Work

Stomata Tape-Peel: An Improved Method for Guard Cell Sample Preparation

Year: 2018

Type: article

Abstract: The study of guard cells is essential to the knowledge of the specific contributions of this cell type to the overall function of plants. However, it is often difficult to isolate and thus studying them often provides a challenge. This study establishes a stomatal guard cell enrichment method. The protocol utilizes Scotch tape to isolate guard cells and extract proteins from the cells. This method improves the integrity and yield of guard cell during isolation and provides a reliable sample for the study of cell signaling processes and how they correlate to stomatal movement phenotypes. In this method, the plant leaves were partitioned between two portions of tape and peeled apart to remove the abaxial side. This protocol was applied to Arabidopsis leaf tissue to isolate guard cells. The cells were treated with fluorescein diacetate (FDA) to determine viability and with abscisic acid (ABA) to assess stomata movement in response to ABA. In conclusion, this protocol proves useful for preparing enriched stomatal guard cells and isolating proteins. The quality of the cells and proteins obtained here enable physiological and other biological studies.

Source: Journal of Visualized Experiments

DOI: 10.3791/57422

Authors: Sheldon Lawrence II, Quiying Pang, Wenwen Kong, Sixue Chen

Institutions: University of Florida, Northeast Forestry University

Language: English

Topic: Plant Stress Responses and Tolerance

Subfield: Plant Science

Field: Agricultural and Biological Sciences

Domain: Life Sciences

Open Access status: green

Funders: National Science Foundation, National Natural Science Foundation of China, China Scholarship Council

Awards: COLLABORATIVE RESEARCH: Metabolomic Characterization of Red Light and CO2 Signaling in Guard Cells and Mesophyll Cells, Collaborative Research: Redox and Metabolomic Regulatory Mechanisms Underlying Guard Cell ABA Signal Transduction, G8511312752, Collaborative Research: Redox Regulation of Protein Kinase Functions in Guard Cell Signaling