Life Sciences Commons^™

Selection Of Housekeeping Genes And Demonstration Of Rnai In Cotton Leafhopper, Amrasca Biguttula Biguttula (Ishida), Satnam Singh, Mridula Gupta, Suneet Pandher, Gurmeet Kaur, Pankaj Rathore, Subba Reddy Palli

Entomology Faculty Publications

Amrasca biguttula biguttula (Ishida) commonly known as cotton leafhopper is a severe pest of cotton and okra. Not much is known on this insect at molecular level due to lack of genomic and transcriptomic data. To prepare for functional genomic studies in this insect, we evaluated 15 common housekeeping genes (Tub, B-Tub, EF alpha, GADPH, UbiCF, RP13, Ubiq, G3PD, VATPase, Actin, 18s, 28s, TATA, ETF, SOD and Cytolytic actin) during different developmental stages and under starvation stress. We selected early (1^st and 2 …

Temporal Transcriptomic Profiling Of The Ant-Feeding Assassin Bug Acanthaspis Cincticrus Reveals A Biased Expression Of Genes Associated With Predation In Nymphs, Fei Kou, Hu Li, Shujuan Li, Huaizhu Xun, Yinqiao Zhang, Ziqiang Sun, Xuguo Zhou, Wanzhi Cai

Entomology Faculty Publications

Acanthaspis cincticrus (Stål) is an assassin bug with a specialized camouflaging behavior to ambush ants in the nymphal stages. In this study, we comprehensively sequenced all the life stages of A. cincticrus, including the eggs, five nymph instars, female and male adults using Illumina HiSeq technology. We obtained 176 million clean sequence reads. The assembled 84,055 unigenes were annotated and classified functionally based on protein databases. Among the unigenes, 29.03% were annotated by one or more databases, suggesting their well-conserved functions. Comparison of the gene expression profiles in the egg, nymph and adult stages revealed certain bias. Functional enrichment …

20-Hydroxyecdysone (20e) Primary Response Gene E93 Modulates 20e Signaling To Promote Bombyx Larval-Pupal Metamorphosis, Xi Liu, Fangyin Dai, Enen Guo, Kang Li, Li Ma, Ling Tian, Yang Cao, Guozheng Zhang, Subba Reddy Palli, Sheng Li

Entomology Faculty Publications

As revealed in a previous microarray study to identify genes regulated by 20-hydroxyecdysone (20E) and juvenile hormone (JH) in the silkworm, Bombyx mori, E93 expression in the fat body was markedly low prior to the wandering stage but abundant during larval-pupal metamorphosis. Induced by 20E and suppressed by JH, E93 expression follows this developmental profile in multiple silkworm alleles. The reduction of E93 expression by RNAi disrupted 20E signaling and the 20E-induced autophagy, caspase activity, and cell dissociation in the fat body. Reducing E93 expression also decreased the expression of the 20E-induced pupal-specific cuticle protein genes and prevented growth …

Stable Reference Gene Selection For Rt-Qpcr Analysis In Nonviruliferous And Viruliferous Frankliniella Occidentalis, Chunxiao Yang, Hui Li, Huipeng Pan, Yabin Ma, Deyong Zhang, Yong Liu, Zhanhong Zhang, Changying Zheng, Dong Chu

Entomology Faculty Publications

Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is a reliable technique for measuring and evaluating gene expression during variable biological processes. To facilitate gene expression studies, normalization of genes of interest relative to stable reference genes is crucial. The western flower thrips Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), the main vector of tomato spotted wilt virus (TSWV), is a destructive invasive species. In this study, the expression profiles of 11 candidate reference genes from nonviruliferous and viruliferous F. occidentalis were investigated. Five distinct algorithms, geNorm, NormFinder, BestKeeper, the ΔCt method, and RefFinder, were used to determine the performance …

Selection Of Reference Genes For Rt-Qpcr Analysis In The Monarch Butterfly, Danaus Plexippus (L.), A Migrating Bio-Indicator, Huipeng Pan, Xiaowei Yang, Keith Bidne, Richard L. Hellmich, Blair D. Siegfried, Xuguo Zhou

Entomology Faculty Publications

Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is a powerful technique to quantify gene expression. To facilitate gene expression study and obtain accurate results, normalization relative to stably expressed reference genes is crucial. The monarch butterfly, Danaus plexippus (L.), is one of the most recognized insect species for its spectacular annual migration across North America. Besides its great voyages, D. plexippus has drawn attention to its role as a bio-indicator, ranging from genetically modified organisms (GMOs) to natural ecosystems. In this study, nine reference genes from D. plexippus genome were selected as the candidate reference genes. The expression profiles of these …

Stably Expressed Housekeeping Genes Across Developmental Stages In The Two-Spotted Spider Mite, Tetranychus Urticae, Chunxiao Yang, Huipeng Pan, Yong Liu, Xuguo Zhou

