不同物质对白鬼笔菌丝生长的影响

本文研究了不同物质对白鬼笔菌丝生长的影响.采用平板培养法,以白鬼笔菌株Phallus impudicus-1为供试菌株,分析了 5种物质对白鬼笔菌丝生长的影响.供试碳源中,当以红糖为碳源时,白鬼笔菌丝生长速率最快,为1.445mm/d,蔗糖和葡萄糖次之;供试氮源中,当以酵母膏为氮源时,白鬼笔菌丝浓密,菌丝生长速率最快,...     

Translocation of soil-derived phosphorus in mycelial cord systems in relation to inoculum resource size

Abstract: Uptake of ³²P phosphorus from soil was investigated in mycelial cord systems of Phanerochaete velutina, Hypholoma fasciculare, Tricholomopsis platyphylla and Phallus impudicus which extended from 0.5, 2, 4 or 8 cm³ beech ( Fagus sylvatica ) inocula. Cord systems accumulated between 4.8 and 18.7% of phosphorus supplied to soil, according to species and size of inoculum. Phosphorus translocation to newly-colonized 2 cm³ beech baits, determined non-destructively, was characterized by an initial steady phase, of 2.5 to 32 nmol P day⁻¹ which lasted at least 12 days for all four species. After the initial steady phase, translocation rates declined. Initial mycelial extension and wood decay rates also varied with species and inoculum size. There was no clear relationship between phosphorus translocation rates, wood decay or the distribution of soil-derived phosphorus in cord system components. However, with increasing inoculum size, P. velutina systems allocated a significantly greater proportion of available phosphorus to newly-colonized baits. The degree to which distribution of soil-derived phosphorus in cord systems is related to nutrient conservation or metabolic demand in the fungi is discussed.     

Translocation of 32P between wood resources recently colonised by mycelial cord systems of Phanerochaete velutina

Abstract The basidiomycete fungus Phanerochaete velutina was inoculated centrally into trays of compressed, non-sterile woodland soil on precolonised 1 cm³ beech wood blocks. Mycelial systems developed from this and colonised two ‘baits’ (wood blocks or inert plastic controls), one on either side of the inoculum block. ³²P-orthophosphate was supplied to a bait and its appearance in the other bait was monitored non-destructively with time, and destructively by liquid scintillation counting 60 or 70 days after addition of the radioisotope. Phosphorus was taken up by the first bait, translocated back to the inoculum block and onwards to the second bait. When the second bait was added 10 days after the first, translocation to the former was much more rapid indicating a large demand for phosphorus during early stages of colonisation. The size of the bait to which the ³²P was added appeared to determine the amount taken up by the whole system, and the size of the second bait determined how much ³²P was translocated to it. Decayed and/or small baits had less demand for phosphorous. The ecological relevance of these findings is discussed.     

Growth morphology of Streptomyces akiyoshiensis in submerged culture: influence of pH, inoculum, and nutrients.

Most media in which the growth of shaken submerged cultures of Streptomyces akiyoshiensis was examined did not support the formation of well-dispersed mycelial suspensions. Investigation of the culture conditions promoting dispersed growth showed the pH of the culture medium to be of critical importance; an initial value of 5.5 minimized aggregation of the mycelium while supporting adequate biomass production. In cultures started at this pH, spore inocula gave better mycelial dispersal than did vegetative inocula; with spore inocula, growth morphology was also less affected by inoculum size. The composition of the nutrient solution influenced the extent of mycelial dispersal; slow growth was often associated with clumping but no clear correlation was observed between pellet formation and the ability of carbon or nitrogen sources to support rapid growth. Increasing the phosphate concentration from 0.5 to 15 mM caused a modest decrease in mycelial aggregation. Conditions promoting a well-dispersed mycelium suitable for studying the physiological control of secondary metabolism also supported the formation of 5-hydroxy-4-oxonorvaline by S. akiyoshiensis.     

