scholarly journals Selection and evaluation of qPCR reference genes for expression analysis in the tiny egg parasitoid wasp, Trichogramma dendrolimi  Matsumura (Hymenoptera: Trichogrammatidae)

2021 ◽  
Author(s):  
Liang-xiao Huo ◽  
Xue-ping Bai ◽  
Wu-Nan Che ◽  
Su-fang Ning ◽  
Lin Lv ◽  
...  
Keyword(s):  
Gene Expression ◽  
Biological Control ◽  
Developmental Stage ◽  
Reference Genes ◽  
Biological Control Agent ◽  
Parasitoid Wasp ◽  
Housekeeping Genes ◽  
Egg Parasitoid ◽  
Specific Reference ◽  
Trichogramma Dendrolimi

The egg parasitoid Trichogramma  spp. is an important biological control agent used against multiple species of Lepidopteran pest in forestry and agriculture. Due to the importance of Trichogramma  spp. in biocontrol programs, its biological characteristics have been studied in detail, and current investigations should focus on the molecular biology of these tiny parasitoids. Real-time quantitative PCR (qPCR) is considered as the standard method for quantifying the gene expression of organisms. Surprisingly, the appropriate reference genes to ensure robust qPCR have not been documented at all for the Trichogramma  genus. This study aimed to identify suitable reference genes for use in qPCR procedure of Trichogramma   dendrolimi . Nine candidate housekeeping genes, namely glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ), forkhead box O ( FOXO ), superoxide dismutase ( SOD ), beta-actin ( ACTIN ), ribosomal protein L10a ( RPL10a ), L18 ( RPL18 ), L28 ( RPL28 ), S13 ( RPS13 ), and S15 ( RPS15 ), were tested for their suitability as reference genes for developmental stage (3 rd , 4 th , 5 th , 6 th , 7 th , 8 th , 9 th , and 10 th  day after parasitization), tissue (head, thorax, and abdomen of adults), sex of adults (male and female), and temperature (17?, 25?, and 32?). According to the GeNorm analysis, robust analysis should involve using an appropriate combination of reference genes, namely, at least three genes for different development stages, two genes for different tissues, two genes for different sex, and two genes for different temperature, respectively. According to the RelFinder method and by assessing the integrated values from using the ?Ct method, GeNorm, NormFinder, and BestKeeper, we identified the developmental stage-specific reference genes SOD , GAPDH , and ACTIN ; tissue-specific reference genes RPL18  and RPS15 ; sex-specific reference genes SOD  and RPL18 ; and temperature-specific reference genes RPL18  and RPL10 . When testing the use of stable vs. unstable reference genes, the substantial differences were observed in the estimation expression of a hypothetical target gene, HSP90 , in response to temperature. The present study provides a robust method for the measurement of gene expression in T. dendrolimi  and will be helpful for future biological control programs using Trichogramma  wasps.

scholarly journals Psix striaticeps (Dodd) (Hymenoptera, Scelionidae): an Old World parasitoid of stink bug eggs arrives in Florida, USA

2021 ◽  
Vol 87 ◽  
pp. 503-521
Author(s):  
Sarah Birkmire ◽  
Cory Penca ◽  
Elijah J. Talamas ◽  
Matthew R. Moore ◽  
Amanda C. Hodges
Keyword(s):  
Biological Control ◽  
Biological Control Agent ◽  
Parasitoid Wasp ◽  
Egg Parasitoid ◽  
Control Agent ◽  
Western Hemisphere ◽  
Stink Bug ◽  
Stink Bugs ◽  
Old World ◽  
Piezodorus Guildinii

Psix striaticeps (Dodd) is an egg-parasitoid wasp previously known only from the Old World. We report this species from twelve counties in Florida, which are the first records in the Western Hemisphere. It was collected in yellow cylinder traps and reared from the eggs of three stink bug species: Nezara viridula L., Chinavia marginata (Palisot de Beauvois), and Piezodorus guildinii (Westwood). A COI barcode analysis found a 100% match between the Floridian population and a specimen from South Africa. The prospects of using Ps. striaticeps as a biological control agent against exotic stink bugs are discussed.

