Methods for the Ultrastructural Analysis of Plant Cells and Tissues

Author(s):  
William V. Dashek ◽  
John E. Mayfield
Author(s):  
I. Brent Heath

Detailed ultrastructural analysis of fungal mitotic systems and cytoplasmic microtubules might be expected to contribute to a number of areas of general interest in addition to the direct application to the organisms of study. These areas include possibly fundamental general mechanisms of mitosis; evolution of mitosis; phylogeny of organisms; mechanisms of organelle motility and positioning; characterization of cellular aspects of microtubule properties and polymerization control features. This communication is intended to outline our current research results relating to selected parts of the above questions.Mitosis in the oomycetes Saprolegnia and Thraustotheca has been described previously. These papers described simple kinetochores and showed that the kineto- chores could probably be used as markers for the poorly defined chromosomes. Kineto- chore counts from serially sectioned prophase mitotic nuclei show that kinetochore replication precedes centriole replication to yield a single hemispherical array containing approximately the 4 n number of kinetochore microtubules diverging from the centriole associated "pocket" region of the nuclear envelope (Fig. 1).


Author(s):  
G. M. Hutchins ◽  
J. S. Gardner

Cytokinins are plant hormones that play a large and incompletely understood role in the life-cycle of plants. The goal of this study was to determine what roles cytokinins play in the morphological development of wheat. To achieve any real success in altering the development and growth of wheat, the cytokinins must be applied directly to the apical meristem, or spike of the plant. It is in this region that the plant cells are actively undergoing mitosis. Kinetin and Zeatin were the two cytokinins chosen for this experiment. Kinetin is an artificial hormone that was originally extracted from old or heated DNA. Kinetin is easily made from the reaction of adenine and furfuryl alcohol. Zeatin is a naturally occurring hormone found in corn, wheat, and many other plants.Chinese Spring Wheat (Triticum aestivum L.) was used for this experiment. Prior to planting, the seeds were germinated in a moist environment for 72 hours.


Author(s):  
Gerald Rupp

The marine protozoan Allogromia sp, strain NF Lee extends an elaborate reticulopodial network (RN) which contains an elongate microtubule-(MT)-based cytoskeleton. The MTs are located primarily within cytoplasmic fibrils which are visible by light microscopy (LM) in highly flattened or “two dimensionalized” reticulopodia. It was shown previously that allogromiid RNs withdraw in response to hypertonic Mg2+-seawater. An ultrastructural analysis of this phenomenon indicated that large patches of paracrystalline (PC) material, composed of helical filament aggregates, form concomitant with a decrease in MT number. Similar large patches of PC aggregates are also found in juvenile Allogromia before they extend a RN, which disappear during RN formation. Finally, PC aggregates are occasionally seen near microtubules in normal untreated RNs. Thus there is circumstantial evidence to propose that PC aggregates in Allogromia represent an intermediate form of tubulin; however, more definitive biochemical or immunocytochemical data is not available.


Author(s):  
Ann Cleary

Microinjection of fluorescent probes into living plant cells reveals new aspects of cell structure and function. Microtubules and actin filaments are dynamic components of the cytoskeleton and are involved in cell growth, division and intracellular transport. To date, cytoskeletal probes used in microinjection studies have included rhodamine-phalloidin for labelling actin filaments and fluorescently labelled animal tubulin for incorporation into microtubules. From a recent study of Tradescantia stamen hair cells it appears that actin may have a role in defining the plane of cell division. Unlike microtubules, actin is present in the cell cortex and delimits the division site throughout mitosis. Herein, I shall describe actin, its arrangement and putative role in cell plate placement, in another material, living cells of Tradescantia leaf epidermis.The epidermis is peeled from the abaxial surface of young leaves usually without disruption to cytoplasmic streaming or cell division. The peel is stuck to the base of a well slide using 0.1% polyethylenimine and bathed in a solution of 1% mannitol +/− 1 mM probenecid.


Author(s):  
M. Yamada ◽  
K. Ueda ◽  
K. Kuboki ◽  
H. Matsushima ◽  
S. Joens

Use of variable Pressure SEMs is spreading among electron microscopists The variable Pressure SEM does not necessarily require specimen Preparation such as fixation, dehydration, coating, etc which have been required for conventional scanning electron microscopy. The variable Pressure SEM allows operating Pressure of 1˜270 Pa in specimen chamber It does not allow microscopy of water-containing specimens under a saturated vapor Pressure of water. Therefore, it may cause shrink or deformation of water-containing soft specimens such as plant cells due to evaporation of water. A solution to this Problem is to lower the specimen temperature and maintain saturated vapor Pressures of water at low as shown in Fig. 1 On this technique, there is a Published report of experiment to have sufficient signal to noise ratio for scondary electron imaging at a relatively long working distance using an environmental SEM. We report here a new low temperature microscopy of soft Plant cells using a variable Pressure SEM (Hitachi S-225ON).


1992 ◽  
Vol 2 (5) ◽  
pp. 809-813 ◽  
Author(s):  
K Gordon ◽  
J Futterer ◽  
T Hohn

Sign in / Sign up

Export Citation Format

Share Document