Cytochemical studies on Puccinia graminis f. sp. tritici in a compatible wheat host. I. Walls of intercellular hyphal cells and haustorium mother cells

1985 ◽  
Vol 63 (10) ◽  
pp. 1713-1724 ◽  
Author(s):  
J. Chong ◽  
D. E. Harder ◽  
R. Rohringer

Walls of intercellular hyphae and haustorium mother cells of the stem rust fungus in wheat leaves were studied cytochemically using lectin probes, periodate – thiocarbohydrazide – silver proteinate or periodate–chromate–phosphotungstate staining, and protease treatment. Up to six possible layers in the haustorium mother cell walls and four in the hyphal walls were resolved. Three outer layers of the haustorium mother cell walls were continuous with the three outer layers of the hyphal walls. The two innermost layers of the haustorium mother cell walls were not continuous with the hyphal walls but formed part of the septum. These two layers differed from the other layers of the haustorium mother cell walls in having no affinity to concanavalin A. In both hyphal and haustorium mother cell walls, components with an affinity for concanavalin A were extractable with protease treatment. Wheat-germ lectin binding occurred throughout the fungal walls except in the two outermost layers. Periodate-sensitive glycosubstances were also common, but the amounts varied among layers. Although some of these glycosubstances were confirmed as polysaccharides containing sugars with vicinal hydroxyl groups, much of the glycosubstances present in the walls was sensitive to protease treatment, indicating a possible glycoprotein composition.

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.


1986 ◽  
Vol 64 (11) ◽  
pp. 2561-2575 ◽  
Author(s):  
J. Chong ◽  
D. E. Harder ◽  
R. Rohringer

Various cytochemical tests on the wheat stem rust fungus were used to determine differences in components of the walls of the haustorium mother cell at the host cell penetration site and the haustorial neck and body and to describe some of the chemical properties of the extrahaustorial matrix. There were two transition zones with respect to wall composition. The first was at the host cell penetration site; chitin, present in haustorium mother cell walls, was not detected in haustorial neck walls. The second transition zone was at the neck ring; compared with walls of the proximal neck region, those distal to the neck ring contained more protein and lost much of their periodate – thiocarbohydrazide – silver proteinate reactive material and all concanavalin A binding material after treatment with protease. The two wall layers of the distal part of the haustorial neck were continuous with those of the haustorium; the wall layers of young haustorial bodies shared their staining properties and lectin affinities with those of the distal part of the haustorial necks, reflecting their common origin. As the haustoria matured, their body walls bound wheat germ lectin, but the neck walls did not. Tests indicated that polysaccharide and glycoprotein were present in the extrahaustorial matrix.


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.


1989 ◽  
Vol 67 (12) ◽  
pp. 3570-3579 ◽  
Author(s):  
C. W. Mims ◽  
J. Taylor ◽  
E. A. Richardson

Peanut rust disease proved to be an excellent system for ultrastructural study of development of infection structures by the fungus Puccinia arachidis. Fungal structures were clearly visible by light microscopy in fixed and embedded samples and could be located either on leaf surfaces or within the large substomatal chambers of peanut leaves. Samples could easily be oriented for thin sectioning. The infection process was a highly orchestrated process involving precisely timed events and highly specialized structures. Infection pegs developed from appressoria over stomata and entered the leaf by growing into the openings between guard cells. Once past the rim formed by the guard cell walls, the infection peg expanded to form a substomatal vesicle in which a synchronous mitotic division of the four nuclei occurred. A primary infection hypha then developed from the vesicle and grew into the mesophyll of the leaf until its tip or side contacted a host cell. A septum then delimited a binucleate or trinucleate terminal haustorial mother cell from the remainder of the infection hypha. The wall of the haustorial mother cell became closely appressed to that of the host cell. Following differentiation of the haustorial mother cell, a penetration peg arose from it and penetrated the host cell wall. The peg invaginated the host cell plasma membrane as it elongated and then expanded at its tip to form the haustorium body into which most of the contents of the haustorial mother cell moved. Meanwhile, the primary infection hypha formed secondary hyphae that gave rise to additional haustorial mother cells and haustoria. Key words: Puccinia arachidis, peanut rust, infection process, ultrastructure.


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.


1986 ◽  
Vol 64 (11) ◽  
pp. 2476-2486 ◽  
Author(s):  
Lesley A. Wood ◽  
Michèle C. Heath

Growth of the sunflower rust fungus (Puccinia helianthi Schw.) was compared by light microscopy in sunflower leaves, in untreated French bean leaves, in bean leaves given a preinoculation heat treatment, and on collodion membranes. Results suggested that fungal growth was slightly reduced and the formation of haustorial mother cells was inhibited in untreated bean leaves. Haustorial mother cells, when present, did not form haustoria and adjacent mesophyll cell walls usually were highly refractive. Preinoculation heat treatment reduced the incidence of refractive cell walls and increased that of haustorial mother cells and haustoria. Ultrastructurally, infection hyphae in unheated bean leaves appeared unusually vacuolate and often contained wall appositions where they touched the plant cells. Silicalike deposits were present in and on mesophyll cell walls at most infection sites. In heated plants, necrotic haustoria with small bodies were seen at the few sites that lacked silicalike deposits. At other sites, the fungus appeared to have stopped growing during the formation of the penetration peg or the haustorial neck, and such necks were encrusted with silicalike material. At most sites, penetration pegs were occluded resulting in the unusual situation in which the haustorial mother cell remained seemingly alive in spite of the necrosis of the haustorium.


