Fruiting Body Production Research Articles

Cultural Characteristics and Fruiting Body Production in Cordyceps bassiana

Single ascospore isolates of Cordyceps bassiana were observed for their colony pigmentation on Sabouraud Dextrose agar plus Yeast Extract (SDAY) plates and were inoculated in a brown rice medium for production of fruiting bodies. Colony pigmentation did not show any relationship with perithecial stromata formation. The isolates were also grown on opposite sides of SDAY agar plates and were observed for vegetative compatibility. Neither vegetative compatibility nor perithecial stromata could be found to be related to each other. It was concluded that fertile fruiting body production was independent of colony pigmentation and vegetative compatibility. Synnemata formation was found to be more common than perithecial stromata formation. This might be due to its highly conidiogenous anamorphic stage, i.e., Beauveria bassiana.

Optimum Conditions for Artificial Fruiting Body Formation of Cordyceps cardinalis

Stromatal fruiting bodies of Cordyceps cardinalis were successfully produced in cereals. Brown rice, German millet and standard millet produced the longest-length of stromata, followed by Chinese pearl barley, Indian millet, black rice and standard barley. Oatmeal produced the shortest-length of fruiting bodies. Supplementation of pupa and larva to the grains resulted in a slightly enhanced production of fruiting bodies; pupa showing better production than larva. 50~60 g of brown rice and 10~20 g of pupa mixed with 50~60 mL of water in 1,000 mL polypropylene (PP) bottle was found to be optimum for fruiting body production. Liquid inoculation of 15~20 mL per PP bottle produced best fruiting bodies. The optimal temperature for the formation of fruiting bodies was 25°C, under conditions of continuous light. Few fruiting bodies were produced under the condition of complete darkness, and the fresh weight was considerable low, compared to that of light condition.

Fruit Body Formation on Silkworm by Cordyceps militaris

Injection inoculation protocols for fruit body formation of Cordyceps militaris (C. militaris) were investigated to improve the incidence of infection in the silkworm species Bombyx mori (B. mori). Injection, with suspensions of C. militaris hyphal bodies into living silkworm pupae, was used to test for fruit body production. Use of Daeseungjam rather than Baegokjam or Keumokjam varieties of B. mori is thought to be suitable for infection by C. militaris. From mounting, nine-day-old to 11-day-old pupae showed the best incidence of infection with a 100 μL injection volume. Silkworm pupae injected with a hyphal suspension concentration of more than 2 × 105 colony-forming unit (cfu) recorded a greater than 96% incidence of infection. Also, fruit bodies of C. militaris were induced and produced at a light intensity between 500 and 1,000 lx.

Opposing developmental functions of Agrocybe aegerita galectin (AAL) during mycelia differentiation

Mycelia of basidiomycetes differentiating into fruiting body is a controlled developmental process, however the underlying molecular mechanism remains unknown. In previous work, a novel fungal Agrocybe aegerita galectin (AAL) was isolated from A. aegerita in our laboratory. AAL was shown to promote mycelial differentiation in A. aegerita and Auricularia polytricha, indicating that AAL might function as a conserved fruiting initiator during basidiomycete mycelia development. In the current work, we investigate the role of AAL in mycelia differentiation and fruiting body formation. First, the expression and localization of AAL in mycelia, primordium and fruiting body were assessed by Western blotting and immunohistochemistry. AAL was found to be ubiquitously expressed in the primordium and fruiting body but not in the mycelia. AAL facilitated mycelia congregation and promoted fruiting body production when AAL was applied on mycelia. At the same time, when AAL was spread on potato dextrose agar (PDA) medium prior to mycelia inoculation, mycelia exhibited slowed growth rates, resulting in mycelia cords formation and inhibition of fruiting body formation. The 5′ regulatory sequence of aal was cloned by ‘genome walking’. Here, we show that aal lack introns in the coding region and the upstream 740 bp sequence was characterized by the existence of core promoter elements, which included: two CCAAT boxes (−535/−280), a GC box (−145), a TATA box (−30) and a fungal leader intron within the 5′ UTR. The identification of regulatory expression elements may provide an explanation to the stage-specific and high-level expression of aal during fruiting development.

Successful Development of Cordyceps bassiana Stromata from Beauveria bassiana

A specimen of Beauveria bassiana was collected from Yang-yang of Gangwon province, Korea in October 2006. Conidial isolates were prepared from the specimen by the dilution method and inoculated in brown rice medium for fruiting body production. After nearly two months incubation for perithecial stromata developed from single isolates as well as from their combinations. They were determined as Cordyceps bassiana by observing the stromatal characters and their conidial structures. This is the first report of the development of C. bassiana from B. bassiana cultures.

