Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera (Hemiptera: Delphacidae)
Nicotinamide adenine dinucleotide phosphate (NADPH) -cytochrome P450 reductase (CPR) is an electron supplier for various cytochrome P450 monooxygenases. Most P450-mediated catalytic reactions in insects require involvement of CPR, such as detoxification of insecticides and plant secondary metabolite...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2016-07-01
|
| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2015.12.171 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849706446285963264 |
|---|---|
| author | YU Hang LIU Su ZHU Qingzi ZHOU Wenwu LIANG Qingmei SHI Xiaoxiao ZHU Zijie ZHU Zengrong |
| author_facet | YU Hang LIU Su ZHU Qingzi ZHOU Wenwu LIANG Qingmei SHI Xiaoxiao ZHU Zijie ZHU Zengrong |
| author_sort | YU Hang |
| collection | DOAJ |
| description | Nicotinamide adenine dinucleotide phosphate (NADPH) -cytochrome P450 reductase (CPR) is an electron supplier for various cytochrome P450 monooxygenases. Most P450-mediated catalytic reactions in insects require involvement of CPR, such as detoxification of insecticides and plant secondary metabolites. So far, CPR genes have been identified and characterized from many model and non-model insect species. Since insect P450 system is one of the most important metabolic adaptive traits involved in the degradation of xenobiotics and regulation of endogenous substrates. CPR, as the indispensable electron donor of P450 system, has attracted increasing attention as a potential candidate to develop novel chemical inhibitor to manage target insect pests. Rice planthoppers, such as Nilaparvata lugens, Laodelphax striatellus and Sogatella furcifera, are considered to be the most serious pests of rice. Previously, some studies on L. striatellus and N. lugens found that silencing the CPR gene by RNAi technology could increase their sensitivity to insecticides, but little was known about S. furcifera. This work firstly reports the identification of CPR gene in S. furcifera and up-regulation of the CPR transcription by chemical insecticides. The research will facilitate further study on the function of CPR in S. furcifera.In this study, a full-length cDNA encoding CPR was cloned from S. furcifera. The phylogenetic relationships of SfCPR with other insect CPRs were estimated by neighbor-joining method, and its distribution in various tissues and different developmental stages were analyzed by real-time quantitative polymerase chain reaction (qPCR). Finally, after exposure of deltamethrin, buprofezin and imidacloprid at sublethal concentrations for 6,12 and 24 h, the relative expression levels of SfCPR in the third-instar nymphs were investigated.By searching the transcriptome data sets of S. furcifera, a cDNA fragment encoding putative CPR (named SfCPR) was identified. This cDNA fragment was then amplified by PCR and sequenced in order to confirm that the sequence was not chimeric. The SfCPR cDNA contained a complete open reading frame (ORF) of 2 034 bp nucleotides, encoding a protein of 677 amino acid residues. The theoretical isoelectric point (pI) and calculated molecular mass of SfCPR protein are 5.48 and 76.762 ku, respectively. The secondary structure of SfCPR protein showed the marked features of typical CPRs, such as N-terminal transmembrane region, FMN-, FAD- and NADPH-binding domains and conserved catalytic residues. In addition, a transmembrane anchor was predicted in the N-terminus of the protein, indicating that SfCPR is an endoplasmic reticulum-located protein. Phylogenic analysis was used to gain insight into the phylogenetic relationships among CPR proteins from diverse insect species. We found that SfCPR and CPRs from other two planthoppers were clustered together. Real-time quantitative PCR showed that the expression of SfCPR was detectable in all developmental stages and the level in adults was the highest. The SfCPR transcripts were also expressed in all the tested tissues of the adults, and most were strongly expressed in the abdomen. The exposure at sublethal concentrations of deltamethrin, buprofezin and imidacloprid led to significantly elevated expression of SfCPR. The expression of SfCPR was activated soon (6 h) after treatment with buprofezin and imidacloprid, while the response of SfCPR expression to deltamethrin was relatively slow (12 h).In conclusion, this work is the first report about the cDNA sequence information, secondary structure and transcription profiles of CPR gene in the S. furcifera. These findings provide foundation for further research on the physiological function of SfCPR. |
| format | Article |
| id | doaj-art-801d849d5e674ab2800f1206576d958d |
| institution | DOAJ |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2016-07-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-801d849d5e674ab2800f1206576d958d2025-08-20T03:16:11ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552016-07-014239140010.3785/j.issn.1008-9209.2015.12.17110089209Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera (Hemiptera: Delphacidae)YU HangLIU SuZHU QingziZHOU WenwuLIANG QingmeiSHI XiaoxiaoZHU ZijieZHU ZengrongNicotinamide adenine dinucleotide phosphate (NADPH) -cytochrome P450 reductase (CPR) is an electron supplier for various cytochrome P450 monooxygenases. Most P450-mediated catalytic reactions in insects require involvement of CPR, such as detoxification of insecticides and plant secondary metabolites. So far, CPR genes have been identified and characterized from many model and non-model insect species. Since insect P450 system is one of the most important metabolic adaptive traits