scholarly journals Variation in Expression of Reference Genes across Life Stages of a Bee, Megachile rotundata

Insects ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 36
Author(s):  
Junhuan Xu ◽  
Dennis L. Welker ◽  
Rosalind R. James
Keyword(s):  
Honey Bees ◽  
Reference Genes ◽  
Larval Growth ◽  
Environmental Toxins ◽  
Western United States ◽  
Life Stages ◽  
Megachile Rotundata ◽  
Alfalfa Seed ◽  
Transcription Data ◽  
Transcriptomic Studies

The alfalfa leafcutting bee, Megachile rotundata is widely used in the western United States as a pollinator for alfalfa seed production. Unfortunately, immatures experience high mortality in agriculturally managed populations. Quantified gene expression could be used to identify how this bee responds during different life stages to pathogens, environmental toxins, and other stresses, but stably expressed reference genes are needed to normalize transcription data. We evaluated twelve candidate genes for their transcription stability across different life stages, including during and after diapause. RPS18 and RPL8 were the two most stably expressed genes, followed by RPS5 and RPL27A. These genes were also very stable even during and after diapause, while the most variable genes being APN, PMIIM, NPC2, and Cr-PII had increased expression levels during larval growth and were also variable during and after diapause. The four reference genes we identified in M. rotundata may prove useful for transcriptomic studies in other bees as well, such as honey bees.

scholarly journals Antiviral Activities of a Medicinal Plant Extract Against Sacbrood Virus in Honeybees

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Liping Sun ◽  
Xueqi Zhang ◽  
Shufa Xu ◽  
Chunsheng Hou ◽  
Jin Xu ◽  
...  
Keyword(s):  
Honey Bees ◽  
Apis Cerana ◽  
Larval Growth ◽  
Antiviral Agent ◽  
Antiviral Effect ◽  
Absolute Quantification ◽  
Chinese Medicinal Herb ◽  
Sacbrood Virus ◽  
Antiviral Activities ◽  
Potential Strategy

Abstract Background Sacbrood is an infectious disease of the honey bee caused by Scbrood virus (SBV) which belongs to the family Iflaviridae and is especially lethal for Asian honeybee Apis cerana. Chinese Sacbrood virus (CSBV) is a geographic strain of SBV. Currently, there is a lack of an effective antiviral agent for controlling CSBV infection in honey bees. Methods Here, we explored the antiviral effect of a Chinese medicinal herb Radix isatidis on CSBV infection in A. cerana by inoculating the 3rd instar larvae with purified CSBV and treating the infected bee larvae with R. isatidis extract at the same time. The growth, development, and survival of larvae between the control and treatment groups were compared. The CSBV copy number at the 4th instar, 5th instar, and 6th instar larvae was measured by the absolute quantification PCR method. Results Bioassays revealed that R. isatidis extract significantly inhibited the replication of CSBV, mitigated the impacts of CSBV on larval growth and development, reduced the mortality of CSBV-infected A. cerana larvae, and modulated the expression of immune transcripts in infected bees. Conclusion Although the mechanism underlying the inhibition of CSBV replication by the medicine plant will require further investigation, this study demonstrated the antiviral activity of R. isatidis extract and provides a potential strategy for controlling SBV infection in honey bees.

EFFECT OF TWO INSECT VIRUSES AND TWO ENTOMOPATHOGENIC FUNGI ON LARVAL AND PUPAL DEVELOPMENT IN THE ALFALFA LEAFCUTTING BEE, MEGACHILE ROTUNDATA (FAB.) (HYMENOPTERA: MEGACHILIDAE)

1990 ◽  
Vol 122 (5) ◽  
pp. 1039-1040 ◽  
Author(s):  
D.W. Goerzen ◽  
M.A. Erlandson ◽  
K.C. Moore
Keyword(s):  
Apis Mellifera ◽  
Insect Pest ◽  
Microbial Control ◽  
Western Canada ◽  
Pest Species ◽  
Megachile Rotundata ◽  
Pupal Development ◽  
Canadian Prairies ◽  
Alfalfa Seed ◽  
Beneficial Insect