La productivité des champignons est-elle favorisée par la croissance des arbres? Résultats d’une expérience d’éclaircie

• Most of the edible forest mushrooms are mycorrhizal and depend on carbohydrates produced by the associated trees. Fruiting patterns of these fungi are not yet fully understood since climatic factors alone do not completely explain mushroom occurrence.• The objective of this study was to retrospectively find out if changing tree growth following an increment thinning has influenced the diversity patterns and productivity of associated forest mushrooms in the fungus reserve La Chaneaz, Switzerland. • The results reveal a clear temporal relationship between the thinning, the growth reaction of trees and the reaction of the fungal community, especially for the ectomycorrhizal species. The tree-ring width of the formerly suppressed beech trees and the fruit body number increased after thinning, leading to a significantly positive correlation between fruit body numbers and tree-ring width.• Fruit body production was influenced by previous annual tree growth, the best accordance was found between fruit body production and the tree-ring width two years previously.• The results support the hypothesis that ectomycorrhizal fruit body production must be linked with the growth of the associated host trees. Moreover, the findings indicate the importance of including mycorrhizal fungi as important players when discussing a tree as a carbon source or sink.

Factors Affecting the Amount of Cadmium Accumulated in Culinary-Medicinal Royal Sun Agaricus, Agaricus brasiliensis S. Wasser et al. (Agaricomycetideae), During Cultivation

This study was conducted to investigate accumulation of heavy metals in Agaricus brasiliensis at commercial farms in Taiwan. Among 14 samples of fruiting bodies of A. brasiliensis examined, five heavy metals, Cr, As, Cd, Hg, and Pb, were detected and only the content of cadmium (Cd) exceeded the level of 2 ppm (μg/g) specified by the Department of Health, Republic of China. The rates of Cd in fruiting bodies varied with samples ranging from 1.10 μg/g to 20.07 μg/g. Accumulation of Cd in fruiting bodies varied with strains of A. brasiliensis. Among the four strains studied, the strain A5 had the lowest rate of Cd absorption (4.57 μg/g) and the highest production of fruiting bodies (4592.4 g/24 bags), making it an ideal strain for commercial production. Among six cultivation adjuvants tested, the oyster shell powder contained the highest level of cadmium. The average cadmium level in the plastic bag was 0.077 μg/g, whereas the cadmium content in oyster shell powder was 0.363 μg/g. This study indicates that the oyster shell powder in the growth substrate may be one of the main sources for the accumulation of cadmium in the fruiting body of A. brasiliensis.

Phenology of epigeous macrofungi found in red gum woodlands

The timing of fruiting body production by epigeous macrofungi is thought to be mostly determined by substrate moisture and temperature. Understanding the environmental cues that influence fruiting can help when designing surveys, interpreting results, and predicting effects of an altered climate. Species fruiting in 22 river red gum ( Eucalyptus camaldulensis) woodland sites in southeastern Australia was recorded at regular intervals over 2 y. Models were constructed to explain the phenology of 25 of the most common species, as well as the total number of species found fruiting on each survey occasion. We found that rainfall minus evaporation and the time of year each influenced fruiting of the common fungi, but to varying degrees depending on species. Using these same variables, the model predictions for the total number of species expected to be found on each survey occasion fit the observations reasonably well ( R 2 = 0.49). The models could be used to estimate the probability of presence for species of conservation interest, to optimise survey timing, or to predict effects of climate change on fruiting.

The origin of mannans found in submerged culture of basidiomycetes

Submerged cultures of basidiomycetes can be used to obtain polysaccharides in a shorter time, when compared to the production and extraction process of mushroom fruiting bodies. This encouraged many authors to develop a variety of culture media to optimize the production of these polymers. In this paper it was observed that the mannans isolated from submerged culture of mushrooms ( Ganoderma lucidum and Pleurotus spp.) are very similar to yeast mannan, suggesting that these polymers could have arisen from yeast extract used in the culture media, and were not being produced by the basidiomycetes. Therefore, when yeast extract is used in the culture media in the submerged culture of basidiomycetes, a part of the fraction assumed as EPS is composed of yeast extract polysaccharides.

