Dataset Information

Expression and functional analysis of P450 gene induced tolerance/resistance to lambda-cyhalothrin in quercetin fed larvae of beet armyworm Spodoptera exigua (Hubner).

ABSTRACT: Beet armyworm, Spodoptera exigua (Hübner) is an agronomical important and most devastating polyphagous pest that damages a variety of crops around the globe including China. Quercetin is one of the abundant dietary flavonoids and the important defense allelochemicals in plants. Therefore, the changes in insect detoxification enzymes activities in response to plants allelochemicals may result increased the sensitivity to insecticides. In this study, we examined the induced effect of quercetin on larval tolerance to lambda-cyhalothrin in S. exigua. Application of cytochrome P450 inhibitor piperonyl butoxide (PBO) significantly synergized the lambda-cyhalothrin toxicity in quercetin-fed S. exigua larvae. Moreover, larval weight significantly reduced in quercetin, lambda-cyhalothrin, and quercetin?+?lambda-cyhalothrin treatment. Furthermore, our results showed that the P450 detoxification enzyme effectively increased in all treatments as compared to the control. Quantitative Real-time PCR analysis revealed that expression level of CYP6AE10 significantly upregulated in larvae treated with quercetin, lambda-cyhalothrin and quercetin?+?lambda-cyhalothrin in the midgut and fat body respectively. In addition, RNAi mediated knockdown of CYP6AE10 in S. exigua larvae significantly decreased the transcription level of target cytochrome P450 gene followed by the exposure with quercetin, lambda-cyhalothrin, and quercetin?+?lambda-cyhalothrin. Similarly, the knockdown of CYP6AE10 by the injection of dsRNA led to increased mortality after the treatment with respective chemicals. Overall, these data showed that P450s might possibly play an important role in the metabolic adaptation of S. exigua larvae to its host plant defense allelochemicals as well as insecticides. In conclusion, S. exigua can take benefit from its host plant's secondary metabolites to elaborate its defense against synthetic insecticides.

SUBMITTER: Hafeez M

PROVIDER: S-EPMC6933212 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Expression and functional analysis of P450 gene induced tolerance/resistance to lambda-cyhalothrin in quercetin fed larvae of beet armyworm <i>Spodoptera exigua</i> (Hübner).

Hafeez Muhammad M Qasim Muhammad M Ali Sajjad S Yousaf Hafiz Kamran HK Waqas Muhammad M Ali Ehsan E Ahmad Muhammad Afaq MA Jan Saad S Bashir Muhammad Amjad MA Noman Ali A Wang Mo M Gharmh Hamed A HA Khan Khalid Ali KA

Saudi journal of biological sciences 20190522 1

Beet armyworm, <i>Spodoptera exigua</i> (Hübner) is an agronomical important and most devastating polyphagous pest that damages a variety of crops around the globe including China. Quercetin is one of the abundant dietary flavonoids and the important defense allelochemicals in plants. Therefore, the changes in insect detoxification enzymes activities in response to plants allelochemicals may result increased the sensitivity to insecticides. In this study, we examined the induced effect of querce ...[more]

PMID: 31889821

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Project description:Quantitative real-time PCR (qRT-PCR) is a reliable and reproducible technique for measuring and evaluating changes in gene expression. The most common method for analyzing qRT-PCR data is to normalize mRNA levels of target genes to internal reference genes. Evaluating and selecting stable reference genes on a case-by-case basis is critical. The present study aimed to facilitate gene expression studies by identifying the most suitable reference genes for normalization of mRNA expression in qRT-PCR analysis of the beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae). For this purpose, three software tools (geNorm, NormFinder and BestKeeper) were used to investigate 10 candidate reference genes in nine developmental stages and five different tissues (epidermis, head, midgut, fat body and hemolymph) in three larval physiological stages (molting, feeding and wandering stages) of, S. exigua. With the exception of 18S ribosomal RNA (18S), all other candidate genes evaluated, β-actin1(ACT1), β-actin2 (ACT2), elongation factor1(EF1), elongation factor 2 (EF2), Glyceralde hyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein L10 (L10), ribosomal protein L17A (L17A), superoxide dismutase (SOD), α-tubulin (TUB),proved to be acceptable reference genes. However, their suitability partly differed between physiological stages and different tissues. L10, EF2 and L17A ranked highest in all tissue sample sets. SOD, ACT2, GAPDH, EF1 and ACT1 were stably expressed in all developmental stage sample sets; ACT2, ACT1 and L10 for larvae sample sets; GAPDH, ACT1 and ACT2 for pupae and adults; SOD and L17A for males; and EF2 and SOD for females. The expression stability of genes varied in different conditions. The findings provided here demonstrated, with a few exceptions, the suitability of most of the 10 reference genes tested in tissues and life developmental stages. Overall, this study emphasizes the importance of validating reference genes for qRT-PCR analysis in S. exigua.

Project description:BACKGROUND:To understand the genetic mechanisms of insect herbivory, the transcriptional response of insects feeding on different host plant species has to be studied. Here, we generated gene expression data of the generalist herbivore Spodoptera exigua (Hübner) feeding on three selected host plant species and a control (artificial diet). The host plant species used in this study -cabbage (Brassica oleracea), maize (Zea mays) and tobacco (Nicotiana tabacum)- are members of different plant families that each employ specific defence mechanisms and toxins. RESULTS:Spodoptera exigua larvae had a higher growth rate, indicator for herbivore success, when feeding on Z. mays compared to larvae feeding on B. oleracea or N. tabacum. Larvae feeding on the different host plant species showed divergent transcriptional responses. We identified shared and unique gene expression patterns dependent of the host plant species the larvae fed on. Unique gene expression patterns, containing uniquely upregulated transcripts including specific detoxification genes, were found for larvae feeding on either B. oleracea or N. tabacum. No diet-specific gene cluster was identified for larvae feeding on the host for which larvae showed optimal herbivore success, Z. mays, or artificial diet. In contrast, for larvae feeding on hosts for which they showed low herbivore success, specific diet-dependent gene clusters were identified. Functional annotation of these clusters indicates that S. exigua larvae deploy particular host plant-specific genes for digestion and detoxification. CONCLUSIONS:The lack of a host plant-specific gene activity for larvae feeding on Z. mays and the artificial diet suggest a general and non-specific gene activity for host plants with optimal herbivore success. Whereas the finding of specific gene clusters containing particular digestion and detoxifying genes expressed in larvae feeding on B. oleracea and N. tabacum, with low herbivore success, imply a host plant-specific gene activity for larvae feeding on host plants with suboptimal herbivore success. This observation leads to the conclusion that a polyphagous herbivore is able to feed on a large variation of host plants due to the flexibility and diversity of genes involved in digestion and detoxification that are deployed in response to particular host plant species.

Dataset Information

Expression and functional analysis of P450 gene induced tolerance/resistance to lambda-cyhalothrin in quercetin fed larvae of beet armyworm Spodoptera exigua (Hubner).

Publications

Expression and functional analysis of P450 gene induced tolerance/resistance to lambda-cyhalothrin in quercetin fed larvae of beet armyworm <i>Spodoptera exigua</i> (Hübner).

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