Microcycle Conidiation and Spore-Carrying Capacity of Colletotrichum gloeosporioides on Solid Media

The effect of spore inoculum density, medium concentration, and temperature on slime-spot formation, spore yield, and mycelium production by Colletotrichum gloeosporioides on agar media were studied with a simple microplate assay. A steady-state spore yield (spore-carrying capacity) independent of inoculum density was reached only on media that supported good fungal growth and sporulation. The spore-carrying capacity was reached earlier, the denser the inoculum. On standard mycological media a high inoculum density (2.5 × 10⁶ spores per ml) resulted in a slimy mass of conidia forming a slime spot, a phenomenon associated with greatly reduced mycelium formation and indicative of microcycle conidiation. In contrast, for a similar inoculum density, enhanced mycelial growth preceded sporulation and overrode slime-spot formation on highly concentrated media; a very low medium concentration resulted in much less mycelium, but spore production was also decreased. Exposure to suboptimal growth temperatures of 36 to 48°C for up to 8 days did not induce microcycle conidiation from inocula that did not form a slime spot at 28°C.     

A Unique Pattern of Mycelial Elongation of Blakeslea trispora and Its Effect on Morphological Characteristics and β-Carotene Synthesis

The mycelial morphology of Blakeslea trispora was of crucial importance in the production of beta-carotene in submerged cultures of B. trispora. After the spores were inoculated, the time-course variation of mycelial morphology was closely examined under the microscope. With the addition of the non-ionic surfactant (Span 20: Sorbitan monolaurate, E493) to the culture medium, a unique pattern of mycelial elongation was observed: 1) slow formation of germ tubes from spores and 2) appearance of mycelia with very short length, which allowed a well-dispersed growth of B. trispora without significant pellet aggregation. Span 20 appears to act like a paramorphogen. Without Span 20, however, the fungal culture finally formed a big clump of mycelium owing to heavy cross-linking of long mycelia. But the short mycelium maintained in the course of cultivation seemed to be irrelevant to growth inhibition, because the final concentration of dry mycelium was much higher with Span 20 after 3-day cultivation. The 20-fold increase in specific yield of beta-carotene (mg beta-carotene produced per g mycelium) was achieved with this drastic change in the pattern of mycelial elongation. The reason for this result might be more effective mass transfer and/or enhanced sensitivity to environmental oxidative stress in the well-dispersed mycelial cultures of B. trispora.     

The induction of mycelial development inAureobasidium pullulans (IMI 45533) by yeast extract

Germ tube and subsequent mycelial development from yeast-like and swollen cells ofAureobasidium pullulans (IMI 45533) was induced by yeast extract in defined liquid medium. This morphogenetic transition was dependent on inoculum size; pH effects were not involved and once mycelial development was induced in the cells it continued even in the absence of yeast extract. The progeny of mycelium and future generations were unaffected by yeast extract. Cessation of germination was not due to any obvious medium changes but appeared to be partly due to the production of a germination inhibitor, which could also be produced by control cells grown in the absence of yeast extract.     

Effect of culture conditions on ergosterol as an indicator of biomass in the aquatic hyphomycetes

Ergosterol is a membrane component specific to fungi that can be used to estimate fungal biomass using appropriate factors of conversion. Our objectives were to determine the limits of use of ergosterol content as a measure of biomass for aquatic hyphomycetes, and to evaluate a previously established ergosterol-to-biomass conversion factor. We varied inoculum quality, growth medium, and degree of shaking of four aquatic hyphomycete species. In cultures inoculated with homogenized mycelium, we found a significant effect of shaking condition and culture age on ergosterol content. In liquid cultures with defined medium, ergosterol content reached 10 to 11 microg/mg of mycelium (dry mass) and varied by factors of 2.2 during exponential growth and 1.3 during stationary phase. The increase in ergosterol content during exponential phase could be attributed, at least in part, to rapid depletion of glucose. Oxygen availability to internal hyphae within the mycelial mass is also responsible for the differences found between culture conditions. Ergosterol concentration ranged from 0.8 to 1.6 microg/mg in static cultures inoculated with agar plugs. Ergosterol content varied by a factor of 4 in two media of different richnesses. For different combinations of these parameters, strong (r(2) = 0.83 to 0.98) and highly significant (P < 0.001) linear relationships between ergosterol and mycelial dry mass (up to 110 mg) were observed. Overall, the ergosterol content varied by a factor of 14 (0.8 to 11 mg/g). These results suggest that care must be taken when the ergosterol content is used to compare data generated in different field environments.     