scholarly journals Selection and Validation of Reference Genes for Gene Expression Analysis in Tuta absoluta Meyrick (Lepidoptera: Gelechiidae)

Insects ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 589
Author(s):  
Xin Yan ◽  
Yibo Zhang ◽  
Kangkang Xu ◽  
Yawei Wang ◽  
Wenjia Yang
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Housekeeping Genes ◽  
Tuta Absoluta ◽  
Leaf Miner ◽  
Specific Reference ◽  
Internal Reference ◽  
Experimental Conditions

The tomato leaf miner, Tuta absoluta is a destructive pest of tomato. The leaf-mining activities of its larvae can cause significant yield losses. Real-time quantitative polymerase chain reaction (RT-qPCR) is commonly used to measure gene expression, and the selection of stable reference genes for calibration and standardization is critical for accurate use of RT-qPCR. We studied the stable expression of nine common housekeeping genes in T. absoluta. These were examined at different developmental stages, in larval tissues, as well as those induced by exposure to 20E and insecticides. Four dedicated algorithms (geNorm, BestKeeper, NormFinder, and ΔCt method) and online tool (RefFinder) were used to analyze and rank the tested reference genes. Based on the standardized gene expression data of target gene ecdysone receptor (EcR), the applicability of specific reference genes was verified. The results clarify that the optimal internal reference genes vary greatly under different experimental conditions. GAPDH and RPS11 were the best reference genes for developmental stages; RPL28 and RPL10 for different tissues; EF1α and RPL28 for 20E treatment; EF1α and RPL7A for insecticide treatments. The most suitable reference genes in all experimental conditions are EF1α and RPL28.

scholarly journals Sexual dimorphism and sex-biased gene expression in an egg parasitoid species, Anastatus disparis

2020 ◽  
Author(s):  
P.C. Liu ◽  
DeJun Hao ◽  
Hao-Yuan Hu ◽  
Jian-Rong Wei
Keyword(s):  
Gene Expression ◽  
Biological Control ◽  
Molecular Mechanisms ◽  
Parasitoid Wasp ◽  
Egg Parasitoid ◽  
Future Research ◽  
Sexually Dimorphic ◽  
Large Set ◽  
Pest Species ◽  
Expression Of Genes

Abstract BackgroundDifferences in the expression of genes present in both sexes are assumed to contribute to sex differences including behavioural, physiological and morphological dimorphisms. For enriching our knowledge of gender differences in an important egg parasitoid wasp, Anastatus disparis (Hymenoptera: Eupelmidae), sex-biased differences in gene expression were investigated using Illumina-based transcriptomic analysis. ResultsA total of 15812 resulting unigenes were annotated, and a large set of genes accounting for 50.09% of the total showed sex-biased expression and included 630 sex-specific genes. Gene Ontology (GO) enrichment analyses showed that the functional categories associated with sex-biased genes were mainly related to reproduction. In addition, the transcriptome data provided evidence that sex pheromones in A. disparis are produced by the female, and activity of D12-desaturases appear to have been replaced by D9-desaturases playing roles in sex pheromone production. The large set of sex-biased genes identified in this study provide a molecular background for sexually dimorphic traits such as flyability, longevity, and aggression in this species and suggests candidate venom proteins expressed only in females that could be used for biological control.ConclusionsThis study provides comprehensive insight into sexually dimorphic traits of a parasitoid wasp and can inform future research into the molecular mechanisms underlying such traits and the application of parasitoids to the biological control of pest species.

scholarly journals Sexual dimorphism and sex-biased gene expression in an egg parasitoid species, Anastatus disparis

