China Agricultural University 9 articles published in JoVE Behavior An Adjustable High-Definition Imaging System for Behavioral Studies of Drosophila Adults Tong Li*1,2, Yujia Weng*1,2, Daxiang Yang1 1Department of Zoology and Animal Physiology, College of Biological Sciences, China Agricultural University, 2College of Life Sciences, Capital Normal University This protocol describes how to make a simple adult Drosophila behavior observation chamber, and how to take high-definition photographs/videos of the morphology or behavior of different types of fruit flies in the observation chamber through relatively simple and affordable methods. Immunology and Infection TurboID-Based Proximity Labeling for In Planta Identification of Protein-Protein Interaction Networks Yongliang Zhang*1, Yuanyuan Li*2,3, Xinxin Yang1, Zhiyan Wen1, Ugrappa Nagalakshmi2, Savithramma P. Dinesh-Kumar2,3 1State Key Laboratory of Agro-Biotechnology and Ministry of Agriculture Key Laboratory of Soil Microbiology, College of Biological Sciences, China Agricultural University, 2Department of Plant Biology, College of Biological Sciences, University of California, Davis, 3Genome Center, College of Biological Sciences, University of California, Davis Described here is a proximity labeling method for identification of interaction partners of the TIR domain of the NLR immune receptor in Nicotiana benthamiana leaf tissue. Also provided is a detailed protocol for the identification of interactions between other proteins of interest using this technique in Nicotiana and other plant species. Biology Nucleofection and In Vivo Propagation of Chicken Eimeria Parasites Chunhui Duan1, Xinming Tang1, Dandan Hu1, Sixin Zhang1, Jie Liu1, Feifei Bi1, Zhenkai Hao1, Jingxia Suo1, Ying Yu1, Min Wang1, Pei Sun1, Liying Du2, Xun Suo1, Xianyong Liu1 1National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, 2School of Life Sciences, Peking University Here, we provided a method to achieve stable transfection of chicken Eimeria parasites by nucleofecting sporozoites or second-generation merozoites. Genetically modified eimerian parasites expressing heterologous antigenic genes could be used as vaccine delivery vehicles. Biology Simple Homemade Tools to Handle Fruit Flies—Drosophila melanogaster Daxiang Yang1 1Department of Zoology and Animal Physiology, College of Biological Sciences, China Agricultural University Described here is the use of several homemade tools to transfer, chill, and kill adult Drosophila, as well as to clean glass culture vials and collect eggs. These tools are easy to make and are rather efficient in handling Drosophila. Immunology and Infection Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice Yongli Zhang*1,2, Hangjie Zhang*2, Wenqiang Ma*3, Kefang Liu1,2, Min Zhao4, Yingze Zhao2, Xuancheng Lu5, Fuping Zhang6, Xiangdong Li3, George F. Gao1,2,4,6, William J. Liu1,2 1School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, 2NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 3State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, 4Research Network of Immunity and Health (RNIH), Beijing Institutes of Life Science, Chinese Academy of Sciences, 5Laboratory Animal Center, Chinese Center for Disease Control and Prevention, 6CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences A protocol to evaluate antigen-specific T-cell responses in the immunoprivileged organs of the Ifnar1-/- murine model for the Zika virus (ZIKV) infection is described. This method is pivotal for investigating the cellular mechanisms of the protection and immunopathogenesis of ZIKV vaccines and is also valuable for their efficacy evaluation. Environment The Plant Infection Test: Spray and Wound-Mediated Inoculation with the Plant Pathogen Magnaporthe Grisea Mengyu Zhang*1, Xuan Sun*1, Lie Cui1, Yue Yin1, Xinyu Zhao1, Song Pan2, Weixiang Wang1 1Beijing Key Laboratory of New Technology in Agricultural Application, National Demonstration Center for Experimental Plant Production Education, Beijing University of Agriculture, 2College of Plant Protection, State Key Laboratory of Agrobiotechnology, Ministry of Agriculture Key Laboratory of Pest Monitoring and Green Management, China Agricultural University Here, we present a protocol to test plant virulence with the plant pathogen Magnaporthe grisea. This report will contribute to the large-scale screening of the pathotypes of fungi isolates and serve as an excellent starting point for understanding the resistant mechanisms of plants during molecular breeding. Neuroscience Single Sensillum Recordings for Locust Palp Sensilla Basiconica Hongwei Li1, Yinwei You2, Long Zhang1 1Department of Entomology, China Agricultural University, 2Bio-tech Research Center, Shandong Academy of Agricultural Sciences This paper describes a detailed and highly effective protocol for single sensillum recordings from the sensilla basiconica on the palps of insect mouthparts. Neuroscience Localization of Odorant Receptor Genes in Locust Antennae by RNA In Situ Hybridization Xiao Xu1, Yinwei You1,2, Long Zhang1 1Department of Entomology, China Agricultural University, 2Bio-tech Research Center, Shandong Academy of Agricultural Sciences This paper describes a detailed and highly effective RNA in situ hybridization protocol particularly for low-level expressed Odorant Receptor (OR) genes, as well as other genes, in insect antennae using digoxigenin (DIG)-labeled or biotin-labeled probes. Medicine Apical Resection Mouse Model to Study Early Mammalian Heart Regeneration Jianhua Xiong1, Jian Hou2 1Center for Molecular Medicine, National Heart, Lung, and Blood Institute, National Institutes of Health, 2State Key Laboratory for Agrobiotechnology, College of Biological Science, China Agricultural University A step-by-step video protocol of apical resection is demonstrated in this study. Apical resection is a recently highlighted surgical approach in mammalian heart regeneration research. This study may promote the application of apical resection as a standard methodology in research into the mechanism underlying heart regeneration.