Ultrastructural and functional similarity of the haustorial neckband of rust fungi and the Casparian strip of vascular plants

1976 ◽  
Vol 54 (21) ◽  
pp. 2484-2489 ◽  
Author(s):  
Michèle C. Heath

In susceptible hosts infected with rust fungi, an osmiophilic band, bridging the interface between host and parasite, is usually observed about midway along the haustorial neck. This neckband bears certain ultrastructural resemblances to the Casparian strip of endodermal cells, and the possible similarity in function as a barrier to apoplastic transport was investigated using 4-acetamido, 4′-isothiocyanostilbene-2, 2′-disulphonic acid (SITS) and uranyl acetate solutions. Both these reagents appeared to travel along host and fungal walls but did not readily penetrate the protoplasts of either organism.When rust-infected cowpea leaves were infiltrated with SITS reagent, haustoria in broken cells fluoresced as brightly as the walls of both intercellular hyphae and host cells. Mature haustoria in intact cells, however, did not fluoresce, suggesting the presence of a barrier to the flow of reagent to the haustorium. When rusted corn leaves were allowed to take up uranyl acetate solution, the ultrastructural distribution of uranyl crystals indicated that this barrier was, indeed, the neckband, and additional circumstantial evidence for the function of this structure was provided by indications from both techniques that the transport barrier was absent from very young haustoria; the latter have previously been shown to lack an ultrastructurally detectable neckband during the early stages of their development.These results appear to confirm the functional, as well as morphological, similarity of the haustorial neckband and the Casparian strip, and these, therefore, present an unusual example of parallel evolution at the structural and functional level. The importance of such a barrier to apoplastic transport in haustoria-producing fungi is discussed.

Author(s):  
H.A. Cohen ◽  
W. Chiu ◽  
J. Hosoda

GP 32 (molecular weight 35000) is a T4 bacteriophage protein that destabilizes the DNA helix. The fragment GP32*I (77% of the total weight), which destabilizes helices better than does the parent molecule, crystallizes as platelets thin enough for electron diffraction and electron imaging. In this paper we discuss the structure of this protein as revealed in images reconstructed from stained and unstained crystals.Crystals were prepared as previously described. Crystals for electron microscopy were pelleted from the buffer suspension, washed in distilled water, and resuspended in 1% glucose. Two lambda droplets were placed on grids over freshly evaporated carbon, allowed to sit for five minutes, and then were drained. Stained crystals were prepared the same way, except that prior to draining the droplet, two lambda of aqueous 1% uranyl acetate solution were applied for 20 seconds. Micrographs were produced using less than 2 e/Å2 for unstained crystals or less than 8 e/Å2 for stained crystals.


Author(s):  
O. H. Kapp ◽  
M. Ohtsuki ◽  
N. Robin ◽  
S. N. Vinogradov ◽  
A. V. Crewe

Annelid extracellular hemoglobins are among the largest known proteins (M.W = 3.9 x 106), and together with the hemocyanins are the largest known oxygen carriers. They display oxygen affinities generally higher than those o vertebrate hemoglobins with Hill coefficients ranging from slightly higher than unity to values as high as 5-6. These complex molecules are composed of multiple copies of as many as six different polypeptides and posse: approximately 150 hemes per molecule.The samples were diluted to 100-200 μg/ml with distilled water just before application to a thin carbon film (∽15 Å thick). One percent (w/v) uranyl acetate solution was used for negative staining for 2 minutes and dried in air. The specimens were examined with the high resolution STEM. Their general appearance is that of a hexagonal bilayer (Fig. 1), each layer consisting of six spheroidal subunits. The corner to corner hexagonal dimensic is approximately 300 Å and the bilayer thickness approximately 200 Å.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Guilhem Reyt ◽  
Priya Ramakrishna ◽  
Isai Salas-González ◽  
Satoshi Fujita ◽  
Ashley Love ◽  
...  

AbstractLignin is a complex polymer deposited in the cell wall of specialised plant cells, where it provides essential cellular functions. Plants coordinate timing, location, abundance and composition of lignin deposition in response to endogenous and exogenous cues. In roots, a fine band of lignin, the Casparian strip encircles endodermal cells. This forms an extracellular barrier to solutes and water and plays a critical role in maintaining nutrient homeostasis. A signalling pathway senses the integrity of this diffusion barrier and can induce over-lignification to compensate for barrier defects. Here, we report that activation of this endodermal sensing mechanism triggers a transcriptional reprogramming strongly inducing the phenylpropanoid pathway and immune signaling. This leads to deposition of compensatory lignin that is chemically distinct from Casparian strip lignin. We also report that a complete loss of endodermal lignification drastically impacts mineral nutrients homeostasis and plant growth.