Glutamine requirement for aerial mycelium growth in Neurospora crassa

Five amino acids are accumulated during vegetative growth of Neurospora crassa, particularly.during the prestationary growth phase. Alanine, glutamine, glutamate, arginine and ornithine.comprised over 80% of the total amino acid pool in the mycelium. Amino acid pools of different amino acid auxotrophs were followed during the partial transformation of a mycelial mat into an aerial mycelium. The mycelial mat under starvation and in direct contact with air rapidly formed aerial mycelium, which produced thereafter a burst of conidia. During this process,glutamine and alanine in the mycelial mat were consumed more rapidly than other amino acids;in the growing aerial mycelium, glutamate and glutamine were particularly accumulated. Of the amino acids that were initially accumulated in the mycelial mat, only a high glutamine pool was required for aerial mycelium growth induced by starvation. This requirement for glutamine could not be satisfied by a mixture of the amino compounds that are synthesized via glutamine amidotransferase reactions. It is proposed that glutamine serves as a nitrogen carrier from the mycelial mat to the growing aerial mycelium.     

Antibiotic Production of Hyphal Fractions of Streptomyces griseus: II. Streptomycin Production of Different Fractions Obtained by Density Gradient Centrifugation

The streptomycin-producing activity (SPA) of hyphal fractions from washed mycelium of submerged cultures of Streptomyces griseus strain 52-1, as obtained by density gradient centrifugation, was investigated. Activity of the various fractions differed strongly in intensity. The highest SPA was evident in the unfractionated mycelium. A synergistic effect upon SPA was found in the interaction of cultures of different ages, and a 55% increase in yield was obtained by mixing the 48- and 72-hr cultures. A synergistic effect occurred in all combinations studied. By use of fractions obtained from 72-hr mycelium for inoculation, differences in streptomycin production were noted. Some inoculum fractions yielded a greater amount of streptomycin (36%) than the unfractionated mycelial inoculum.     

Production of acid proteinases byHumicola lutea immobilized in polyacrylamide gel

Summary Fungal spores ofHumicola lutea 120–5 were entrapped in 5% polyacrylamide gel and were cultivated for 44–48 h to form a mycelial network inside the beads. A dense mycelial growth also occurred on the surface of the beads. It was possible to reuse the immobilized mycelium for production of acid proteinases in 12 different batches without loss of mechanical stability. The inoculum size should be controlled prior to its transfer into fresh production medium. Maximal enzyme production exceeding the level of free cell fermentation was registered in the fourth to seventh cycles. According to the size of the inoculum, half of the initial production rate was reached after 7–14 batches.     

A Unique Pattern of Mycelial Elongation of Blakeslea trispora and Its Effect on Morphological Characteristics and β-Carotene Synthesis

The mycelial morphology of Blakeslea trispora was of crucial importance in the production of β-carotene in submerged cultures of B. trispora. After the spores were inoculated, the time-course variation of mycelial morphology was closely examined under the microscope. With the addition of the non-ionic surfactant (Span 20: Sorbitan monolaurate, E493) to the culture medium, a unique pattern of mycelial elongation was observed: 1) slow formation of germ tubes from spores and 2) appearance of mycelia with very short length, which allowed a well-dispersed growth of B. trispora without significant pellet aggregation. Span 20 appears to act like a paramorphogen. Without Span 20, however, the fungal culture finally formed a big clump of mycelium owing to heavy cross-linking of long mycelia. But the short mycelium maintained in the course of cultivation seemed to be irrelevant to growth inhibition, because the final concentration of dry mycelium was much higher with Span 20 after 3-day cultivation. The 20-fold increase in specific yield of β-carotene (mg β-carotene produced per g mycelium) was achieved with this drastic change in the pattern of mycelial elongation. The reason for this result might be more effective mass transfer and/or enhanced sensitivity to environmental oxidative stress in the well-dispersed mycelial cultures of B. trispora.     