Mating tests in Agrocybe cylindracea sensu lato. Recognition of Agrocybe wrightii as a novel species

Mating compatibility tests among seven collections of A. cylindracea (Basidiomycetes, Agaricales) from distant geographic origins were carried out with the aim of determining if A. cylindracea includes one or more taxons from a biological perspective. Fruit body production was used to obtain spore prints, monosporic cultures and to evaluate the fructification potential of compatible crossings. Protoplast isolation, purification and regeneration were used to attempt obtaining monokaryotic isolates from 2-spored specimens. Mating tests evidenced inter-fertility of Asian, American and European strains and the inter-sterility of the Argentinean strain WT-54. These results, in addition to mitochondrial small subunit variable domains phylogeny and morphological data, lead us to consider this strain as a novel species, A. wrightii. The European, Asian and the rest of the American strains belong to A. cylindracea. The concept of A. cylindracea sensu lato includes at least two different species: A. cylindracea and A. wrightii.

Calcareous amendments in truffle culture: A soil nutrition hypothesis

Several statistical studies have indicated that a high concentration of active carbonate in the soil favours Tuber melanosporum fruit body production. The amount of active carbonate is significantly higher and the total carbonate is significantly lower inside than outside the T. melanosporum brûlés. These statistical patterns and other works show that T. melanosporum mycelia may acidify their immediate soil environment and solubilise carbonated fractions. Subsequently, the particular environmental conditions of brûlé soils may favour a secondary carbonate precipitation with a net increase in active carbonate which would in turn favour T. melanosporum mycelia. The greater brûlé size and fruiting are simultaneously both related to how well the fungus is growing, suggesting that the model which best explains the cause–effect of all these observations is a feedback process. Other statistical studies suggest that active carbonate favours increased fruiting of T. melanosporum as compared with Tuber aestivum, Tuber mesentericum and Tuber rufum, which has led to the use of calcareous amendments in truffle culture. We have studied the carbonated fractions of 46 soil samples from T. melanosporum plantations contaminated and uncontaminated with Tuber brumale (a serious problem in truffle culture), and it appears that the use of calcareous amendments does not serve to eradicate T. brumale from these plantations. However, we have observed the positive effect of calcareous amendments on the production of fruit bodies of both species. The similar response by T. melanosporum and T. brumale to soil active carbonate may be related to the close phylogenetic relationships between both species. Based on these findings, we propose a new soil nutrition hypothesis as a guide for research procedures in truffle soil ecology. Soils which are high in active carbonate lead to host plant chlorosis, which is a symptom of an underlying nutrient deficiency. This nutrient deficiency could lead to greater colonisation of T. melanosporum ectomycorrhizas and favour the growth of mycelia, fruit body production and brûlé development, which in turn encourages the formation of new amounts of active carbonate on the basis of the proposed feedback model.

Polyporus tenuiculus: a new naturally occurring mushroom that can be industrially cultivated on agricultural waste

Polyporus tenuiculus is a naturally occurring species from Central and South America that is consumed by different ethnic groups in the region. To determine the optimal conditions for fruiting body production, two strains were assayed on wheat straw and sawdust with or without supplements. Sixty days of incubation at 25 degrees C were needed to produce a solid block. The highest yield was obtained with strain ICFC 383/00 grown on supplemented willow sawdust. In a second experiment the strain ICFC 383/00 and different supplements were used to improve the biological efficiency (BE) and to determine the quality traits and its biodegradation capacity. The highest yields were obtained on sawdust with 25% of supplements reaching 82.7% of BE. Supplements raised the number of flushes, generally from four to five, contributing to increased yields. The type of substrate had a significant effect on fruiting body diameters of P. tenuiculus, and the largest mushrooms were harvested on supplemented substrate with the highest BE coinciding with the highest dry matter loss in substrates. P. tenuiculus showed a capacity to degrade sawdust, causing a decrease of 67.2-74.5% in cellulose, 80.4-85.7% in hemicellulose, and 60.6-66.2% in lignin content at the end of the cultivation cycle. The decrease in hemicellulose was relatively greater than that of cellulose and lignin on supplemented substrates. This is the first report of the cultivation of P. tenuiculus on lignocellulosic waste, and it is a promising species both for commercial production and for its potential use in the degradation of other biowastes.