involved in the degradation of xenobiotics and regulation of endogenous substrates. CPR, as the indispensable electron donor of P450 system, has attracted increasing attention as a potential candidate to develop novel chemical inhibitor to manage target insect pests. Rice planthoppers, such as Nilaparvata lugens, Laodelphax striatellus and Sogatella furcifera, are considered to be the most serious pests of rice. Previously, some studies on L. striatellus and N. lugens found that silencing the CPR gene by RNAi technology could increase their sensitivity to insecticides, but little was known about S. furcifera. This work firstly reports the identification of CPR gene in S. furcifera and up-regulation of the CPR transcription by chemical insecticides. The research will facilitate further study on the function of CPR in S. furcifera.In this study, a full-length cDNA encoding CPR was cloned from S. furcifera. The phylogenetic relationships of SfCPR with other insect CPRs were estimated by neighbor-joining method, and its distribution in various tissues and different developmental stages were analyzed by real-time quantitative polymerase chain reaction (qPCR). Finally, after exposure of deltamethrin, buprofezin and imidacloprid at sublethal concentrations for 6,12 and 24 h, the relative expression levels of SfCPR in the third-instar nymphs were investigated.By searching the transcriptome data sets of S. furcifera, a cDNA fragment encoding putative CPR (named SfCPR) was identified. This cDNA fragment was then amplified by PCR and sequenced in order to confirm that the sequence was not chimeric. The SfCPR cDNA contained a complete open reading frame (ORF) of 2 034 bp nucleotides, encoding a protein of 677 amino acid residues. The theoretical isoelectric point (pI) and calculated molecular mass of SfCPR protein are 5.48 and 76.762 ku, respectively. The secondary structure of SfCPR protein showed the marked features of typical CPRs, such as N-terminal transmembrane region, FMN-, FAD- and NADPH-binding domains and conserved catalytic residues. In addition, a transmembrane anchor was predicted in the N-terminus of the protein, indicating that SfCPR is an endoplasmic reticulum-located protein. Phylogenic analysis was used to gain insight into the phylogenetic relationships among CPR proteins from diverse insect species. We found that SfCPR and CPRs from other two planthoppers were clustered together. Real-time quantitative PCR showed that the expression of SfCPR was detectable in all developmental stages and the level in adults was the highest. The SfCPR transcripts were also expressed in all the tested tissues of the adults, and most were strongly expressed in the abdomen. The exposure at sublethal concentrations of deltamethrin, buprofezin and imidacloprid led to significantly elevated expression of SfCPR. The expression of SfCPR was activated soon (6 h) after treatment with buprofezin and imidacloprid, while the response of SfCPR expression to deltamethrin was relatively slow (12 h).In conclusion, this work is the first report about the cDNA sequence information, secondary structure and transcription profiles of CPR gene in the S. furcifera. These findings provide foundation for further research on the physiological function of SfCPR.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2015.12.171cytochrome P450 reductase<italic>Sogatella furcifera</italic>insecticideexpression profile |
| spellingShingle | YU Hang LIU Su ZHU Qingzi ZHOU Wenwu LIANG Qingmei SHI Xiaoxiao ZHU Zijie ZHU Zengrong Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera (Hemiptera: Delphacidae) 浙江大学学报. 农业与生命科学版 cytochrome P450 reductase <italic>Sogatella furcifera</italic> insecticide expression profile |
| title | Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera (Hemiptera: Delphacidae) |
| title_full | Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera (Hemiptera: Delphacidae) |
| title_fullStr | Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera (Hemiptera: Delphacidae) |
| title_full_unstemmed | Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera (Hemiptera: Delphacidae) |
| title_short | Molecular characterization and expression profiles of cytochrome P450 reductase gene in Sogatella furcifera (Hemiptera: Delphacidae) |
| title_sort | molecular characterization and expression profiles of cytochrome p450 reductase gene in sogatella furcifera hemiptera delphacidae |
| topic | cytochrome P450 reductase <italic>Sogatella furcifera</italic> insecticide expression profile |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2015.12.171 |
| work_keys_str_mv | AT yuhang molecularcharacterizationandexpressionprofilesofcytochromep450reductasegeneinsogatellafurciferahemipteradelphacidae AT liusu molecularcharacterizationandexpressionprofilesofcytochromep450reductasegeneinsogatellafurciferahemipteradelphacidae AT zhuqingzi molecularcharacterizationandexpressionprofilesofcytochromep450reductasegeneinsogatellafurciferahemipteradelphacidae AT zhouwenwu molecularcharacterizationandexpressionprofilesofcytochromep450reductasegeneinsogatellafurciferahemipteradelphacidae AT liangqingmei molecularcharacterizationandexpressionprofilesofcytochromep450reductasegeneinsogatellafurciferahemipteradelphacidae AT shixiaoxiao molecularcharacterizationandexpressionprofilesofcytochromep450reductasegeneinsogatellafurciferahemipteradelphacidae AT zhuzijie molecularcharacterizationandexpressionprofilesofcytochromep450reductasegeneinsogatellafurciferahemipteradelphacidae AT zhuzengrong molecularcharacterizationandexpressionprofilesofcytochromep450reductasegeneinsogatellafurciferahemipteradelphacidae |