2018 ◽  
Author(s):  
Debnath Ghosal ◽  
Yi-Wei Chang ◽  
Kwang Cheol Jeong ◽  
Joseph P. Vogel ◽  
Grant J. Jensen

AbstractLegionella pneumophilasurvives and replicates inside host cells by secreting ~300 effectors through the Dot/Icm type IVB secretion system (T4BSS). Understanding this machine’s structure is challenging because of its large number of components (27) and integration into all layers of the cell envelope. Previously we overcame this obstacle by imaging the Dot/Icm T4BSS in its native state within intact cells through electron cryotomography. Here we extend our observations by imaging a stabilized mutant that yielded a higher resolution map. We describe for the first time the presence of a well-ordered central channel that opens up into a windowed large (~32 nm wide) secretion chamber with an unusual 13-fold symmetry. We then dissect the complex by matching proteins to densities for many components, including all those with periplasmic domains. The placement of known and predicted structures of individual proteins into the map reveals the architecture of the T4BSS and provides a roadmap for further investigation of this amazing specialized secretion system.


1971 ◽  
Vol 49 (1) ◽  
pp. 35-38 ◽  
Author(s):  
E. B. Dumbroff ◽  
D. R. Peirson

The endodermis, with its associated Casparian strip, is generally believed to act as an effective barrier to the passive movement of ions from the cortex to the xylem in young roots. However, several workers have suggested that the functional integrity of the endodermis might be somewhat impaired with the emergence of branch roots from the pericycle, thus providing pathways for the mass flow of water and ions into the stele. The present work was undertaken to examine the validity of this hypothesis.Sections of lateral roots embedded in glycol methacrylate were stained and examined by fluorescence microscopy, and a general picture of the relationship between branch root development and concomitant changes in the endodermis emerged. The endodermal cells of the parent root were found to maintain a continuous, unbroken, suberized layer over the surface of a very young lateral root, but with continued elongation there is a period when formation of the Casparian strip lags behind division of endodermal cells. It appears likely that, at this stage, water and ions can enter the stele of the parent root by mass flow.


2008 ◽  
Vol 76 (4) ◽  
pp. 1476-1484 ◽  
Author(s):  
Anne A. Wolf ◽  
Michael G. Jobling ◽  
David E. Saslowsky ◽  
Eli Kern ◽  
Kimberly R. Drake ◽  
...  

ABSTRACT Cholera toxin (CT) moves from the plasma membrane (PM) of host cells to the endoplasmic reticulum (ER) by binding to the lipid raft ganglioside GM1. The homopentomeric B-subunit of the toxin can bind up to five GM1 molecules at once. Here, we examined the role of polyvalent binding of GM1 in CT action by producing chimeric CTs that had B-subunits with only one or two normal binding pockets for GM1. The chimeric toxins had attenuated affinity for binding to host cell PM, as expected. Nevertheless, like wild-type (wt) CT, the CT chimeras induced toxicity, fractionated with detergent-resistant membranes extracted from toxin-treated cells, displayed restricted diffusion in the plane of the PM in intact cells, and remained bound to GM1 when they were immunoprecipitated. Thus, binding normally to two or perhaps only one GM1 molecule is sufficient for association with lipid rafts in the PM and toxin action. The chimeric toxins, however, were much less potent than wt toxin, and they entered the cell by endocytosis more slowly, suggesting that clustering of GM1 molecules by the B-subunit enhances the efficiency of toxin uptake and perhaps also trafficking to the ER.


Planta ◽  
2001 ◽  
Vol 213 (3) ◽  
pp. 474-477 ◽  
Author(s):  
Masaki Yokoyama ◽  
Ichirou Karahara

1969 ◽  
Vol 47 (12) ◽  
pp. 1869-1871 ◽  
Author(s):  
D. R. Peirson ◽  
E. B. Dumbroff

A new combination of embedding material and high contrast stain has provided the means for demonstrating, photographically, tangential sections of endodermal cells showing complete Casparian strips.


2015 ◽  
Vol 112 (33) ◽  
pp. 10533-10538 ◽  
Author(s):  
Takehiro Kamiya ◽  
Monica Borghi ◽  
Peng Wang ◽  
John M. C. Danku ◽  
Lothar Kalmbach ◽  
...  

The endodermis in roots acts as a selectivity filter for nutrient and water transport essential for growth and development. This selectivity is enabled by the formation of lignin-based Casparian strips. Casparian strip formation is initiated by the localization of the Casparian strip domain proteins (CASPs) in the plasma membrane, at the site where the Casparian strip will form. Localized CASPs recruit Peroxidase 64 (PER64), a Respiratory Burst Oxidase Homolog F, and Enhanced Suberin 1 (ESB1), a dirigent-like protein, to assemble the lignin polymerization machinery. However, the factors that control both expression of the genes encoding this biosynthetic machinery and its localization to the Casparian strip formation site remain unknown. Here, we identify the transcription factor, MYB36, essential for Casparian strip formation. MYB36 directly and positively regulates the expression of the Casparian strip genes CASP1, PER64, and ESB1. Casparian strips are absent in plants lacking a functional MYB36 and are replaced by ectopic lignin-like material in the corners of endodermal cells. The barrier function of Casparian strips in these plants is also disrupted. Significantly, ectopic expression of MYB36 in the cortex is sufficient to reprogram these cells to start expressing CASP1–GFP, correctly localize the CASP1–GFP protein to form a Casparian strip domain, and deposit a Casparian strip-like structure in the cell wall at this location. These results demonstrate that MYB36 is controlling expression of the machinery required to locally polymerize lignin in a fine band in the cell wall for the formation of the Casparian strip.


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