Glutamine metabolism during aerial mycelium growth of Neurospora crassa

During vegetative growth, glutamine is accumulated in the mycelium of Neurospora crassa. This high pool of glutamine seems to be required for aerial mycelium growth. Enzymes responsible for the synthesis and catabolism of glutamine were measured before and during the partial transformation of a mycelial mat into aerial mycelium. In the transforming mycelial mat,considerable activities of the biosynthetic NADP-glutamate dehydrogenase and glutamine synthetase (predominantly β polypeptide) and also some activity of glutamate synthase were observed. In the aerial mycelium, glutamine synthetase (predominantly β polypeptide) was detected, but very low activities of NADP-glutamate dehydrogenase and glutamate mycelium could derive from glutamine. No glutaminase activity could be detected. It is suggested that glutamate is formed through the activities of the glutamine transaminase-ω -amidase pathway and another transaminase. High activities of glutamine and alanine transaminases were observed in the aerial mycelium. These results are discussed in terms of the possible role of glutamine as a nitrogen carrier from the mycelium to the growing aerial hyphae.     

Investigations on the causes of upper stem rot (USR) on standing mature oil palms

Three different trials to examine the cause of upper stem rot (USR) infection in oil palm failed to achieve any infection. In the first experiment, inoculum was applied as colonised rubber wood blocks or as spore suspensions. In the second experiment, particular attention was given to ensure that the Ganoderma spores were freshly collected to maintain viability but no infection was observed around the inoculation sites of any of the different oil palm tissues treated. Lastly in the third experiment, both monokaryotic and dikaryotic mycelial cultures were applied directly to cut fronds, which were protected with a moist covering, but no infection was detected after more than two years. Failure to achieve infection by direct inoculation would indicate that USR does not arise from direct infection of living tissues by Ganoderma spores or mycelium, this is probably because of insufficient inoculum potential to cause infection. It is suggested that USR infection is achieved only when a sufficiently large source of inoculum has built up in dead material, probably in frond axils, and this allows invasion of the living tissues.     

Growth induced translocation effectively directs an amino acid analogue to developing zones in Agaricus bisporus

The vegetative mycelium of Agaricus bisporus supplies developing white button mushrooms with water and nutrients. However, it is not yet known which part of the mycelium contributes to the feeding of the mushrooms and how this depends on growth conditions. Here we used photon counting scintillation imaging to track translocation of the ¹⁴C-radiolabeled metabolically inert amino acid analogue α-aminoisobutyric acid (¹⁴C-AIB). Translocation to the periphery of the mycelium was observed in actively growing vegetative mycelium with a velocity of up to 6.6 mm h⁻¹, which was 30-fold higher than the growth rate. Furthermore, ¹⁴C-AIB translocated to neighboring colonies after fusion by anastomosis depending on the relative growth rate in these colonies. When mushrooms started to develop, translocation of ¹⁴C-AIB was redirected to the fruiting bodies via mycelium and hyphal cords. More abundant mycelial cord formation and a 5-fold higher rate of translocation was observed for cultures growing directionally from inoculum located at one side of the substrate, when compared to non-directional growth (inoculum mixed throughout the substrate). The maximum translocation distance was also greater (≥50 and 22 cm, respectively). In conclusion, ¹⁴C-AIB translocation switches between vegetative growth and towards developing mushrooms, especially via cords and when source–sink relationships change.     

