Isolation of Intact and Functional Chloroplasts from Mesophyll and Bundle Sheath Protoplasts of the C4 Plant Panicum miliaceum1
AUTOR(ES)
Edwards, Gerald E.
RESUMO
A procedure is described for isolating and purifying mesophyll protoplasts and bundle sheath protoplasts of the C4 plant Panicum miliaceum. Following enzymic digestion of leaf tissue, mesophyll protoplasts and bundle sheath protoplasts are released and purified by density centrifugation. The lower density of mesophyll protoplasts allowed rapid separation of the two protoplast types. Evidence for separation of mesophyll protoplasts and bundle sheath protoplasts (up to 95% purity) is provided from light microscopy (based on size difference in both chloroplasts and protoplasts), levels of marker enzymes in the preparations (i.e. pyruvate, Pi dikinase and phosphoenolpyruvate carboxylase for mesophyll and ribulose-1,5-bisphosphate carboxylase for bundle sheath), and differences in substrate-dependent O2 evolution by chloroplasts isolated from protoplasts.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=542927Documentos Relacionados
- Fluorescence and Delayed Light Emission from Mesophyll and Bundle Sheath Cells in Leaves of Normal and Salt-Treated Panicum miliaceum1
- Photophosphorylation by Mesophyll and Bundle Sheath Chloroplasts of C4 Plants
- Metabolic Activities in Extracts of Mesophyll and Bundle Sheath Cells of Panicum miliaceum (L.) in Relation to the C4 Dicarboxylic Acid Pathway of Photosynthesis 1
- Distribution of Nitrate-assimilating Enzymes between Mesophyll Protoplasts and Bundle Sheath Cells in Leaves of Three Groups of C4 Plants 1
- Use of Silica Sol Step Gradients to Prepare Bundle Sheath and Mesophyll Chloroplasts from Panicum maximum1