The alfalfa leafcutting bee, Megachile rotundata (Fab.), is an important pollinator of alfalfa in western Canada and is widely utilized for alfalfa seed production (Richards 1984). As a beneficial insect, the alfalfa leafcutting bee must be considered, along with the honey bee, Apis mellifera L., as a non-target organism of importance in evaluations of potential microbial control agents for use against major insect pest species on the Canadian prairies.

scholarly journals Reference gene analysis and its use for kinase expression profiling in Fasciola hepatica

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hicham Houhou ◽  
Oliver Puckelwaldt ◽  
Christina Strube ◽  
Simone Haeberlein
Keyword(s):  
Reference Genes ◽  
Drug Targets ◽  
Fasciola Hepatica ◽  
Expression Patterns ◽  
Housekeeping Gene ◽  
Trna Synthetase ◽  
Parasite Development ◽  
Mammalian Host ◽  
Life Stages

Abstract The liver fluke Fasciola hepatica causes fasciolosis, a foodborne zoonosis affecting humans and livestock worldwide. A reliable quantification of gene expression in all parasite life stages relevant for targeting by anthelmintics in the mammalian host is fundamental. The aim of this study was to define a set of stably expressed reference genes for qRT-PCR in Fasciola studies. We determined the expression stabilities of eight candidate reference genes by the algorithms NormFinder, geNorm, BestKeeper, and comparative ΔCT method. The most stably expressed reference genes for the comparison of intra-mammalian life stages were glutamyl-prolyl-tRNA synthetase (Fheprs) and tubulin-specific chaperone D (Fhtbcd). The two best reference genes for analysis of in vitro-cultured juveniles were Fhtbcd and proteasome subunit beta type-7 (Fhpsmb7). These genes should replace the housekeeping gene gapdh which is used in most Fasciola studies to date, but in fact was differentially expressed in our analysis. Based on the new reference genes, we quantified expression of five kinases (Abl1, Abl2, PKC, Akt1, Plk1) discussed as targets in other parasitic flatworms. Distinct expression patterns throughout development were revealed and point to interesting biological functions. We like to motivate using this set of validated reference genes for future F. hepatica research, such as studies on drug targets or parasite development.

scholarly journals Reference Genes for Addressing Gene Expression of Bladder Cancer Cell Models under Hypoxia: A Step Towards Transcriptomic Studies

PLoS ONE ◽  
2016 ◽  
Vol 11 (11) ◽  
pp. e0166120 ◽  
Author(s):  
Luís Lima ◽  
Cristiana Gaiteiro ◽  
Andreia Peixoto ◽  
Janine Soares ◽  
Manuel Neves ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bladder Cancer ◽  
Cancer Cell ◽  
Reference Genes ◽  
Bladder Cancer Cell ◽  
Cell Models ◽  
Transcriptomic Studies

Use of the freshwater macroinvertebrate Chironomus riparius (diptera: chironomidae) in the assessment of sediment toxicity

1996 ◽  
Vol 34 (7-8) ◽  
pp. 101-107 ◽  
Author(s):  
M. M. Watts ◽  
D. Pascoe
Keyword(s):  
Sediment Toxicity ◽  
Larval Growth ◽  
Sediment Contamination ◽  
Chironomus Riparius ◽  
Lower Percentage ◽  
Life Stages ◽  
Significant Growth ◽  
Polluted Sediment ◽  
Polluted Sites ◽  
Reference Sediment

The growth and emergence responses of the midge Chironomus riparius Meigen were used to evaluate the toxicity of natural and formulated reference sediments which were either obtained from polluted sites or spiked with copper in the laboratory. A significant reduction in larval growth was only observed in one of the two polluted field sediments tested. The reasons for this are discussed. However, copper spiking resulted in significant growth reductions of larvae exposed to the two formulated reference sediments at copper concentrations of 3 and 4 mg Cu2+ 1−1. Growth of larvae exposed in natural reference sediment was not significantly affected at equivalent concentrations. The emergence of adults was adversely affected in polluted field sediment compared to formulated reference sediment. Male, female and total adult median emergence times (EmT50) were significantly greater in the former treatment indicating a delay in emergence, attributable to sediment contamination. In addition, survivorship of the larvae in the polluted sediment was reduced, resulting in a lower percentage of adults emerging compared to the formulated reference sediment. The results of this study illustrate the usefulness of chronic response criteria, incorporating the more sensitive life-stages of C. riparius, in the assessment of sediment toxicity.