Biotechnology for in vitro growing of edible and medicinal mushrooms on wood wastes

The aim of this work was focused on finding out the best way to convert the wood wastes into useful food supplements, such as mushroom fruit bodies, by using them as growing sources for the edible and medicinal mushrooms. According to this purpose, three fungal species from Basidiomycetes, namely Ganoderma lucidum (Curt.:Fr.) P. Karst, Lentinus edodes (Berkeley) Pegler and Pleurotus ostreatus (Jacquin ex Fries) Kummer were tested to determine their biological potential to grow on substrates made of wood wastes (sawdusts as well as shavings) which could be used in this way as main ingredients for preparation of natural culture composts. The experiments were achieved by in vitro growing of all these fungal species in special rooms, where the main culture parameters were kept at optimal levels in order to get the highest production of mushroom fruit bodies. The effects of culture compost composition (carbon, nitrogen and mineral sources) as well as other physical and chemical factors (such as: temperature, inoculum amount, pH level and incubation time, etc.) on mycelial net formation and especially on fruit body induction, were investigated. From all these fungal species tested in our experiments, P. ostreatus was registered as the fastest mushroom culture, then L. edodes and finally, G. lucidum as the longest mushroom culture. During the experiments, different logs of the same species were used as control samples for each culture compost variants. Applying such biotechnology, the environmental problems generated by the plant wastes accumulation in wood industry could be solved only by using biological means for their valorising, simultaneously with food supplements producing having high nutritive values as well as healing effects by increasing the consumers` health.

Influence of carbon dioxide, inoculum rate, amount and mixing of casing soil on Agaricus blazei fruiting bodies yield

Influence of carbon dioxide concentration during compost and casing soil overgrowth, inoculum rate, casing soil depth and mixing on Agaricus blazei fruiting bodies yield was determined. Compost composed of chicken manure and wheat straw which is used for commercial button mushrooms cultivation in Slovenia showed to be appropriate for A. blazei fruiting bodies production. Out of the parameters tested in our experiments, casing soil depth and inoculum rate had the biggest positive effect on fruiting bodies production. For A. blazei fruiting bodies production higher rates of inoculum and at least 8.5 kg of commercial casing soil per square meter of compost surface should be used. Higher carbon dioxide concentrations shoved to be beneficial for attaining higher yields of A. blazei fruiting bodies.

Oilseed Rape straw for Cultivation of Oyster Mushroom

AbstractOyster mushroom [Pleurotus ostreatus var. sajor caju (Fr.) Singer] cultivation can play an important role in managing organic waste. It can be cultivated on a wide rang of substrates containing lignin and cellulose. Oyster mushroom was grown on five substrates: Rise straw (Oryza sativa L. var. Alikazemi), Rise straw + Oilseed Rape straw (Brassica napus var. Hyola 401) (75:25 dw /dw), Rise straw + Oilseed Rape straw (50:50 dw /dw), Rise straw + Oilseed Rape straw (25:75 dw /dw) and Oilseed Rape straw alone. Oilseed Rape straw alone and Rise straw + Oilseed Rape straw (25:75 dw /dw) were best for fruit body production of P. ostreatus. Time to fruiting for P. ostreatus was also shorter on Oilseed Rape straw. Protein content of the fruit bodies obtained from Oilseed Rape straw was higher than those from other substrates. Using Oilseed Rape straw as a substrate appears to be suitable for oyster mushroom production.

RECOVERY OF MORE THAN 10 YEARS-DRYING Monascus CULTURES AND ITS PURIFICATION METHODS FROM FUNGAL AND BACTERIAL CONTAMINATION

This study was carried out to understand the recovery capability of more than 10 years- drying Monascus cultures. A new simple purification technique from fungal contamination using ethanol-soaking treatment was also reported as a part of this study. The result showed that all drying cultures were recovered well and retained their characters such as good growth, pigmen­tation and production of fruit bodies (ascomata), sexual spores (ascospores) and asexual spores. Several cultures showed its good growth in 20% ethanol medium. This study also reported suc­cessful purification of cultures from fungal contamination using ethanol-soaking treatment. This self-drying method, therefore, could be suggested as a good long-term preservation method for Monascus cultures. Moreover, purification method from fungal contamination soaked in ethanol 70% or 95% was successfully effective.

Lawn Clippings for Cultivation of Oyster Mushroom

ABSTRACT Oyster mushroom, Pleurotus ostreatus var. sajor caju (Fr.) Singer, can be cultivated on a wide variety of substrates containing lignin and cellulose. Oyster mushroom cultivation can play an important role in managing organic waste. Oyster mushroom was grown on five substrates: sedge (Carex remota L.), lawn clippings (mix of Bermuda grass [Cynodon dactylon (L.) Pers.], ryegrass [Lolium persicum L.], meadow grass [Poa sinaica L.], and fescue [Festuca drymeia L.]), rice (Oryza sativa L. var. Alikazemi) straw, rice straw + sedge, (1:1 wet wt/wet wt), and rice straw + lawn clippings (1:1 wet wt/wet wt). Lawn clippings alone and rice straw + lawn clippings were best for fruit body production of P. ostreatus. Time to fruiting for P. ostreatus was also shorter on lawn clippings. The problem of low yield in the second flush appeared to be overcome by incorporating lawn clippings into the substrate. Protein content of the fruit bodies obtained from sedge and sedge + rice straw were higher than those from other substrates. Lawn clippings, used as a substrate, appear to be suitable for oyster mushroom production. Additional work is needed to determine whether changing the volume of components of the lawn clippings will affect the suitability of the substrate.