1982 ◽  
Vol 60 (12) ◽  
pp. 2914-2922 ◽  
Author(s):  
D. J. Gray ◽  
H. V. Amerson ◽  
C. G. Van Dyke

Haustoria formed by the monokaryotic stage of Cronartium quercuum f. sp. fusiforme in Pinus taeda differed from those of the dikaryotic phase in Quercus rubra. Monokaryotic (M) haustorial walls were continuous with the walls of relatively undifferentiated haustorial mother cells. The septate M-haustorial neck and expanded M-haustorial body were separated from the invaginated host plasmalemma by a sheath which was continuous with the host cell wall. Collars encasing sheaths were infrequently observed. Dikaryotic (D) haustoria were morphologically similar to M-haustoria; however, they differed in several respects when examined with TEM. The D-haustorial mother cell wall was thickened at the penetration site but a reduction in the number of wall layers occurred between the thickened portion of the mother cell and the D-haustorium. A darkly staining neckband was present in the wall of the nonseptate D-haustorial neck but was lacking in the M-haustorium. An extensive sheath separated the invaginated host plasmalemma from the D-haustorial wall distally from the neckband. However, the sheath was separated from the D-haustorial wall and from the host cell wall by an invaginated host plasmalemma doubled in the haustorial neck region proximally from the neckband.


2001 ◽  
Vol 67 (8) ◽  
pp. 3413-3417 ◽  
Author(s):  
Xuan Peng ◽  
Jun Sun ◽  
Chris Michiels ◽  
Dirk Iserentant ◽  
Hubert Verachtert

ABSTRACT Pediococcus damnosus can coflocculate withSaccharomyces cerevisiae and cause beer acidification that may or may not be desired. Similar coflocculations occur with other yeasts except for Schizosaccharomyces pombe which has galactose-rich cell walls. We compared coflocculation rates ofS. pombe wild-type species TP4-1D, having a mannose-to-galactose ratio (Man:Gal) of 5 to 6 in the cell wall, with its glycosylation mutants gms1-1 (Man:Gal = 5:1) and gms1Δ (Man:Gal = 1:0). These mutants coflocculated at a much higher level (30 to 45%) than that of the wild type (5%). Coflocculation of the mutants was inhibited by exogenous mannose but not by galactose. The S. cerevisiae mnn2 mutant, with a mannan content similar to that ofgms1Δ, also showed high coflocculation (35%) and was sensitive to mannose inhibition. Coflocculation of P. damnosus and gms1Δ (or mnn2) also could be inhibited by gms1Δ mannan (with unbranched α-1,6-linked mannose residues), concanavalin A (mannose and glucose specific), or NPA lectin (specific for α-1,6-linked mannosyl units). Protease treatment of the bacterial cells completely abolished coflocculation. From these results we conclude that mannose residues on the cell surface of S. pombe serve as receptors for a P. damnosuslectin but that these receptors are shielded by galactose residues in wild-type strains. Such interactions are important in the production of Belgian acid types of beers in which mixed cultures are used to improve flavor.


2008 ◽  
Vol 133 (1) ◽  
pp. 107-116 ◽  
Author(s):  
Pablo Bolaños-Villegas ◽  
Shih-Wen Chin ◽  
Fure-Chyi Chen

The development of new cultivars in Doritaenopsis Guillaum. & Lami orchids is often hindered by factors such as low seed count in hybrids. Cytological study may offer the ability to develop new hybrids by revealing cultivars with good chromosome pairing and high pollen viability, which are somewhat difficult to obtain under current breeding programs. Cross pollination, pollen viability, and chromosomal behavior during meiosis were analyzed to reveal the relation between seed fertility and capsule set in Doritaenopsis hybrids. The number of mature capsules harvested and their relative seed content were used as indices of crossing availability. The results of meiosis were evaluated according to pollen viability detected by fluorescein diacetate and quantification of sporad types by acid fuchsin staining. Chromosome number and pairing at meiosis were observed in root tips or in samples of pollen mother cells. A positive relation was found among high seed set, high frequency of viable tetrads, high degree of chromosome pairing, and low frequency of chromosomal aberrations such as inversions and translocations. On the basis of these factors, three types of hybrids could be distinguished. In type one hybrids, chromosomes paired as bivalents, pollen mother cells divided into tetrads, and capsule setting occurred after pollination of pollen acceptors. In type two hybrids, chromosomes remained mainly as univalents that developed into micromeiocytes, pollen mother cell division was disrupted, and seed recovery was low after pollination. Type three hybrids showed chromosomes paired mostly as multivalents, chromosome bridges, pollen mother cell division with massive failure, and little fertility. In Doritaenopsis orchids, high pollen viability and high fertility depends on a high frequency of normal tetrads, and low seed set in cross-pollination is predicted with micronuclei in the end products of meiosis. The occurrence of chromosomal aberrations may suggest a process of genome differentiation that could compromise breeding efforts if not taken into consideration.


1988 ◽  
Vol 256 (2) ◽  
pp. 661-664 ◽  
Author(s):  
M S Stoll ◽  
T Mizuochi ◽  
R A Childs ◽  
T Feizi

Conditions have been established for the rapid and efficient conjugation of reducing oligosaccharides (di- to deca-saccharides) to dipalmitoyl phosphatidylethanolamine. The resulting neoglycolipids derived from several naturally occurring oligosaccharides and a series of N-linked high-mannose-type oligosaccharides released by hydrazinolysis from RNAase B showed specific and potent reactivities, as appropriate, with monoclonal antibodies to blood group Lewis(b), blood group A or a stage-specific embryonic (SSEA-1) antigen, or the lectin concanavalin A.


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