Biochemical and molecular characterisation of acetylcholinesterase in four field populations of <i>Bactrocera dorsalis</i> (Hendel) (Diptera: Tephritidae)

Shen, Guangmao; Wang, Xiaona; Dou, Wei; Wang, Jinjun

doi:10.1002/ps.3340

Cited by 29 publications

(22 citation statements)

References 87 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Three alternative splice sites, ASI, ASII and ASIV, were conserved from D . melanogaster [20], [40], but one, ASIII, was reported for the first time. Like the alternative splice variants reported in mammals, the four variants in BdRyR exhibited significant developmental or anatomical regulation.…”

Section: Discussionmentioning

confidence: 99%

“…First strand cDNA was synthesized from 2 µg of each total RNA preparation in a 10 µl reaction mixture using random hexamer primers and an oligo (dT) primer using the PrimeScript RT reagent Kit (TaKaRa). Real-time PCR was conducted in an Mx3000P thermal cycler (Stratagene, La Jolla, CA, USA) with the stable B. dorsalis reference gene α - Tubulin (GU269902) [40]. The quantitative real-time PCR was carried out in 20 µl reaction volumes containing 1 µl cDNA (200 ng/µl), 10 µl iQ™ SYBR Green Supermix (BIO-RAD, Hercules, CA, USA), 1 µl each of forward and reverse primers (0.2 mM), and 7 µl ddH2O.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Molecular Characteristics, mRNA Expression, and Alternative Splicing of a Ryanodine Receptor Gene in the Oriental Fruit Fly, Bactrocera dorsalis (Hendel)

et al. 2014

Self Cite

View full text Add to dashboard Cite

Ryanodine receptors (RyRs) are a distinct class of ligand-gated channels controlling the release of calcium from intracellular stores. The emergence of diamide insecticides, which selectively target insect RyRs, has promoted the study of insect RyRs. In the present study, the full-length RyR cDNA (BdRyR) was cloned and characterized from the oriental fruit fly, Bactrocera dorsalis (Hendel), a serious pest of fruits and vegetables throughout East Asia and the Pacific Rim. The cDNA of BdRyR contains a 15,420-bp open reading frame encoding 5,140 amino acids with a predicted molecular weight of 582.4 kDa and an isoelectric point of 5.38. BdRyR shows a high level of amino acid sequence identity (78 to 97%) to other insect RyR isoforms. All common structural features of the RyRs are present in the BdRyR, including a well-conserved C-terminal domain containing consensus calcium-binding EF-hands and six transmembrane domains, and a large N-terminal domain. Quantitative real-time PCR analyses revealed that BdRyR was expressed at the lowest and highest levels in egg and adult, respectively, and that the BdRyR expression levels in the third instar larva, pupa and adult were 166.99-, 157.56- and 808.56-fold higher, respectively, than that in the egg. Among different adult body parts, the highest expression level was observed in the thorax compared with the head and abdomen. In addition, four alternative splice sites were identified in the BdRyR gene, with the first, ASI, being located in the central part of the predicted second spore lysis A/RyR domain. Diagnostic PCR analyses revealed that alternative splice variants were generated not only in a tissue-specific manner but also in a developmentally regulated manner. These results lay the foundation for further understanding the structural and functional properties of BdRyR, and the molecular mechanisms for target site resistance in B. dorsalis.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Molecular Characteristics, mRNA Expression, and Alternative Splicing of a Ryanodine Receptor Gene in the Oriental Fruit Fly, Bactrocera dorsalis (Hendel)

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…The relative expression levels at various developmental stages were normalized to that of 18S ribosomal RNA 47. α-tubulin was used to normalize the data of tissue expression48 and expression under 20E treatment49.…”

Section: Methodsmentioning

confidence: 99%

Alternative splicing contributes to the coordinated regulation of ferritin subunit levels in Bactrocera dorsalis (Hendel)

Jiang

Cong

Niu

et al. 2014

Sci Rep

Self Cite

View full text Add to dashboard Cite

A constant ratio of ferritin heavy chain homolog (HCH) and light chain homolog (LCH) subunits seems to be required to compose the ferritin heteropolymer protein in insects. However, the mechanism by which insect LCH genes regulate protein levels remains unclear. We report that alternative promoters and alternative splicing contribute to maintaining a constant ratio of the two subunits, BdFer1HCH and BdFer2LCH (ferritin 1 HCH and ferritin 2 LCH), in Bactrocera dorsalis, a notorious quarantine pest. The genes BdFer1HCH and BdFer2LCH were identified with a series of potential transcription factor binding sites and were shown to be clustered within the genome in a “head to head” fashion. Thus, we unearthed a potential post-transcriptional mechanism to regulate the levels of LCH subunits, and confirmed that the expressions of BdFer1HCH and BdFer2LCH were induced by 20-hydroecdysone, iron overload, and immune challenge.

