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9 ARTICLES PUBLISHED IN JoVE

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Environment

Design and Operation of a Continuous 13C and 15N Labeling Chamber for Uniform or Differential, Metabolic and Structural, Plant Isotope Labeling
Jennifer L Soong 1, Dan Reuss 1, Colin Pinney 1, Ty Boyack 1, Michelle L Haddix 1, Catherine E Stewart 2, M. Francesca Cotrufo 1,3
1Natural Resource Ecology Laboratory, Colorado State University, 2Soil Plant Nutrient REsearch, USDA-ARS, 3Department of Soil and Crop Sciences, Colorado State University

This method explains how to build and operate a continuous 13C and 15N isotope labeling chamber for uniform or differential plant tissue labeling. Representative results from metabolic and structural labeling of Andropogon gerardii are discussed.

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Biology

Methods for Comparing Nutrients in Beebread Made by Africanized and European Honey Bees and the Effects on Hemolymph Protein Titers
Gloria Degrandi-Hoffman 1, Bruce Eckholm 2, Ming Huang 3
1Carl Hayden Bee Research Center, USDA-ARS, 2Coupeville, WA, USA, 3Ecotoxicology Division, Eurofins Agroscience Services, Inc.

Here are methods to quantify nutrients in pollen before and after its conversion to beebread by two subspecies of honeybees. We describe techniques to measure beebread consumption and resulting protein titers in both subspecies.

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JoVE Journal

Empirical, Metagenomic, and Computational Techniques Illuminate the Mechanisms by which Fungicides Compromise Bee Health
Shawn A. Steffan 1,2, Prarthana S. Dharampal 2, Luis Diaz-Garcia 3,4, Cameron R. Currie 5, Juan Zalapa 1,3, Chris Todd Hittinger 6,7,8
1Vegetable Crop Research Unit, USDA-ARS, 2Department of Entomology, University of Wisconsin-Madison, 3Department of Horticulture, University of Wisconsin-Madison, 4Instituto Nacional de Investigaciones Forestales, Agricolas y Pecuarias, 5Department of Bacteriology, University of Wisconsin-Madison, 6Laboratory of Genetics, Genome Center of Wisconsin, 7DOE Great Lakes Bioenergy Research Center, Wisconsin Energy Institute, 8J.F. Crow Institute for the Study of Evolution, University of Wisconsin-Madison

Microbial consortia within bumble bee hives enrich and preserve pollen for bee larvae. Using next generation sequencing, along with laboratory and field-based experiments, this manuscript describes protocols used to test the hypothesis that fungicide residues alter the pollen microbiome, and colony demographics, ultimately leading to colony loss.

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Environment

Procedures of Laboratory Fumigation for Pest Control with Nitric Oxide Gas
Yong-Biao Liu 1, Xiangbing Yang 2, Tiffany Masuda 1
1Crop Improvement and Protection Unit, USDA-ARS, 2The University of California Davis

This paper describes nitric oxide (NO) fumigation protocols for postharvest pest control. Fumigation chambers are flushed with nitrogen (N2) to establish ultralow oxygen conditions before NO is injected. At the end, chambers are flushed with N2 to dilute NO before exposing products to ambient air to prevent exposure to NO2.

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Environment

In Vitro Rearing of Solitary Bees: A Tool for Assessing Larval Risk Factors
Prarthana S. Dharampal 1, Caitlin M. Carlson 2, Luis Diaz-Garcia 3,4, Shawn A. Steffan 1,5
1Department of Entomology, University of Wisconsin-Madison, 2Department of Bacteriology, University of Wisconsin-Madison, 3Department of Horticulture, University of Wisconsin-Madison, 4Instituto Nacional de Investigaciones Forestales Agricolas y Pecuarias, 5USDA-ARS, Vegetable Crop Research Unit

Fungicide sprays on flowering plants may expose solitary bees to high concentrations of pollen-borne fungicide residues. Using laboratory-based experiments involving in vitro-reared bee larvae, this study investigates the interactive effects of consuming fungicide-treated pollen derived from host and non-host plants.

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Environment

Detached Leaf Assays to Simplify Gene Expression Studies in Potato During Infestation by Chewing Insect Manduca sexta
Nicole G. Novak 1, Frances G. Perez 2, Richard W. Jones 2, Susan D. Lawrence 1
1Invasive Insect Biocontrol and Behavior Lab, United States Department of Agriculture (USDA), Agricultural Research Service (ARS), 2Genetic Improvement for Fruits and Vegetables Lab, USDA-ARS

The presented method creates natural herbivore damaged plant tissue through the application of Manduca sexta larvae to detached leaves of potato. The plant tissue is assayed for expression of six transcription factor homologs involved in early responses to insect herbivory.

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Behavior

Using Flight Mills to Measure Flight Propensity and Performance of Western Corn Rootworm, Diabrotica virgifera virgifera (LeConte)
Eric Yu Yu 1, Aaron J. Gassmann 1, Thomas W. Sappington 2
1Department of Entomology, Iowa State University, 2Corn Insects & Crop Genetics Research Unit, USDA-ARS

Flight mills are important tools for comparing how age, sex, mating status, temperature, or various other factors may influence an insects flight behavior. Here we describe protocols to tether and measure the flight propensity and performance of western corn rootworm under different treatments.

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Genetics

A Pathway Association Study Tool for GWAS Analyses of Metabolic Pathway Information
Adam Thrash 1, Marilyn L Warburton 2
1Institute for Genomics, Biocomputing & Biotechnology, Mississippi State University, 2Corn Host Plant Resistance Research Unit, USDA-ARS

By running the Pathway Association Study Tool (PAST), either through the Shiny application or through the R console, researchers can gain a deeper understanding of the biological meaning of their genome-wide association study (GWAS) results by investigating the metabolic pathways involved.

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Biology

Assessing Agrochemical Risk to Mated Honey Bee Queens
Julia D. Fine 1, Kendall M. Torres 2, Jamilyn Martin 2, Gene E. Robinson 2,3,4
1Invasive Species and Pollinator Health Research Unit, USDA-ARS, 2Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 3Neuroscience Program, University of Illinois at Urbana-Champaign, 4Department of Entomology, University of Illinois at Urbana-Champaign

This protocol was developed to enhance the understanding of how agrochemicals affect honey bee (Apis mellifera) reproduction by establishing methods to expose honey bee queens and their worker caretakers to agrochemicals in a controlled, laboratory setting and carefully monitoring their relevant responses.

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