scholarly journals Presence of Pathogen-killed Larvae Influences Nesting Behavior of the Alfalfa Leafcutting Bee (Hymenoptera: Megachilidae)

2021 ◽  
Author(s):  
Ellen G Klinger ◽  
Dennis L Welker ◽  
Rosalind R James
Keyword(s):  
Natural Environment ◽  
Fungal Spores ◽  
Field Studies ◽  
Behavioral Changes ◽  
Nesting Behavior ◽  
Megachile Rotundata ◽  
Fungal Exposure ◽  
Nesting Sites ◽  
Alfalfa Seed ◽  
Nesting Patterns

Abstract The alfalfa leafcutting bee (Megachile rotundata (Fabricius)), a commercial pollinator used for alfalfa seed production, is susceptible to chalkbrood disease via ingested fungal spores. Diseases of insects can elicit behavioral changes in their hosts, but there are no recorded behaviors of alfalfa leafcutting bees in response to this fungal exposure. We conducted field studies to determine whether bees in pathogen-dense environments altered their nesting patterns, specifically if bees exposed to fungal spores produced higher numbers of nest cells and whether the proportions of nest cells that failed as eggs or small larvae (a state known as ‘pollen ball’) were greater. We found that our control bees, nontreated bees which were not exposed to chalkbrood spores other than those in the natural environment, had the highest proportion of pollen ball cells. Bees experimentally exposed to infective spores created the lowest number of nests and the fewest cells. Bees experimentally exposed to heat killed noninfective spores produced the greatest number of nests and cells overall and the greatest number of healthy progeny. We conclude that there are underlying behaviors that are elicited in response to the presence of chalkbrood spores that reduce the proportion of failed nest cells (grooming) and increase retention of bees at nesting sites (delay of bee emergence). Through further study of these behaviors, bee managers can potentially increase the productivity of their bee populations.

scholarly journals Validation of Novel Reference Genes for Reverse Transcription Quantitative Real-Time PCR in Drought-Stressed Sugarcane

2014 ◽  
Vol 2014 ◽  
pp. 1-12 ◽  
Author(s):  
Roberta Lane de Oliveira Silva ◽  
Manassés Daniel Silva ◽  
José Ribamar Costa Ferreira Neto ◽  
Claudia Huerta de Nardi ◽  
Sabrina Moutinho Chabregas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Reference Genes ◽  
Expression Profiles ◽  
Acc Oxidase ◽  
Quantitative Real Time Pcr ◽  
Experimental Conditions ◽  
Drought Tolerant ◽  
Statistical Approaches ◽  
Root Tissues ◽  
Transcriptomic Studies

One of the most challenging aspects of RT-qPCR data analysis is the identification of reliable reference genes. Ideally, they should be neither induced nor repressed under different experimental conditions. To date, few reference genes have been adequately studied for sugarcane (Saccharumspp.) using statistical approaches. In this work, six candidate genes (αTUB, GAPDH, H1, SAMDC, UBQ, and 25S rRNA) were tested for gene expression normalization of sugarcane root tissues from drought-tolerant and -sensitive accessions after continuous dehydration (24 h). By undergoing different approaches (GeNorm, NormFinder, and BestKeeper), it was shown that most of them could be used in combinations for normalization purposes, with the exception of SAMDC. Nevertheless three of them (H1,αTUB, and GAPDH) were considered the most reliable reference genes. Their suitability as reference genes validated the expression profiles of two targets (AS and PFPα1), related to SuperSAGE unitags, in agreement with results revealed by previousin silicoanalysis. The other two sugarcane unitags (ACC oxidase and PIP1-1), after salt stress (100 mM NaCl), presented their expressions validated in the same way. In conclusion, these reference genes will be useful for dissecting gene expression in sugarcane roots under abiotic stress, especially in transcriptomic studies using SuperSAGE or RNAseq approaches.

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