Preparation of RNA from Plant Tissue Using Guanidinium Isothiocyanate/Cesium Chloride Ultracentrifugation

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot4102
Author(s):  
Michael A. Connolly ◽  
Peter A. Clausen ◽  
James G. Lazar
Author(s):  
Russell L. Steere ◽  
Eric F. Erbe

It has been assumed by many involved in freeze-etch or freeze-fracture studies that it would be useless to etch specimens which were cryoprotected by more than 15% glycerol. We presumed that the amount of cryoprotective material exposed at the surface would serve as a contaminating layer and prevent the visualization of fine details. Recent unexpected freeze-etch results indicated that it would be useful to compare complementary replicas in which one-half of the frozen-fractured specimen would be shadowed and replicated immediately after fracturing whereas the complement would be etched at -98°C for 1 to 10 minutes before being shadowed and replicated.Standard complementary replica holders (Steere, 1973) with hinges removed were used for this study. Specimens consisting of unfixed virus-infected plant tissue infiltrated with 0.05 M phosphate buffer or distilled water were used without cryoprotectant. Some were permitted to settle through gradients to the desired concentrations of different cryoprotectants.


Author(s):  
Matias Pardo ◽  
Malcolm Slifkin ◽  
Leonard Merkow ◽  
Marie Sanchez

The simian adenoviruses SV20, SV30 and SA7 have been found to be oncogenic in the Syrian hamster. The growth characteristics and replicative cycle of these viruses in tissue culture therefore appeared appropriate to investigate. Cesium chloride purified simian adenovirus with an infectivity titer of 100 TCID50, was inoculated into monolayers of LLC-MK2 cells. Cells were fixed in osmium tetroxide and embedded for ultrastructural studies at 1, 3, 6, 9, 18, 24, 48, 72, 120 and 192 hours post-infection.At the first hour post-infection, virus particles were adsorbed to the plasmalemma and found within the peripheral cytoplasm of many LLC-MK2 cells (Fig. 1). Although the first detection of infectious virus occurred at 14 hours and infectivity titers did not reach a maximum until 30 hours, intranuclear virus particles were observed by 3 hours in typical adenovirus crystalline array (Fig. 2) by means of electron microscopy. These typical honeycomb arrayed virus particles at 3 hours provided evidence of significant replication in approximately 5 percent of tissue culture cells examined. Simultaneously, a classical nuclear inclusion manifested by peripheral condensation of nuclear chromatin was evident by light microscopy. As early at 6 to 9 hours, unusual intranuclear concentric membranes formed “tubes” which contained linear arranged virus particles (Fig. 3). In transverse or tangential sections, these “tubes” appeared cochlear-like in shape. In longitudinal section, these intranuclear tubular structures contained individual virus particles at various stages of maturation in a linear arranged order. This arrangement resembled “peas in a pod”.


Author(s):  
R. D. Sjolund ◽  
C. Y. Shih

The differentiation of phloem in plant tissue cultures offers a unique opportunity to study the development and structure of sieve elements in a manner that avoids the injury responses associated with the processing of similar elements in intact plants. Short segments of sieve elements formed in tissue cultures can be fixed intact while the longer strands occuring in whole plants must be cut into shorter lengths before processing. While iyuch controversy surrounds the question of phloem function in tissue cultures , sieve elements formed in these cultured cells are structurally similar to those of Intact plants. We are particullarly Interested In the structure of the plasma membrane and the peripheral ER in these cells because of their possible role in the energy-dependent active transport of sucrose into the sieve elements.


Planta Medica ◽  
2013 ◽  
Vol 79 (05) ◽  
Author(s):  
M Mujeeb ◽  
M Amir ◽  
AS Nadeem ◽  
M Aqil ◽  
AK Najmi ◽  
...  

Biomics ◽  
2019 ◽  
Vol 10 (4) ◽  
pp. 387-399 ◽  
Author(s):  
S.V. Veselova ◽  
G.F. Burkhanova ◽  
T.V. Nuzhnaya ◽  
S.D. Rumyantsev ◽  
I.V. Maksimov

2016 ◽  
Vol 128 (2) ◽  
pp. 437-446 ◽  
Author(s):  
Siham Esserti ◽  
Mohamed Faize ◽  
Lalla Aicha Rifai ◽  
Amal Smaili ◽  
Malika Belfaiza ◽  
...  

2016 ◽  
Vol 47 (5) ◽  
pp. 630-638 ◽  
Author(s):  
Elena Mikhailova ◽  
Donald Hagan ◽  
Julia Sharp ◽  
Tristan Allerton ◽  
Kylie Burdette ◽  
...  

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