Cloning and characterisation of NADPH-dependent cytochrome P450 reductase gene in the cotton bollworm,<i>Helicoverpa armigera</i>

Zhao, Chunqing; Tang, Tao; Feng, Xuebin; Qiu, Lihong

doi:10.1002/ps.3538

Cited by 26 publications

(21 citation statements)

References 57 publications

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“…The conserved polyadenylation signal (AATAAA) nucleotide sequence of SeCPR is consistent with most other CPRs in eukaryotes ( Guo and Sherman 1996 ; Zhao et al. 2014 ).…”

Section: Discussionsupporting

confidence: 73%

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Isolation and Expression Analysis ofCYP9A11and Cytochrome P450 Reductase Gene in the Beet Armyworm (Lepidoptera: Noctuidae)

Zhao¹,

Feng²,

Tang³

et al. 2015

J Insect Sci

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Cytochrome P450 monooxygenases (CYPs), as an enzyme superfamily, is widely distributed in organisms and plays a vital function in the metabolism of exogenous and endogenous compounds by interacting with its obligatory redox partner, CYP reductase (CPR). A novel CYP gene (CYP9A11) and CPR gene from the agricultural pest insect Spodoptera exigua were cloned and characterized. The complete cDNA sequences of SeCYP9A11 and SeCPR are 1,931 and 3,919 bp in length, respectively, and contain open reading frames of 1,593 and 2,070 nucleotides, respectively. Analysis of the putative protein sequences indicated that SeCYP9A11 contains a heme-binding domain and the unique characteristic sequence (SRFALCE) of the CYP9 family, in addition to a signal peptide and transmembrane segment at the N-terminal. Alignment analysis revealed that SeCYP9A11 shares the highest sequence similarity with CYP9A13 from Mamestra brassicae, which is 66.54%. The putative protein sequence of SeCPR has all of the classical CPR features, such as an N-terminal membrane anchor; three conserved domain flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN), and nicotinamide adenine dinucleotide phosphate (NADPH) domain; and characteristic binding motifs. Phylogenetic analysis revealed that SeCPR shares the highest identity with HaCPR, which is 95.21%. The SeCYP9A11 and SeCPR genes were detected in the midgut, fat body, and cuticle tissues, and throughout all of the developmental stages of S. exigua. The mRNA levels of SeCYP9A11 and SeCPR decreased remarkably after exposure to plant secondary metabolites quercetin and tannin. The results regarding SeCYP9A11 and SeCPR genes in the current study provide foundation for the further study of S. exigua P450 system.

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“…The conserved polyadenylation signal (AATAAA) nucleotide sequence of SeCPR is consistent with most other CPRs in eukaryotes ( Guo and Sherman 1996 ; Zhao et al. 2014 ).…”

Section: Discussionsupporting

confidence: 73%

“…Similar to the conserved motif in other organisms ( Ingelman et al. 1997 ; Zhao et al. 2014 ), the FAD-binding motif of Se CPR consists of three amino acids, Arg 467, Tyr 469, and Ser 470 as well.…”

Section: Resultsmentioning

confidence: 87%

Isolation and Expression Analysis ofCYP9A11and Cytochrome P450 Reductase Gene in the Beet Armyworm (Lepidoptera: Noctuidae)

Zhao¹,

Feng²,

Tang³

et al. 2015

J Insect Sci

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“…Several studies have shown that CPRs were inducible by drugs in other organisms. For instance, the level of CPR mRNA in Helicoverpa armigera was up-regulated in the midgut of 6th instar larvae after treated with phenobarbital (PB) in the diet 42 . Shephard et al 43 reported that the activity of the CPR enzyme in rat liver microsomal membranes increased to 2.7- fold after PB treatment.…”

Section: Discussionmentioning

confidence: 99%

Silencing NADPH-cytochrome P450 reductase results in reduced acaricide resistance in Tetranychus cinnabarinus (Boisduval)

