• Mycobiology
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• v.33 no.1
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• pp.1-6
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• 2005

Morphological characteristics of hyphal interaction between Grifola umbellata (Pers. Ex Fr.) Pilat and its companion fungus which related to sclerotia formation from hyphae were investigated by external observations, light microscopy and transmission electron microscopy (TEM). External observations showed that a dense antagonism line was formed by both G. umbellata and companion fungus after their hyphae contacted each other in dual culture. Many hyphal strands emerged on the colony of G. umbellata and differentiated to sclerotia from where hyphal strands crossed. Light microscope observations revealed the process of antagonism line formation. Mature antagonism with structural differentiation, was composed of three main layers: the rind, the rind underlayer and the hypha layer. TEM observations showed that after colonies hyphal contact, a series of reactions always occurred in both G. umbellata and companion fungus. Cells in the center of antagonism line were dead. Cells of G. umbellata adjacent to the antagonism line were usually large and hollow, with unilateral thickened wall, whereas those of companion fungus were empty, with thin or thick wall. Both hyphal interaction at the antagonism line may be one of the main reasons for sclerotia of G. umbellata differentiation from hypha.

• ALGAE
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• v.20 no.4
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• pp.353-361
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• 2005

Ascophyllum nodosum (L.) Le Jolis has a systemic infection with the ascomycete Mycophycias ascophylli (Cotton) Kohlmeyer and Volkmann-Kohlmeyer with which it establishese a mutualistic symbiosis. In addition, A. nodosum is the host for the obligate red algal epiphyte, Vertebrata lanosa (L.) Christensen. Using light and electron microscopy we describe morphological and cytochemical changes occurring as a consequence of rhizoid penetration of V. lanosa into cortical host tissue. Rhizoids induce localized cell necrosis based on physical damage during rhizoid penetration. Host cells adjacent to the rhizoid selectively undergo a hypersensitive reaction in which they become darkly pigmented and become foci for hyphal development. Light and electron microscopy show that M. ascophylli forms dense hyphal aggregations on the surface of the V. lanosa rhizoid and extensive endophytic hyphal growths in the rhizoid wall. This is the first morphological evidence of an interaction between M. ascophylli and V. lanosa. We speculate that M. ascophylli may be interacting with V. lanosa to limit tissue damage to their shared host. In addition, the fungus provides a potential pathway for the transfer of materials (e.g., nutrients and photosynthate) between the two phototrophs.

• Korean Journal of Environmental Agriculture
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• v.23 no.4
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• pp.191-196
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• 2004

The contribution of plant nutrition status in arbuscular mycorrhizal (AM) plant to the nutrient acquisition by extraradical hyphae of AM fungi was investigated using cucumber colonized with Glomus intraradicies (BEG 110) focusing on the Zn. Compartmentalized pots with separated Bones for hyphal growth were used to determine the contribution of extraradical AM hyphae to Zn uptake from hyphal zones. $0.5\;{\mu}M$ Zn was supplied into the hyphal zones as nutrient solution (10 mL/day) with a form of $ZnSO_4$. Zn foliar application was made two times for one week before harvest (8 mL/plant). The colonization rate by AM were high in all of Zn treatments. The dry weight of cucumber increased by AM colonization compared to those of non-mycorrhizal counterpart. However: Zn foliar application resulted in no significant difference in dry weight between mycorrhizal- and non-mycorrhizal plant. In addition, the enhancement of Zn content in cucumber shoot by AM colonization were also reduced by Zn foliar application. These results indicate that the interaction between host plant and AM fungus for nutrient uptake might be related to plant nutritional status and nutrient contents. In consequence, higher Zn contents in host plant by foliar application of Zn could restrict the role of extraradical hyphae of AM fungus on the Zn acquisition and transfer from fungus to host plant.

