In vitro formation of haustoria of the cowpea rust fungus Uromyces vignae in the absence of a living plant cell. II. Electron microscopy

1990 ◽  
Vol 68 (2) ◽  
pp. 278-287 ◽  
Author(s):  
Michèle C. Heath

The ultrastructure of infection hyphae, haustorial mother cells, and haustoria of Uromyces vignae formed on collodion membranes is described after conventional preparation and after treatment with periodic acid – thiocarbonhydrazide – silver proteinate or periodic acid – chromic acid – phosphotungstic acid treatments. Infection hyphae and haustorial mother cells developed normally in vitro but lacked an outer wall layer present in older infections in the host plant. Haustorium formation in vitro was accompanied by the development of elaborations of the plasmalemma along the infection hypha side of the haustorial mother cell septum, but their contents did not show identical staining responses to the haustorial neck wall as reported for other rust fungus species. Haustorial necks, and to a lesser extent haustorial bodies, were coated with a fibrillar material, the staining characteristics of which were similar to material normally considered part of the extrahaustorial matrix in infected plants. The restriction of this material to the haustorium suggests that it may play an important role in the interaction between the haustorium and the plant cell.




1994 ◽  
Vol 72 (6) ◽  
pp. 808-817 ◽  
Author(s):  
Pierre DesRochers ◽  
G. B. Ouellette

An unknown fungus isolated from an elm branch and inhibitory against Ophiostoma ulmi in vitro is described as Phaeotheca dimorphospora sp.nov. This dematiaceous deuteromycete propagates by endoconidia released after exfoliation of chlamydospore outer wall, as in mother cells of the type species Phaeotheca fissurella. However, P. dimorphospora differs from the type species by producing hyaline secondary ameroconidia between the endoconidial masses. Other ameroconidia, similar to the secondary ameroconidia, are produced through the chlamydospore outer wall. The optimal growth temperature of P. dimorphospora is 23 °C, whereas it is 15.5 °C for the type species. On media containing a high dextrose concentration (30 g ∙ L−1), colonies of P. dimorphospora are gray and crustose and grow slowly, at least initially. Conversely, on media with a low dextrose concentration (5 or 10 g ∙ L−1) colonies have a faster growth rate and appear whitish or ivory and fluffy. Key words: Phaeotheca dimorphospora, diagnosis, inhibition, Ophiostoma ulmi, Dutch elm disease.



1988 ◽  
Vol 66 (4) ◽  
pp. 742-744 ◽  
Author(s):  
Michèle C. Heath ◽  
M. R. Bonde

To investigate the developmental relationship between haustoria and intracellular hyphae, fixed and cleared whole mounts of com leaves infected with the tropical corn rust fungus Physopella zeae (Mains) Cumm. & Ramachar were examined by interference contrast light microscopy. Haustoria were clearly distinguished from intracellular hyphae by their morphological differentiation into a filamentous body and a narrow neck. The latter was encircled by a thin neckband, which appeared to develop in young haustoria at the time when the haustorial body started to expand. Observations made at the colony margins suggested that the fungus grew into uninvaded epidermal tissue in the following sequence: (i) formation of a haustorium from a haustorial mother cell in an adjacent cell, (ii) formation of an intracellular hypha next to the haustorium by the same intracellular hypha that had formed the haustorial mother cell, (iii) growth of the intracellular hypha within the newly invaded plant cell, (iv) formation by the intracellular hyphae of haustorial mother cells against the plant cell wall adjacent to an uninvaded cell, and (v) formation of haustoria in the uninvaded cell. These results support the previous suggestion that the haustorium may induce a state of susceptibility in the plant cell, "paving the way" for invasion by the intracellular hyphae.



1988 ◽  
Vol 66 (4) ◽  
pp. 736-741 ◽  
Author(s):  
Michèle C. Heath ◽  
C. J. Perumalla

The development of infection structures by the rust fungus Uromyces vignae was observed on oil-containing collodion membranes. About 40% of infection hyphae formed a haustorial mother cell, but this structure commonly senesced and died more rapidly than the infection hypha to which it was attached. These data suggest that the continued development of the haustorial mother cell requires some component normally provided by the host plant. Before they died, many haustorial mother cells apparently formed the thickened region of the wall which normally is traversed by the penetration peg during haustorium formation. Such a peg was observed in the centre of up to 40% of these thickened regions. However, no pegs protruded beyond the haustorial mother cell far enough to be called a haustorial neck. The thickened region of the haustorial mother cell wall could be differentiated from the rest of the wall by its lack of fluorescence under ultraviolet irradiation when mounted in Calcofluor or SITS (4-acetomido-4′-iso-thiocyanatostilbene-2,2′-disulphonic acid). Treatment with alkali, acid, chloroform–methanol, protease, and laminarinase did not affect this differential fluorescence, and the haustorial mother cell wall stained uniformly for proteins, carbohydrates, and chitin. Since Calcofluor normally binds to chitin, these data suggest that the thickened region of the haustorial mother cell wall may physically exclude the dye or may contain potential binding sites that are masked by other wall components.



2011 ◽  
Vol 57 (5) ◽  
pp. 382-391 ◽  
Author(s):  
Melissa J. Day ◽  
Randolph S. Currah

Three darkly pigmented species of conidial fungi of the family Pleosporaceae isolated from plants colonizing the Saskatchewan Glacier forefield were examined for potential roles in the degradation of moss gametophytes. Curvularia inaequalis and Ulocladium atrum isolated from bryophytes Ditrichum flexicaule and Tortella tortuosa , respectively, and Chalastospora gossypii from Saxifraga oppositifolia were inoculated onto autoclaved gametophytes of the moss Hylocomium splendens. All three species of fungi caused mass losses of the moss gametophytes. In vitro enzymatic tests revealed that all three fungi degraded cellulose, while none degraded insoluble polyphenols. When this material was examined by scanning electron microscopy, it was evident that the fungi had eroded the outer wall layer of the moss leaf cells to some extent but not the inner layer containing more lignin-like compounds. Once the outer wall layer was removed, the cells easily disarticulated. It is proposed that accumulations of these phenolics-rich leaf fragments subsequently ameliorate the rooting environment for vascular plants and have the potential to support the growth of basidiomycetes and other fungi, potentially mycorrhizal with pioneer vascular plants.



