US EPA

1173942 

Journal Article 

Rapid detection of plasmodesmata in purified cell walls 

Ritzenthaler, C; Findlay, K; Roberts, K; Maule, AJ 

2000 

1 

Protoplasma
ISSN: 0033-183X
EISSN: 1615-6102 

211 

3-4 

165-171 

WOS:000086996800005 

Plasmodesmata are complex channels within the plant cell wall, which create plasma membrane and symplastic continuity between neighbouring cells. To detect plasmodesmata in cell wall preparations from Nicotiana de elandii, we have used 3,3'-dihexyloxacarbocyanine iodide (DiOC(6)), a cationic amphiphilic fluorescent probe, widely employed for general studies of membrane structure and dynamics. Punctate fluorescent staining was readily seen in pit fields, small depressions within the cell wall known to be rich in plasmodesmata. Scanning electron microscopy was used to demonstrate that the punctate staining corresponded to plasmodesmata. Treatment of cell wail fragments with chloroform-methanol to remove lipids did not alter the staining of plasmodesmata. In contrast, pronase E-sodium dodecyl sulfate treatment completely abolished staining, indicating that the DiOC(6) labelling of plasmodesmata may be protein rather than lipid specific. Although not membrane mediated, DiOC(6) staining of plasmodesmata is a simple, rapid, and specific tool for the detection of plasmodesmata in isolated cell walls and will prove useful for studies of plasmodesmal location, structure, and composition. 

3,3 '-dihexyloxacarbocyanine iodide; plasmodesma; pit field; Nicotiana cle elandii