2020 ◽  
Author(s):  
P.C. Liu ◽  
DeJun Hao ◽  
Hao-Yuan Hu ◽  
Jian-Rong Wei
Keyword(s):  
Gene Expression ◽  
Biological Control ◽  
Molecular Mechanisms ◽  
Parasitoid Wasp ◽  
Egg Parasitoid ◽  
Future Research ◽  
Sexually Dimorphic ◽  
Large Set ◽  
Pest Species ◽  
Expression Of Genes

Abstract Background: Differences in the expression of genes present in both sexes are assumed to contribute to sex differences including behavioural, physiological and morphological dimorphisms. For enriching our knowledge of gender differences in an important egg parasitoid wasp, Anastatus disparis (Hymenoptera: Eupelmidae), sex-biased differences in gene expression were investigated using Illumina-based transcriptomic analysis. Results: A total of 15812 resulting unigenes were annotated, and a large set of genes accounting for 50.09% of the total showed sex-biased expression and included 630 sex-specific genes. Gene Ontology (GO) enrichment analyses showed that the functional categories associated with sex-biased genes were mainly related to reproduction. In addition, the transcriptome data provided evidence that sex pheromones in A. disparis are produced by the female, and activity of D12-desaturases appear to have been replaced by D9-desaturases playing roles in sex pheromone production. The large set of sex-biased genes identified in this study provide a molecular background for sexually dimorphic traits such as flyability, longevity, and aggression in this species and suggests candidate venom proteins expressed only in females that could be used for biological control.Conclusions: This study provides comprehensive insight into sexually dimorphic traits of a parasitoid wasp and can inform future research into the molecular mechanisms underlying such traits and the application of parasitoids to the biological control of pest species.

scholarly journals Spatio-temporal selection of reference genes in the two congeneric species of Glycyrrhiza

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yuping Li ◽  
Xiaoju Liang ◽  
Xuguo Zhou ◽  
Yu An ◽  
Ming Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Developmental Stage ◽  
Reference Genes ◽  
Developmental Stages ◽  
Individual Factors ◽  
Congeneric Species ◽  
Spatio Temporal ◽  
The Stability ◽  
Different Tissues ◽  
Selection Of

AbstractGlycyrrhiza, a genus of perennial medicinal herbs, has been traditionally used to treat human diseases, including respiratory disorders. Functional analysis of genes involved in the synthesis, accumulation, and degradation of bioactive compounds in these medicinal plants requires accurate measurement of their expression profiles. Reverse transcription quantitative real-time PCR (RT-qPCR) is a primary tool, which requires stably expressed reference genes to serve as the internal references to normalize the target gene expression. In this study, the stability of 14 candidate reference genes from the two congeneric species G. uralensis and G. inflata, including ACT, CAC, CYP, DNAJ, DREB, EF1, RAN, TIF1, TUB, UBC2, ABCC2, COPS3, CS, R3HDM2, were evaluated across different tissues and throughout various developmental stages. More importantly, we investigated the impact of interactions between tissue and developmental stage on the performance of candidate reference genes. Four algorithms, including geNorm, NormFinder, BestKeeper, and Delta Ct, were used to analyze the expression stability and RefFinder, a comprehensive software, provided the final recommendation. Based on previous research and our preliminary data, we hypothesized that internal references for spatio-temporal gene expression are different from the reference genes suited for individual factors. In G. uralensis, the top three most stable reference genes across different tissues were R3HDM2, CAC and TUB, while CAC, CYP and ABCC2 were most suited for different developmental stages. CAC is the only candidate recommended for both biotic factors, which is reflected in the stability ranking for the spatio (tissue)-temporal (developmental stage) interactions (CAC, R3HDM2 and DNAJ). Similarly, in G. inflata, COPS3, R3HDM2 and DREB were selected for tissues, while RAN, COPS3 and CS were recommended for developmental stages. For the tissue-developmental stage interactions, COPS3, DREB and ABCC2 were the most suited reference genes. In both species, only one of the top three candidates was shared between the individual factors and their interactions, specifically, CAC in G. uralensis and COPS3 in G. inflata, which supports our overarching hypothesis. In summary, spatio-temporal selection of reference genes not only lays the foundation for functional genomics research in Glycyrrhiza, but also facilitates these traditional medicinal herbs to reach/maximize their pharmaceutical potential.