2009 ◽  
Vol 22 (3) ◽  
pp. 282-290 ◽  
Author(s):  
Minna Haapalainen ◽  
Kristin van Gestel ◽  
Minna Pirhonen ◽  
Suvi Taira

Type III protein secretion is essential for the pathogenicity of Pseudomonas syringae on its host plants. Expression of HrpA, a major component of the type III secretion system (T3SS)-associated pilus, was studied both in plant leaves and in vitro using reporter genes. We found that induction of the hrpA promoter was stronger in plants than in vitro, and that the induction was enhanced by both host and nonhost plants of P. syringae pv. tomato. In vitro, the expression was enhanced by cell-free exudates from plant cell suspension cultures, added into the minimal medium. Further analysis of the plant-cell-derived, hrpA-inducing factors showed that they were small and water-soluble compounds, which could signal P. syringae the proximity of living plant cells. We also studied the production and secretion of native HrpA protein in vitro, and detected a plant-signal-dependent increase in HrpA secretion. In contrast to HrpA, the intracellular accumulation or secretion of the other T3SS-dependent proteins were not significantly increased, despite the presence of plant cell-derived, promoter-inducing factors. Thus, the accumulation of HrpA pilin seems to be subjected to a distinct post-transcriptional regulation.



2004 ◽  
Vol 186 (14) ◽  
pp. 4685-4693 ◽  
Author(s):  
Kelly Poliquin ◽  
Yuri V. Ershov ◽  
Francis X. Cunningham ◽  
Tinsay T. Woreta ◽  
R. Raymond Gantt ◽  
...  

ABSTRACT In cyanobacteria many compounds, including chlorophylls, carotenoids, and hopanoids, are synthesized from the isoprenoid precursors isopentenyl diphosphate (IPP) and dimethylallyl diphosphate. Isoprenoid biosynthesis in extracts of the cyanobacterium Synechocystis strain PCC 6803 grown under photosynthetic conditions, stimulated by pentose phosphate cycle substrates, does not appear to require methylerythritol phosphate pathway intermediates. The sll1556 gene, distantly related to type 2 IPP isomerase genes, was disrupted by insertion of a Kanr cassette. The mutant was fully viable under photosynthetic conditions although impaired in the utilization of pentose phosphate cycle substrates. Compared to the parental strain the Δsll1556 mutant (i) is deficient in isoprenoid biosynthesis in vitro with substrates including glyceraldehyde-3-phosphate, fructose-6-phosphate, and glucose-6-phosphate; (ii) has smaller cells (diameter ca. 13% less); (iii) has fewer thylakoids (ca. 30% less); and (iv) has a more extensive fibrous outer wall layer. Isoprenoid biosynthesis is restored with pentose phosphate cycle substrates plus the recombinant Sll1556 protein in the Δsll1556 supernatant fraction. IPP isomerase activity could not be demonstrated for the purified Sll1556 protein under our in vitro conditions. The reduction of thylakoid area and the effect on outer wall layer components are consistent with an impairment of isoprenoid biosynthesis in the mutant, possibly via hopanoid biosynthesis. Our findings are consistent with an alternate metabolic shunt for biosynthesis of isoprenoids.



1993 ◽  
Vol 71 (9) ◽  
pp. 1236-1242 ◽  
Author(s):  
M. Stark-Urnau ◽  
K. Mendgen

Aecidiospores and uredospores are the two dikaryotic spore forms of the cowpea rust fungus Uromyces vignae. After germination they can be induced to develop a series of infection structures including appressoria, infection hyphae, and haustorial mother cells. Haustoria are then formed within host cells. The differentiation of infection structures was compared on polystyrene membranes with defined topographies, on scratched polyethylene membranes, and in planta. On polystyrene membranes with defined topographies both sporelings showed highest rates of differentiation on ridges 0.3 μm high but aecidiosporelings responded less efficiently to this stimulus than uredosporelings. On scratched polyethylene membranes, almost 90% of both sporelings differentiated appressoria, but only 10% formed haustorial mother cells; haustoria were not observed. On the host plant, by contrast, only 50% of the sporelings differentiated appressoria, but most of these formed haustorial mother cells and haustoria. In planta haustorial mother cell development occurred approximately 6 h earlier than on inductive membranes. Infection structures formed on artificial membranes and on host plants were similar in morphology and nuclear condition. Key words: cowpea rust fungus, nucleus, appressorium.



Author(s):  
Gunnel Karlsson ◽  
Jan-Olov Bovin ◽  
Michael Bosma

RuBisCO (D-ribulose-l,5-biphosphate carboxylase/oxygenase) is the most aboundant enzyme in the plant cell and it catalyses the key carboxylation reaction of photosynthetic carbon fixation, but also the competing oxygenase reaction of photorespiation. In vitro crystallized RuBisCO has been studied earlier but this investigation concerns in vivo existance of RuBisCO crystals in anthers and leaves ofsugarbeets. For the identification of in vivo protein crystals it is important to be able to determinethe unit cell of cytochemically identified crystals in the same image. In order to obtain the best combination of optimal contrast and resolution we have studied different staining and electron accelerating voltages. It is known that embedding and sectioning can cause deformation and obscure the unit cell parameters.



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