Influence of culture media and environmental factors on mycelial growth and pycnidial production of Sphaeropsis pyriputrescens

Sphaeropsis pyriputrescens, the causal agent of Sphaeropsis rot of pears and apples, is a recently described species. In this study the effects of culture media, temperature, water potential, pH and light on mycelial growth and pycnidial production of S. pyriputrescens were evaluated. Apple juice agar and pear juice agar were most suitable for mycelial growth of all six isolates tested. Cornmeal agar was not suitable for either mycelial growth or pycnidial production. The fungus grew from -3 to 25 C, with optimum growth at 20 C and no growth at 30 C. The fungus grew at water potential as low as -5.6 MPa on potassium chloride-amended potato-dextrose agar (PDA). Hyphal extension was not observed at -7.3 MPa after 10 d incubation, but growth resumed when the inoculum plugs were placed on PDA. The fungus grew at pH 3.3-6.3 and optimum growth was at pH 3.3-4.2. No mycelial growth was observed at pH above 7.2 after 10 d incubation, but growth resumed when the inoculum plugs were transferred onto PDA. Regardless of medium tested, few pycnidia formed at 20 C in the dark. Pycnidial production was enhanced significantly by fluorescent light, but continuous light appeared to reduce pycnidial production, depending on the medium. Oatmeal agar (OMA) was most suitable for production of pycnidia and conidia. Pycnidia that formed on 3 wk old OMA cultures at 20 C under 12 h light/12 h dark produced abundant conidia, and the technique is recommended for inoculum production.     

The reactions of susceptible and resistant tomato cultivars to strains of Verticillium albo-atrum

Three strains of Verticillium albo-atrum causing severe wilt of tomato (T), progressive (Hp ) and fluctuating (HF) wilt of hop, were inoculated through the roots of four tomato cultivars at different inoculum concentrations. Symptoms were assessed visually 42 days after inoculation, and quantitatively on the change in total leaf area compared with controls. Distribution of mycelium and tyloses was determined by sections at 2 cm intervals of root, stem and petiole. Cultivars Loran Blood and Moscow showed resistance to disease expression at all levels of inoculum concentration with the T strain. Bonny Best and Potentate were both susceptible to this strain, but whereas in Potentate, disease severity increased from mild to severe with increase in inoculum concentration, Bonny Best was severely diseased at the lowest level of inoculum. All cultivars showed some susceptibility to the HP and HF strains; the ‘resistance’ of Loran Blood and Moscow was no longer apparent and Bonny Best was most severely affected. The relative susceptibilities to the strains were HF Bonney Best > Loran Blood > Potentate > Moscow, HP Bonny Best > Loran Blood, Moscow > Potentate, T Bonny Best > Potentate > Loran Blood, Moscow. In general, vascular colonization was less in the cultivars Loran Blood and Moscow with all three fungal strains at io⁵propagules/ml level of inoculum, but this was not always correlated with an increase in disease severity. With the exception of the host-pathogen combinations Bonny Best/T, Bonny Best/HF, Potentate/T and Moscow/T, increasing the inoculum concentration to 10⁷propagules/ml increased disease severity but had little or no effect of increasing vascular colonization. In Bonny Best/T, Bonny Best/HF and Potentate/T vascular colonization was reduced with the higher level of inoculum. Moscow showed complete resistance to symptom expression and little vascular colonization with the T strain at 10⁵prop./ml. At 10⁷prop./ml resistance was maintained but there was very extensive growth of mycelium in the vessels. Tylosis resulted from an interaction of host, fungal strain and the level of inoculum and was not always correlated with the degree of vascular colonization. Contrary to previous reports the resistant varieties Loran Blood and Moscow developed acute disease symptoms after inoculation with HP and HF and these were associated with a high level of tylosis rather than mycelial growth. Tylosis and disease severity but not mycelial growth increased with higher levels of inoculum. The results suggested that susceptibility to Verticillium wilt was a complex response depending on host cultivar, fungal strain and the initial inoculum concentration. In some cultivar-pathogen combinations susceptibility was directly proportional to the amount of mycelium present in the vessels, while in others a physiological resistance mechanism independent of the degree of colonization appeared to operate. In a third category, increased disease development rather than resistance was associated with high levels of tylosis.     