scholarly journals The Effect of Aspergillus Thermomutatus Chrysovirus 1 on the Biology of Three Aspergillus Species

Viruses ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 539 ◽  
Author(s):  
Mahjoub A. Ejmal ◽  
David J. Holland ◽  
Robin M. MacDiarmid ◽  
Michael N. Pearson
Keyword(s):  
Gene Expression ◽  
Biological Control ◽  
Biological Control Agent ◽  
Control Agent ◽  
Aspergillus Species ◽  
Expression Data ◽  
Altered Gene Expression ◽  
Large Numbers ◽  
Altered Gene ◽  
Infected Line

This study determined the effects of Aspergillus thermomutatus chrysovirus 1 (AthCV1), isolated from Aspergillus thermomutatus, on A. fumigatus, A. nidulans and A. niger. Protoplasts of virus-free isolates of A. fumigatus, A. nidulans and A. niger were transfected with purified AthCV1 particles and the phenotype, growth and sporulation of the isogenic AthCV1-free and AthCV1-infected lines assessed at 20 °C and 37 °C and gene expression data collected at 37 °C. AthCV1-free and AthCV1-infected A. fumigatus produced only conidia at both temperatures but more than ten-fold reduced compared to the AthCV1-infected line. Conidiation was also significantly reduced in infected lines of A. nidulans and A. niger at 37 °C. AthCV1-infected lines of A. thermomutatus and A. nidulans produced large numbers of ascospores at both temperatures, whereas the AthCV1-free line of the former did not produce ascospores. AthCV1-infected lines of all species developed sectoring phenotypes with sclerotia produced in aconidial sectors of A. niger at 37 °C. AthCV1 was detected in 18% of sclerotia produced by AthCV1-infected A. niger and 31% of ascospores from AthCV1-infected A. nidulans. Transcriptome analysis of the naturally AthCV1-infected A. thermomutatus and the three AthCV1-transfected Aspergillus species showed altered gene expression as a result of AthCV1-infection. The results demonstrate that AthCV1 can infect a range of Aspergillus species resulting in reduced sporulation, a potentially useful attribute for a biological control agent.

scholarly journals Aphid Honeydew Enhances Parasitoid Longevity to the Same Extent as a High-Quality Floral Resource: Implications for Conservation Biological Control of the Wheat Stem Sawfly (Hymenoptera: Cephidae)

2020 ◽  
Vol 113 (4) ◽  
pp. 2022-2025
Author(s):  
Tatyana A Rand ◽  
Debra K Waters
Keyword(s):  
Biological Control ◽  
Cover Crop ◽  
Habitat Management ◽  
Biological Control Agent ◽  
Parasitoid Wasp ◽  
Floral Resources ◽  
Cereal Aphid ◽  
Crop Species ◽  
Wheat Stem Sawfly ◽  
Aphid Honeydew

Abstract Providing sugar resources for parasitoids is an important component of habitat management approaches to bolster biological control. We screened three flowering cover crop species, and one aphid species, for their potential to increase the longevity of the parasitoid wasp, Bracon cephi (Gahan) (Hymenoptera: Braconidae), an important biological control agent of the wheat stem sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae). We found that buckwheat and honeydew from the cereal aphid, Rhopalosiphum padi (Linnaeus) (Hemiptera: Aphididae), increased longevity of B. cephi females by over threefold, while longevity on sunflower and coriander was not significantly different from controls on wheat. The results suggest that incorporating buckwheat into cover crop mixes could enhance parasitoid performance. However, the finding that honeydew associated with a common aphid in wheat provides a suitable resource suggests that a better understanding of the varying quality, and spatial and temporal availability, of aphid honeydew will be a critical consideration in evaluating the potential benefits of managing floral resources for parasitoid conservation in this system.