Molecular and functional characterization of a Rho GDP dissociation inhibitor in the filamentous fungus Tuber borchii.

BackgroundSmall GTPases of the Rho family function as tightly regulated molecular switches that govern important cellular functions in eukaryotes. Several families of regulatory proteins control their activation cycle and subcellular localization. Members of the guanine nucleotide dissociation inhibitor (GDI) family sequester Rho GTPases from the plasma membrane and keep them in an inactive form.ResultsWe report on the characterization the RhoGDI homolog of Tuber borchii Vittad., an ascomycetous ectomycorrhizal fungus. The Tbgdi gene is present in two copies in the T. borchii genome. The predicted amino acid sequence shows high similarity to other known RhoGDIs. Real time PCR analyses revealed an increased expression of Tbgdi during the phase preparative to the symbiosis instauration, in particular after stimulation with root exudates extracts, that correlates with expression of Tbcdc42. In a translocation assay TbRhoGDI was able to solubilize TbCdc42 from membranes. Surprisingly, TbRhoGDI appeared not to interact with S. cerevisiae Cdc42, precluding the use of yeast as a surrogate model for functional studies. To study the role of TbRhoGDI we performed complementation experiments using a RhoGDI null strain of Dictyostelium discoideum, a model organism where the roles of Rho signaling pathways are well established. For comparison, complementation with mammalian RhoGDI1 and LyGDI was also studied in the null strain. Although interacting with Rac1 isoforms, TbRhoGDI was not able to revert the defects of the D. discoideum RhoGDI null strain, but displayed an additional negative effect on the cAMP-stimulated actin polymerization response.ConclusionT. borchii expresses a functional RhoGDI homolog that appears as an important modulator of cytoskeleton reorganization during polarized apical growth that antecedes symbiosis instauration. The specificity of TbRhoGDI actions was underscored by its inability to elicit a growth defect in S. cerevisiae or to compensate the loss of a D. discoideum RhoGDI. Knowledge of the cell signaling at the basis of cytoskeleton reorganization of ectomycorrhizal fungi is essential for improvements in the production of mycorrhized plant seedlings used in timberland extension programs and fruit body production.

Dynamics of fruit-body production and mycorrhiza formation of ectomycorrhizal ammonia fungi in warm temperate forests in Japan

To investigate the quantitative correlation between dynamics of ectomycorrhiza (ECM) formation and fruit-body production of the ammonia fungi, we treated forest soils in two ECM forests, a Castanopsis cuspidata forest and a Quercus serrata forest, in warm temperate Japan with urea to induce fruiting of ammonia fungi. We identified the ectomycorrhizae of two known species of ammonia fungi, Alnicola lactariolens and Hebeloma vinosophyllum, using morphological typing and PCR-RFLP. ECM initiation, increase, and subsequent decrease preceded the start, increase, and decrease of fruit-body production for each species. We also found many kinds of ECM fungi that did not develop fruit bodies after urea treatment during the observation period, which suggests that it is necessary to expand the definition of ammonia fungi from one that refers solely to the species reproducing on the ground to one which includes those in somatic forms in the soil.

Influence of carbon dioxide, inoculum rate, amount and mixing of casing soil on Agaricus blazei fruiting bodies yield

Influence of carbon dioxide concentration during compost and casing soil overgrowth, inoculum rate, casing soil depth and mixing on Agaricus blazei fruiting bodies yield was determined. Compost composed of chicken manure and wheat straw which is used for commercial button mushrooms cultivation in Slovenia showed to be appropriate for A. blazei fruiting bodies production. Out of the parameters tested in our experiments, casing soil depth and inoculum rate had the biggest positive effect on fruiting bodies production. For A. blazei fruiting bodies production higher rates of inoculum and at least 8.5 kg of commercial casing soil per square meter of compost surface should be used. Higher carbon dioxide concentrations shoved to be beneficial for attaining higher yields of A. blazei fruiting bodies.