• Journal of Microbiology and Biotechnology
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• v.31 no.6
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• pp.815-822
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• 2021

Indigenous fungus-feeding nematodes may adversely affect the growth and activity of introduced biocontrol fungi. Alginate pellets of the biocontrol fungus Trichoderma harzianum ThzID1-M3 and sclerotia of the fungal plant pathogen Sclerotinia sclerotiorum were added to nonsterile soil at a soil water potential of -50 or -1,000 kPa. The biomass of ThzID1-M3, nematode populations, and extent of colonization of sclerotia by ThzID1-M3 were monitored over time. The presence of ThzID1-M3 increased the nematode population under both moisture regimes (p < 0.05), and fungivores comprised 69-75% of the nematode population. By day 5, the biomass of ThzID1-M3b and its colonization of sclerotia increased and were strongly correlated (R² = 0.98), followed by a rapid reduction, under both regimes. At -50 kPa (the wetter of the two environments), fungal biomass and colonization by ThzID1-M3 were less, in the period from 5 to 20 days, while fungivores were more abundant. These results indicate that ThzID1-M3 stimulated the population growth of fungivorous nematodes, which in turn, reduced the biocontrol ability of the fungus to mycoparasitize sclerotia. However, colonization incidence reached 100% by day 5 and remained so for the experimental period under both regimes, although hyphal fragments disappeared by day 20. Our results suggest that indigenous fungivores are an important constraint for the biocontrol activity of introduced fungi, and sclerotia can provide spatial refuge for biocontrol fungi from the feeding activity of fungivorous nematodes.

• ALGAE
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• v.20 no.3
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• pp.225-232
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• 2005

Optical microscopy of recently living and cleared material of the fucoid, Ascophyllum nodosum (L.) Le Jolis, revealed novel aspects of its interaction with the ascomycete Mycophycias ascophylli (Cotton) Kohlmeyer and Kohlmeyer (previously Mycosphaerella ascophylli Cotton). Most host cells are associated with hyphae by lateral attachment of cell walls. Hyphae form extensive networks throughout the host thallus and show considerable differentiation in the various host tissues. In the base of epidermal cells, hyphae form multicellular rings around each host cell to produce a continuous network. In medullary regions, long, relatively unbranched and longitudinally aligned hyphae occur, with radial branches extending into cortical regions. Scattered in the inner cortex of host tissue are numerous multicellular nodes of smaller, polygonal to irregular shaped cells with five or more radiating arms of hyphae. Individual hyphal cells show a variety of specializations including swellings and appressoria-like attachments to some host cells. These observations provide the morphological basis for the mutualistic symbiosis supported by recent experimental work. We conclude that this association is best described by the term “symbiotum.”

• Journal of Microbiology
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• v.44 no.3
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• pp.311-319
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• 2006

Bud6p is a component of a polarisome that controls cell polarity in Saccharomyces cerevisiae. In this study, we investigated the role of the Candide albicans Bud6 protein (CaBud6p) in cell polarity and hyphal development. CaBud6p, which consists of 703 amino acids, had 37% amino-acid sequence identity with the Bud6 protein of S. cerevisiae. The homozygous knock-out of CaBUD6 resulted in several abnormal phenotypes, such as a round and enlarged cells, widened bud necks, and a random budding pattern. In hypha-inducing media, the mutant cells had markedly swollen tips and a reduced ability to switch from yeast to hypha. In addition, a yeast two-Hybrid analysis showed a physical interaction between CaBud6p and CaAct1p, which suggests that CaBud6p may be involved in actin cable organization, like Bud6p in S. cerevisiae. Taken together, these results indicate that CaBud6 plays an important role in the polarized growth of C. albicans.

• The Korean Journal of Mycology
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• v.36 no.2
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• pp.153-162
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• 2008

Pinewood nematode (PWN) trapping by nematophagous fungi, Arthrobotrys conoides, A. dactyloides and A. oligospora and the fungal growth were characterized. The three Arthrobotrys species each was inoculated into the PWN cultured on Botrytis cinera fungal colony on potato dextrose agar (PDA). The effects of temperature, pH, PWN inoculation density and nutrients on the growth of the three Arthrobotrys spp were measured. A. conoides grew fast, 13.9 mm/day while A. dactyloides grew slow, 3 mm/day. PDA medium was the best for the fungal growth at $25^{\circ}C$ and pH 4.5. The Arthrobotrys spp growth was stimulated by 500 nematodes inoculation but not by 1000 inoculation. A. dactyloides did not grow below pH 4.5 and at high PWN density. A. conoides and A oligospora formed trapping organs with thick constricting hyphal network only when PWN present, while A. dactyloides formed the organ with circular hyphae constitutively. A. conoides formed trapping organs faster than A. oligospora did. The nematode trapping hyphae of the fungi penetrated into PNW inside to form many tiny infection bulbs and to digest the nematode. However, A. dactyloides formed a few trapping organs but no trapping was observed. Infection rate of PWN was 95% by A. conoides, 80% by A. oligospora and 92% by the combination inoculation of A. conoides and A. oligospora. In contrast A. dactyloides increased PWN density without infecton. There was no interaction effect in any combination inoculation of the three Arthrobotrys spp. A. conoides enhanced PWN infection rate by rapid hyphal growth and early trapping, while A. oligospora did it by increasing hyphal density. In conclusion A. conoides is the most effective in both hyphal growth and infection, and thus these characteristics can be utilized as a biological control of PWN.

