Jiangsu Academy of Agricultural Sciences

6 ARTICLES PUBLISHED IN JoVE

Biology

Measuring Spatial and Temporal Ca²⁺ Signals in Arabidopsis Plants

Xiaohong Zhu¹, Aaron Taylor², Shenyu Zhang¹, Dayong Zhang^1,3, Ying Feng^1,4, Gaimei Liang^1,5, Jian-Kang Zhu^1,6

¹Department of Horticulture and Landscape Architecture, Purdue University, ²Bindley Bioscience Center, Purdue University, ³Institute of Biotechnology, Jiangsu Academy of Agricultural Sciences, ⁴College of Environmental & Resource Science, Zhejiang University, ⁵Dryland Agriculture Research Centre, Shanxi Academy of Agricultural Sciences, ⁶Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences

Ca²⁺ signaling regulates diverse biological processes in plants. Here we present approaches for monitoring abiotic stress induced spatial and temporal Ca²⁺ signals in Arabidopsis cells and tissues using the genetically encoded Ca²⁺ indicators Aequorin or Case12.

Environment

Specific and Accurate Detection of the Citrus Greening Pathogen Candidatus liberibacter spp. Using Conventional PCR on Citrus Leaf Tissue Samples

Huan Chen^*1,2, Ian Arthur Palmer^*1, Jian Chen^1,2, Ming Chang^1,2, Stephen L. Thompson³, Fengquan Liu², Zheng Qing Fu¹

¹Department of Biological Sciences, University of South Carolina, ²Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, ³Department of Instructions and Teacher Education, University of South Carolina

Citrus Greening is a particularly destructive disease affecting citrus crops globally. Presented here is a simple method using PCR and genomic DNA extraction of citrus leaf tissue for the accurate and precise identification of the citrus greening pathogen, Candidatus liberibacter spp.

Cancer Research

Patient-derived Heterogeneous Xenograft Model of Pancreatic Cancer Using Zebrafish Larvae as Hosts for Comparative Drug Assessment

Lei Wang^*1, Huan Chen^*2,3, Fei Fei^*1, Xianfeng He^2,3, Shaoyang Sun¹, Kunpeng Lv¹, Bo Yu^2,3, Jiang Long^2,4,5,6, Xu Wang¹

¹Department of Biochemistry and Molecular Biology, Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, School of Basic Medical Sciences, Fudan University, ²National Human Genetic Resources Sharing Service Platform (2005DKA21300), ³Fudan University Shanghai Cancer Center, ⁴Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, ⁵Department of Oncology, Shanghai Medical College, Fudan University, ⁶Pancreatic Cancer Institute, Fudan University

This protocol describes optimization procedures in a virus-based dual fluorescence-labeled tumor xenograft model using larval zebrafish as hosts. This heterogeneous xenograft model mimics the tissue composition of pancreatic cancer microenvironment in vivo and serves as a more precise tool for assessing drug responses in personalized zPDX (zebrafish patient-derived xenograft) models.

Genetics

CRISPR/Cas9 Ribonucleoprotein-mediated Precise Gene Editing by Tube Electroporation

Linyuan Ma^*1, Lydia Jang^*1, Jian Chen², Jun Song¹, Dongshan Yang¹, Jifeng Zhang¹, Y. Eugene Chen¹, Jie Xu¹

¹Center for Advanced Models for Translational Sciences and Therapeutics, Department of Internal Medicine, University of Michigan Medical Center, ²Celetrix LLC

Presented here is a protocol for efficient CRISPR/Cas9 ribonucleoprotein-mediated gene editing in mammalian cells using tube electroporation.

Medicine

A Training and Testing System for Performing Vascular Reconstruction In Vitro

Yue Wang^1,2, Lijun Mu³, Wei Zhang^1,2, Huan Chen^1,2, Qingshan Li^1,2, Aihua Shi¹, Bo Tang², Xiaogang Zhang^1,2, Dinghui Dong^1,2, Yi Lv^1,2

¹National Local Joint Engineering Research Center for Precision Surgery and Regenerative Medicine, First Affiliated Hospital of Xi'an Jiaotong University, ²Department of Hepatobiliary Surgery, First Affiliated Hospital of Xi'an Jiaotong University, ³Department of General Surgery, First Affiliated Hospital of Xi'an Jiaotong University

Here we present a training and testing system where a trainee can complete manual vascular reconstruction in vitro individually using a magnetic anchoring technique. The system can also be used to test the quality of reconstruction.

Immunology and Infection

Use of Fimbrial Rod for F18ab Fimbriae⁺ STEC Colonization to Host Cells

Mingxu Zhou^*1,2, Qiangde Duan^*1,3, Yang Yang^1,3, Guoqiang Zhu^1,3

¹College of Veterinary Medicine, Yangzhou University, ²Institute of Veterinary Immunology & Engineering, Jiangsu Academy of Agricultural Sciences, ³Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses

Here we present a protocol to study the function of fimbriae in bacterial colonization.