dorsalis. This was achieved by creating a binary vector/helper transformation system by linking the hopperBd-we transposase reading frame to a D. melanogaster hsp70 promoter for a heat-inducible transposase helper plasmid, and creating vectors marked with the D. melanogaster mini-white+ or polyubiquitin-regulated DsRed fluorescent protein markers. Conclusions Both vectors were successfully used to transform D. melanogaster, and the DsRed vector was also used to transform the Caribbean fruit fly, Anastrepha suspensa, indicating a wide range of hopper function in dipteran species and, potentially, non-dipteran species. This vector provides a new tool for insect genetic modification for both functional genomic analysis and the control of insect populations.

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