Entomology Faculty Publications

Quantitative real-time PCR (qRT-PCR) is a reliable and reproducible technique for measuring mRNA expression. To facilitate gene expression studies and obtain more accurate qRT-PCR analysis, normalization relative to stable housekeeping genes is mandatory. In this study, ten housekeeping genes, including beta-actin (Actin), elongation factor 1 α (EF1A), glyceralde hyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein L13 (RPL13), ribosomal protein 49 (RP49), α-tubulin (Tubulin), vacuolar-type H+-ATPase (v-ATPase), succinate dehydrogenase subunit A (SDHA) , 28S ribosomal RNA (28S), and 18S ribosomal RNA (18S) from …

Selection Of Reference Genes For Expression Analysis Using Quantitative Real-Time Pcr In The Pea Aphid, Acyrthosiphon Pisum (Harris) (Hemiptera, Aphidiae), Chunxiao Yang, Huipeng Pan, Yong Liu, Xuguo Zhou

Entomology Faculty Publications

To facilitate gene expression study and obtain accurate qRT-PCR analysis, normalization relative to stable expressed housekeeping genes is required. In this study, expression profiles of 11 candidate reference genes, including actin (Actin), elongation factor 1 α (EF1A), TATA-box-binding protein (TATA), ribosomal protein L12 (RPL12), β-tubulin (Tubulin), NADH dehydrogenase (NADH), vacuolar-type H+-ATPase (v-ATPase), succinate dehydrogenase B (SDHB), 28S ribosomal RNA (28S), 16S ribosomal RNA (16S), and 18S ribosomal RNA (18S) from the pea aphid Acyrthosiphon pisum, …

Validation Of Reference Housekeeping Genes For Gene Expression Studies In Western Corn Rootworm (Diabrotica Virgifera Virgifera), Thaís Barros Rodrigues, Chitvan Khajuria, Haichuan Wang, Natalie Matz, Danielle Cunha Cardoso, Fernando Hercos Valicente, Xuguo Zhou, Blair Siegfried

Entomology Faculty Publications

Quantitative Real-time PCR (qRT-PCR) is a powerful technique to investigate comparative gene expression. In general, normalization of results using a highly stable housekeeping gene (HKG) as an internal control is recommended and necessary. However, there are several reports suggesting that regulation of some HKGs is affected by different conditions. The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a serious pest of corn in the United States and Europe. The expression profile of target genes related to insecticide exposure, resistance, and RNA interference has become an important experimental technique for study of western corn rootworms; however, lack …

Identification And Developmental Profiling Of Micrornas In Diamondback Moth, Plutellaxylostella (L.), Pei Liang, Bing Feng, Xuguo Zhou, Xiwu Gao

Entomology Faculty Publications

MicroRNAs (miRNAs) are a group of small RNAs involved in various biological processes through negative regulation of mRNAs at the post-transcriptional level. Although miRNA profiles have been documented in over two dozen insect species, few are agricultural pests. In this study, both conserved and novel miRNAs in the diamondback moth, Plutella xylostella L., a devastating insect pest of cruciferous crops worldwide, were documented. High-throughput sequencing of a small RNA library constructed from a mixed life stages of P. xylostella, including eggs, 1st to 4th (last) instar larvae, pupae and adults, identified 384 miRNAs, of which 174 were P. xylostella specific. …

Exploring Valid Reference Genes For Quantitative Real-Time Pcr Analysis In Plutella Xylostella (Lepidoptera: Plutellidae), Wei Fu, Wen Xie, Zhuo Zhang, Shaoli Wang, Qingjun Wu, Yong Liu, Xiaomao Zhou, Xuguo Zhou, Youjun Zhang

Entomology Faculty Publications

Quantitative real-time PCR (qRT-PCR), a primary tool in gene expression analysis, requires an appropriate normalization strategy to control for variation among samples. The best option is to compare the mRNA level of a target gene with that of reference gene(s) whose expression level is stable across various experimental conditions. In this study, expression profiles of eight candidate reference genes from the diamondback moth, Plutella xylostella, were evaluated under diverse experimental conditions. RefFinder, a web-based analysis tool, integrates four major computational programs including geNorm, Normfinder, BestKeeper, and the comparative ΔCt method to comprehensively rank the tested candidate …

Wolbachia Association With The Tsetse Fly, Glossina Fuscipes Fuscipes, Reveals High Levels Of Genetic Diversity And Complex Evolutionary Dynamics, Rebecca E. Symula, Uzma Alam, Corey L. Brelsfoard, Yineng Wu, Richard Echodu, Loyce M. Okedi, Serap Aksoy, Adalgisa Caccone

Entomology Faculty Publications

BACKGROUND: Wolbachia pipientis, a diverse group of α-proteobacteria, can alter arthropod host reproduction and confer a reproductive advantage to Wolbachia-infected females (cytoplasmic incompatibility (CI)). This advantage can alter host population genetics because Wolbachia-infected females produce more offspring with their own mitochondrial DNA (mtDNA) haplotypes than uninfected females. Thus, these host haplotypes become common or fixed (selective sweep). Although simulations suggest that for a CI-mediated sweep to occur, there must be a transient phase with repeated initial infections of multiple individual hosts by different Wolbachia strains, this has not been observed empirically. Wolbachia has been found in the tsetse fly, Glossina …