Production of thermostable and neutral glucoamylase using immobilized <Emphasis Type="Italic">Thermomucor indicae-seudaticae</Emphasis>

Thermomucor indicae-seudaticae was immobilized in alginate, κ-carrageenan, agarose, agar, polyacrylamide and loofah (Luffa cylindrica) sponge (as such or coated with alginate/starch/Emerson YpSs agar), and used for the production of glucoamylase in submerged fermentation. The mycelium developed from alginate-immobilized sporangiospores secreted higher glucoamylase titres (22.7 U ml⁻¹) than those immobilized in other gel matrices and the freely growing mycelial pellets (18.5 U ml⁻¹). Loofah network provided a good support for mycelial growth, but the enzyme production was lower than that attained with alginate beads. Glucoamylase production increased with inoculum density and the optimum levels were achieved when 40 calcium alginate beads (∼5 × 10⁶ immobilized spores) were used to inoculate 50 ml production medium. The alginate bead inoculum displayed high storage stability at 4°C and produced comparable enzyme titres up to 120 days. The glucoamylase production by hyphae emerged from the immobilized sporangiospores was almost stable over eight batches of repeated fermentation. Scanning electron micrographs of alginate beads, after batch fermentation, revealed extensive mycelial growth inside and around the beads.     

Grazing alters network architecture during interspecific mycelial interactions

The changes that occur in mycelial architecture of Phanerochaete velutina interacting with Hypholoma fasciculare mycelium in soil microcosms in the presence and absence of the collembola Folsomia candida are investigated employing tools developed in graph theory and statistical mechanics. There was substantially greater overgrowth of H. fasciculare by P. velutina mycelium when grazed than when un-grazed. There was a marked disappearance of hyphal links in all un-grazed systems between 8 d and 34 d, predominantly in areas distant from the interaction, but this was much less evident in grazed systems. Further, new tangential cross-links connecting radial cords distant from the inoculum formed in grazed systems. The thickness of cords increased with time, and more so in grazed systems. There was no significant difference in transport efficiency between the grazed and un-grazed systems. The ability of the mycelial network to modify dynamically link strengths is crucial to achieving a balance between transport capacity/robustness to damage and overall cost of production.     

Mechanisms for the development of genetically variable mycorrhizal mycelia in the ectomycorrhizal fungus Laccaria bicolor.

An in vitro study investigated mechanisms for the development of genetically variable mycorrhizal mycelia for Laccaria bicolor. Seedlings of jack pine (Pinus banksiana) grown nonaseptically in an autoclaved soil substrate were given different L. bicolor inoculum treatments. These included (i) a dikaryotic mycelium genotype (D); (ii) D and basidiospores collected from one group of five sporophores (T1); (iii) D and basidiospores collected from 10 sporophores, two from each of five different groups (T5); (iv) T1 alone; (v) T5 alone; and (vi) a noninoculated control. Dikaryotic mycelial inoculum was provided at the time of sowing, while basidiospore inoculum was added at 10 weeks after seed germination. Sporophore formation was induced after 20 weeks of growth, and dikaryotic cultures were isolated from their tissue. Seedlings were harvested, and growth and mycorrhization were assessed. Levels of both were generally lower for T1-treated seedlings, compared with seedlings receiving D, while levels for T5-treated seedlings were intermediate. Sporophore genotype variability was assessed for inoculum treatments by using the isoenzymatic marker leucine aminopeptidase. The greatest genetic variability was seen with the basidiospore treatments T1 and T5, with up to four leucine aminopeptidase patterns per seedling. The mixed treatments D plus T1 and D plus T5 produced most frequently, but not exclusively, the inoculated dikaryon genotype. After isoenzyme results were assessed, variable sporophore isolates of mixed treatments were analyzed with randomly amplified polymorphic DNA and PCR mitochondrial DNA markers to determine if they were formed by dikaryon-monokaryon crosses between the inoculated dikaryon and monosporous mycelia.(ABSTRACT TRUNCATED AT 250 WORDS)