show abstract

“…First strand cDNA was synthesized from 2 mg of each total RNA preparation in a 10 ml reaction mixture using random hexamer primers and an oligo (dT) primer using the PrimeScript RT reagent Kit (TaKaRa). Real-time PCR was conducted in an Mx3000P thermal cycler (Stratagene, La Jolla, CA, USA) with the stable B. dorsalis reference gene a-Tubulin (GU269902) [40]. The quantitative real-time PCR was carried out in 20 ml reaction volumes containing 1 ml cDNA (200 ng/ml), 10 ml iQ TM SYBR Green Supermix (BIO-RAD, Hercules, CA, USA), 1 ml each of forward and reverse primers (0.2 mM), and 7 ml ddH2O.…”

Section: Sequence Analysis and Phylogenetic Tree Constructionmentioning

confidence: 99%

“…In the present study, four alternative splice sites were found in the BdRyR gene. Three alternative splice sites, ASI, ASII and ASIV, were conserved from D. melanogaster [20,40], but one, ASIII, was reported for the first time. Like the alternative splice variants reported in mammals, the four variants in BdRyR exhibited significant developmental or anatomical regulation.…”

mentioning

confidence: 99%

Molecular characteristics, mRNA expression, and alternative splicing of a ryanodine receptor gene in the Oriental fruit fly,Bactrocera dorsalis(Hendel)

Yuan¹

2016

2016 International Congress of Entomology

View full text Add to dashboard Cite

Ryanodine receptors (RyRs) are a distinct class of ligand-gated channels controlling the release of calcium from intracellular stores. The emergence of diamide insecticides, which selectively target insect RyRs, has promoted the study of insect RyRs. In the present study, the full-length RyR cDNA (BdRyR) was cloned and characterized from the oriental fruit fly, Bactrocera dorsalis (Hendel), a serious pest of fruits and vegetables throughout East Asia and the Pacific Rim. The cDNA of BdRyR contains a 15,420-bp open reading frame encoding 5,140 amino acids with a predicted molecular weight of 582.4 kDa and an isoelectric point of 5.38. BdRyR shows a high level of amino acid sequence identity (78 to 97%) to other insect RyR isoforms. All common structural features of the RyRs are present in the BdRyR, including a well-conserved C-terminal domain containing consensus calcium-binding EF-hands and six transmembrane domains, and a large N-terminal domain. Quantitative real-time PCR analyses revealed that BdRyR was expressed at the lowest and highest levels in egg and adult, respectively, and that the BdRyR expression levels in the third instar larva, pupa and adult were 166.99-, 157.56-and 808.56-fold higher, respectively, than that in the egg. Among different adult body parts, the highest expression level was observed in the thorax compared with the head and abdomen. In addition, four alternative splice sites were identified in the BdRyR gene, with the first, ASI, being located in the central part of the predicted second spore lysis A/RyR domain. Diagnostic PCR analyses revealed that alternative splice variants were generated not only in a tissue-specific manner but also in a developmentally regulated manner. These results lay the foundation for further understanding the structural and functional properties of BdRyR, and the molecular mechanisms for target site resistance in B. dorsalis.

show abstract

Biochemical and molecular characterisation of acetylcholinesterase in four field populations of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

Abstract: The results suggest that the decrease in susceptibility of B. dorsalis was mainly caused by decrease in AChE activity, and they provide a broad view on the relation between AChE and resistance.

Cited by 29 publications

References 87 publications

Molecular Characteristics, mRNA Expression, and Alternative Splicing of a Ryanodine Receptor Gene in the Oriental Fruit Fly, Bactrocera dorsalis (Hendel)

Molecular Characteristics, mRNA Expression, and Alternative Splicing of a Ryanodine Receptor Gene in the Oriental Fruit Fly, Bactrocera dorsalis (Hendel)

Alternative splicing contributes to the coordinated regulation of ferritin subunit levels in Bactrocera dorsalis (Hendel)

Molecular characteristics, mRNA expression, and alternative splicing of a ryanodine receptor gene in the Oriental fruit fly,Bactrocera dorsalis(Hendel)

Contact Info

Product

Resources

About