Shi

Zhang

Shen

et al. 2015

Sci Rep

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Cytochrome P450 monooxygenases (P450s) are involved in metabolic resistance to insecticides and require NADPH cytochrome P450 reductase (CPR) to transfer electrons when they catalyze oxidation reactions. The carmine spider mite, Tetranychus cinnabarinus is an important pest mite of crop and vegetable plants worldwide, and its resistance to acaricides has quickly developed. However, the role of CPR on the formation of acaricide-resistance in T. cinnabarinus is still unclear. In this study, a full-length cDNA encoding CPR was cloned and characterized from T. cinnabarinus (designated TcCPR). TcCPR expression was detectable in all developmental stages of T. cinnabarinus, but it’s much lower in eggs. TcCPR was up-regulated and more inducible with fenpropathrin treatment in the fenpropathrin-resistant (FeR) strain compared with the susceptible SS strain. Feeding of double-strand RNA was effective in silencing the transcription of TcCPR in T. cinnabarinus, which resulted in decreasing the activity of P450s and increasing the susceptibility to fenpropathrin in the FeR strain but not in the susceptible strain. The current results provide first evidence that the down-regulation of TcCPR contributed to an increase of the susceptibility to fenpropathrin in resistant mites. TcCPR could be considered as a novel target for the development of new pesticides.

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“…Gossypol-induced P450s exhibited high divergence and at least five of them ( CYP321A1 , CYP9A12 , CYP9A14 , CYP6AE11 and CYP6B7 ) contributed to cotton bollworm ( Helicoverpa armigera ) tolerance of deltamethrin [ 26 ]. Five genes, CYP337B1 , CYP9A12 , CYP9A14 , CYP6AE11 and CYP6B7 showed considerably high expression in the resistant strain of Helicoverpa armigera against fenvalerate pesticide [ 27 , 28 ]. The CYP9A14 and CYP6AE11 were significantly over-expressed in Helicoverpa armigera strains from Burkina Faso and Spain when compared with Heliar, a susceptible strain [ 29 ].…”

Section: Introductionmentioning

confidence: 99%

Salt Stress Effects on Secondary Metabolites of Cotton in Relation to Gene Expression Responsible for Aphid Development

et al. 2015

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Many secondary metabolites have insecticidal efficacy against pests and may be affected by abiotic stress. However, little is known of how plants may respond to such stress as pertains the growth and development of pests. The objective of this study was to determine if and how salt stress on cotton plants affects the population dynamics of aphids. The NaCl treatment (50mM, 100mM, 150mM and 200mM) increased contents of gossypol in cotton by 26.8–51.4%, flavonoids by 22.5–37.6% and tannic by 15.1–24.3% at 7–28 d after salt stress. Compared with non-stressed plants, the population of aphids on 150 and 200 mM NaCl stressed plants was reduced by 46.4 and 65.4% at 7d and by 97.3 and 100% at 14 days after infestation. Reductions in aphid population were possibly attributed to the elevated secondary metabolism under salt stress. A total of 796 clones for aphids transcriptome, 412 clones in the positive- library (TEST) and 384 clones in the reverse-library (Ck), were obtained from subtracted cDNA libraries and sequenced. Gene ontology (GO) functional classification and KEGG pathway analysis showed more genes related to fatty acid and lipid biosynthesis, and fewer genes related to carbohydrate metabolism, amino acid metabolism, energy metabolism and cell motility pathways in TEST than in Ck library, which might be the reason of aphids population reduction. A comparative analysis with qRT-PCR indicated high expression of transcripts CYP6A14, CYP6A13, CYP303A1, NADH dehydrogenase and fatty acid synthase in the TEST group. However, CYP307A1 and two ecdysone-induced protein genes were down regulated. The results indicate that genes of aphids related to growth and development can express at a higher level in reaction to the enhanced secondary metabolism in cotton under salinity stress. The expression of CYP307A1 was positively correlated with the population dynamics of aphids since it was involved in ecdysone synthesis.

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Cloning and characterisation of NADPH-dependent cytochrome P450 reductase gene in the cotton bollworm,Helicoverpa armigera

Abstract: The results indicate that HaCPR is important for the development of H. armigera and may play an essential role in the P450-mediated insecticide resistance of H. armigera to fenvalerate.

Cited by 26 publications

References 57 publications

Isolation and Expression Analysis ofCYP9A11and Cytochrome P450 Reductase Gene in the Beet Armyworm (Lepidoptera: Noctuidae)

Isolation and Expression Analysis ofCYP9A11and Cytochrome P450 Reductase Gene in the Beet Armyworm (Lepidoptera: Noctuidae)

Silencing NADPH-cytochrome P450 reductase results in reduced acaricide resistance in Tetranychus cinnabarinus (Boisduval)

Salt Stress Effects on Secondary Metabolites of Cotton in Relation to Gene Expression Responsible for Aphid Development

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