Methodology of freeze-substitution for study of freezing injury in plant cells by transmission electron microscopy

Cryobiology ◽  
1988 ◽  
Vol 25 (6) ◽  
pp. 528 ◽  
Author(s):  
D.M.R. Harvey ◽  
K. Pihakaski
Author(s):  
E. R. Rivera

Mature plants of Ferocactus latispinus were divided into the apical meristem, cortical and cambial tissue, and young and mature photosynthetic tissue. Pieces 2x2x4 mm were fixed in 0.075 M PIPES buffered 5% glutaraldehyde and further treated for conventional transmission electron microscopy.The shoot meristematic cells contained dense cytoplasm with a few small vacuoles. All organelles expected to be found in unspecialized plant cells were present. The proplastids were 1-2 diameters larger than mitochondria. Some of these plastids were elongated and most contained small amounts of lipid globules in the stroma (Fig. 1). In addition, phytoferritin was sometimes found in the stroma. No starch was found in these organelles. The thylakoids were poorly developed, and when there were no inclusions in the stroma, proplastids were difficult to distinguish from mitochondria (Fig. 1).Cortical cells had very large, well-developed vacuoles. The organelles were restricted to the peripheral cytoplasm of the cell and to a few transvacuolar cytoplasmic strands.


2011 ◽  
Vol 17 (S2) ◽  
pp. 162-163
Author(s):  
E Ebong ◽  
F Macaluso ◽  
D Spray ◽  
J Tarbell

Extended abstract of a paper presented at Microscopy and Microanalysis 2011 in Nashville, Tennessee, USA, August 7–August 11, 2011.


2001 ◽  
Vol 79 (1) ◽  
pp. 49-57 ◽  
Author(s):  
C W Mims ◽  
C Rodriguez-Lother ◽  
E A Richardson

A combination of scanning and transmission electron microscopy was used to examine the host-pathogen relationship in leaves of Duchesnea indica (Andrz) Focke infected by the rust fungus Frommeëla mexicana var. indicae McCain & Hennen. Samples for transmission electron microscopy were prepared using high pressure freezing followed by freeze substitution. This protocol provided excellent preservation of both host cells and fungal haustoria. Each haustorium of F. mexicana var. indicae possessed a long slender neck with a neck band and an expanded body that contained two nuclei positioned close together. The haustorial body was lobed and sometimes even branched but lacked septa. Details of the extrahaustorial membrane that separated each haustorium from the cytoplasm of its host cell were particularly well preserved. Extensive labyrinth cell wall ingrowths developed around haustorial necks, as well as elsewhere, in infected cells. These ingrowths appeared to be identical to those present in plant transfer cells. Transfer cells are thought to be involved in intensive solute transfer over short distances. This appears to be the first report of the development of transfer cells in response to infection by a plant pathogenic fungus.Key words: haustoria, transfer cells, freeze substitution, electron microscopy.


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