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.831-838
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• 2018

Trophic interactions of introduced biocontrol fungi with soil animals can be a key determinant in the fungal proliferation and activity. This study investigated the trophic interaction of an introduced biocontrol fungus with soil nematodes. The biocontrol fungus Trichoderma harzianum ThzID1-M3 and the fungivorous nematode Aphelenchoides sp. (10 per gram of soil) were added to nonsterile soil, and microbial populations were monitored for 40 days. Similar results were obtained when the experiment was duplicated. ThzID1-M3 stimulated the population growth of indigenous nematodes (p < 0.05), regardless of whether Aphelenchoides sp. was added. Without ThzID1-M3, indigenous nematodes did not increase in number and the added Aphelenchoides sp. nematodes almost disappeared by day 10. With ThzID1-M3, population growth of nematodes was rapid between 5 and 10 days after treatment. ThzID1-M3 biomass peaked on day 5, dropped at day 10, and then almost disappeared at day 20, which was not influenced by the addition of nematodes. In contrast, a large quantity of ThzID1-M3 hyphae were present in a heat-treated soil in which nematodes were eliminated. Total fungal biomass in all treatments peaked on day 5 and subsequently decreased. Addition of nematodes increased the total fungal biomass (p < 0.05), but ThzID1-M3 addition did not affect the fungal biomass. Hyphae of total fungi when homogenously distributed did not support the nematode population growth; however, hyphae of the introduced fungus did when densely localized. The results suggest that soil fungivorous nematodes are an important constraint on the hyphal proliferation of fungal agents introduced into natural soils.

• The Plant Pathology Journal
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• v.21 no.4
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• pp.301-309
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• 2005

Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from $11\;to\;57\%$. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc ($23-57\%$) and Hy ($16-47\%$). Different enzymes (trypsin, pepsin, proteinase k, a-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin, cycloheximide, brefeldin A, sodium azide, dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments. Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv-25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv-25/98, respectively. Trypsin, chymotrypsin and b-1,3-glucanase totally inhibited lectin activity. Similarly, various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectin-carbohydrate interaction.

• Mycobiology
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• v.46 no.4
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• pp.407-415
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• 2018

Pheromone (PHB)-receptor (RCB) interaction in the mating pheromone response pathway of Lentinula edodes was investigated using synthetic PHBs. Functionality of the C-terminally carboxymethylated synthetic PHBs was demonstrated by concentration-dependent induction of a mating-related gene (znf2) expression and by pseudoclamp formation in a monokaryotic strain S1-11 of L. edodes. Treatment with synthetic PHBs activated the expression of homeodomain genes (HDs) residing in the A mating type locus, and of A-regulated genes, including znf2, clp1, and priA, as well as genes in the B mating type locus, including pheromone (phb) and receptor (rcb) genes. The synthetic PHBs failed to discriminate self from non-self RCBs. PHBs of the B4 mating type (B4 PHBs) were able to activate the mating pheromone response pathway in both monokaryotic S1-11 and S1-13 strains, whose B mating types were B4 (self) and B12 (non-self), respectively. The same was true for B12 PHBs in the B4 (non-self) and B12 (self) mating types. The synthetic PHBs also promoted the mating of two monokaryotic strains carrying B4-common incompatible mating types ($A5B4{\times}A1B4$). However, the dikaryon generated by this process exhibited abnormally high content of hyphal branching and frequent clamp connections and, more importantly, was found to be genetically unstable due to overexpression of mating-related genes such as clp1. Although synthetic PHBs were unable to discriminate self from non-self RCBs, they showed a higher affinity for non-self RCBs, through which the mating pheromone response pathway in non-self cells may be preferentially activated.