Evaluation of Appropriate Reference Genes For Investigating Gene Expression in Chlorops oryzae (Diptera: Chloropidae)

2019 ◽  
Vol 112 (5) ◽  
pp. 2207-2214 ◽  
Author(s):  
Ping Tian ◽  
Lin Qiu ◽  
Ailin Zhou ◽  
Guo Chen ◽  
Hualiang He ◽  
...  
Keyword(s):  
Gene Expression ◽  
Ribosomal Protein ◽  
Reference Genes ◽  
Molecular Mechanisms ◽  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Housekeeping Genes ◽  
Future Research ◽  
Economic Damage ◽  
Physiological Processes

Abstract Reverse transcription quantitative polymerase chain reaction (PCR) has become an invaluable technique for analyzing gene expression in many insects. However, this approach requires the use of stable reference genes to normalize the data. Chlorops oryzae causes significant economic damage to rice crops throughout Asia. The lack of suitable reference genes has hindered research on the molecular mechanisms underlying many physiological processes of this species. In this study, we used quantitative real-time PCR to evaluate the expression of eight C. oryzae housekeeping genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-actin (βACT), beta-tubulin (βTUB), Delta Elongation factor-1 (EF1δ), ribosomal protein S11 (RPS11), RPS15, C-terminal-Binding Protein (CtBP), and ribosomal protein 49 (RP49) in different developmental stages and tissues in both larvae and adults. We analyzed the data with four different software packages: geNorm, NormFinder, BestKeeper, and RefFinder and compared the results obtained with each method. The results indicate that PRS15 and RP49 can be used as stable reference genes for quantifying gene expression in different developmental stages and larval tissues. GAPDH and βACT, which have been considered stable reference genes by previous studies, were the least stable of the candidate genes with respect to larval tissues. GAPDH was, however, the most stable reference gene for adult tissues. We verified the candidate reference genes identified and found that the expression levels of Cadherins (Cads) changed when different reference genes were used to normalize gene expression. This study provides a valuable foundation for future research on gene function, and investigating the molecular basis of physiological processes, in C. oryzae.

scholarly journals Identification and Validation of Novel Reference Genes in Acute Lymphoblastic Leukemia for Droplet Digital PCR

Genes ◽  
2019 ◽  
Vol 10 (5) ◽  
pp. 376 ◽  
Author(s):  
Vanessa Villegas-Ruíz ◽  
Karina Olmos-Valdez ◽  
Kattia Alejandra Castro-López ◽  
Victoria Estefanía Saucedo-Tepanecatl ◽  
Josselen Carina Ramírez-Chiquito ◽  
...  
Keyword(s):  
Gene Expression ◽  
Acute Lymphoblastic Leukemia ◽  
Reference Genes ◽  
Expression Profiles ◽  
Lymphoblastic Leukemia ◽  
Gene Expression Profiles ◽  
Housekeeping Genes ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Cellular Processes

Droplet digital PCR is the most robust method for absolute nucleic acid quantification. However, RNA is a very versatile molecule and its abundance is tissue-dependent. RNA quantification is dependent on a reference control to estimate the abundance. Additionally, in cancer, many cellular processes are deregulated which consequently affects the gene expression profiles. In this work, we performed microarray data mining of different childhood cancers and healthy controls. We selected four genes that showed no gene expression variations (PSMB6, PGGT1B, UBQLN2 and UQCR2) and four classical reference genes (ACTB, GAPDH, RPL4 and RPS18). Gene expression was validated in 40 acute lymphoblastic leukemia samples by means of droplet digital PCR. We observed that PSMB6, PGGT1B, UBQLN2 and UQCR2 were expressed ~100 times less than ACTB, GAPDH, RPL4 and RPS18. However, we observed excellent correlations among the new reference genes (p < 0.0001). We propose that PSMB6, PGGT1B, UBQLN2 and UQCR2 are housekeeping genes with low expression in childhood cancer.

Sign in / Sign up

Export Citation Format

Share Document