Comparative Mitogenomic Analysis Of Damsel Bugs Representing Three Tribes In The Family Nabidae (Insecta: Hemiptera), Hu Li, Haiyu Liu, Fan Song, Aimin Shi, Xuguo Zhou, Wanzhi Cai

Entomology Faculty Publications

BACKGROUND: Nabidae, a family of predatory heteropterans, includes two subfamilies and five tribes. We previously reported the complete mitogenome of Alloeorhynchus bakeri, a representative of the tribe Prostemmatini in the subfamily Prostemmatinae. To gain a better understanding of architecture and evolution of mitogenome in Nabidae, mitogenomes of five species representing two tribes (Gorpini and Nabini) in the subfamily Nabinae were sequenced, and a comparative mitogenomic analysis of three nabid tribes in two subfamilies was carried out.

METHODOLOGY/PRINCIPAL FINDINGS: Nabid mitogenomes share a similar nucleotide composition and base bias, except for the control region, where differences are observed at the …

Widespread And Persistent Invasions Of Terrestrial Habitats Coincident With Larval Feeding Behavior Transitions During Snail-Killing Fly Evolution (Diptera: Sciomyzidae), Eric G. Chapman, Andrey A. Przhiboro, James D. Harwood, Benjamin A. Foote, Walter R. Hoeh

Entomology Faculty Publications

BACKGROUND: Transitions in habitats and feeding behaviors were fundamental to the diversification of life on Earth. There is ongoing debate regarding the typical directionality of transitions between aquatic and terrestrial habitats and the mechanisms responsible for the preponderance of terrestrial to aquatic transitions. Snail-killing flies (Diptera: Sciomyzidae) represent an excellent model system to study such transitions because their larvae display a range of feeding behaviors, being predators, parasitoids or saprophages of a variety of mollusks in freshwater, shoreline and dry terrestrial habitats. The remarkable genus Tetanocera (Tetanocerini) occupies five larval feeding groups and all of the habitat types mentioned above. …

Gene Expression Profiling In The Thiamethoxam Resistant And Susceptible B-Biotype Sweetpotato Whitefly, Bemisia Tabaci, Wen Xie, Xin Yang, Shao-Ii Wang, Qing-Jun Wu, Ni-Na Yang, Ru-Mei Li, Xiao-Guo Jiao, Hui-Peng Pan, Bai-Ming Liu, Yun-Tao Feng, Bao-Yun Xu, Xuguo Zhou, You-Jun Zhang

Entomology Faculty Publications

Thiamethoxam has been used as a major insecticide to control the B-biotype sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). Due to its excessive use, a high level of resistance to thiamethoxam has developed worldwide over the past several years. To better understand the molecular mechanisms underlying this resistance in B. tabaci, gene profiles between the thiamethoxam-resistant and thiamethoxam-susceptible strains were investigated using the suppression subtractive hybridization (SSH) library approach. A total of 72 and 52 upand down-regulated genes were obtained from the forward and reverse SSH libraries, respectively. These expressed sequence tags (ESTs) belong to several functional categories based …

Socio-Environmental And Endocrine Influences On Developmental And Caste-Regulatory Gene Expression In The Eusocial Termite Reticulitermes Flavipes, Matthew R. Tarver, Xuguo Zhou, Michael E. Scharf

Entomology Faculty Publications

BACKGROUND: Strict regulation of caste differentiation, at the molecular level, is thought to be important to maintain social structure in insect societies. Previously, a number of extrinsic and intrinsic factors have been shown to influence caste composition in termite colonies. One important factor is the influence of nestmates; in particular, soldier termites are known to inhibit hormone-dependent worker-to-soldier differentiation. However, soldier influences on nestmates at the molecular level are virtually unknown. Here, to test the hypothesis that soldiers can influence nestmate gene expression, we investigated the impact of four treatments on whole-body gene expression in totipotent Reticulitermes flavipes workers: (i) …

Molecular Evidence For A Functional Ecdysone Signaling System In Brugia Malayi, George Tzertzinis, Ana L. Egaña, Subba Reddy Palli, Marc Robinson-Rechavi, Chris R. Gissendanner, Canhui Liu, Thomas R. Unnasch, Claude V. Maina

Entomology Faculty Publications

BACKGROUND: Filarial nematodes, including Brugia malayi, the causative agent of lymphatic filariasis, undergo molting in both arthropod and mammalian hosts to complete their life cycles. An understanding of how these parasites cross developmental checkpoints may reveal potential targets for intervention. Pharmacological evidence suggests that ecdysteroids play a role in parasitic nematode molting and fertility although their specific function remains unknown. In insects, ecdysone triggers molting through the activation of the ecdysone receptor: a heterodimer of EcR (ecdysone receptor) and USP (Ultraspiracle).

METHODS AND FINDINGS: We report the cloning and characterization of a B. malayi EcR homologue (Bma-EcR). Bma-EcR dimerizes with …