Response Surface Analysis of Solid State Growth of <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> Mycelia utilizing Whey Permeate

A novel approach to utilizing whey permeate, the cultivation of mycelia of the edible mushroom Pleurotus ostreatus, is introduced. Response surface analysis (RSA) was successfully applied to determine the combination of substrate concentration, temperature and pH that would result in a maximal mycelial extension rate under solid state cultivation. The conditions to maximize the mycelial extension rate were predicted to be 44 g lactose l⁻¹, pH 6.0 and 24.2 °C. Subsequent verification of these levels agreed with model predictions.     

Protein enrichment and digestibility of soft rush (<Emphasis Type="Italic">Juncus effusus</Emphasis>) and rice residues using edible mushrooms <Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> and <Emphasis Type="Italic">Pleurotus sajor-caju</Emphasis>

Pleurotus species are found to be among the most efficient lignocellulolytic types of white-rot fungi. Rice is the main grain cultivated in the extreme south of Brazil. Defatted rice bran and straw are by-products of low aggregate value. Soft rush (Juncus effusus) is a common native plant also very abundant in the region. In the present work, we evaluated changes in substrate composition after growth of two white-rot fungal species: Pleurotus ostreatus and Pleurotus sajor-caju, aiming to increase protein content and digestibility from substrates through solid fermentations and obtain edible mushrooms of high aggregate value. For that, defatted rice bran, defatted rice straw and soft rush were utilized as substrate. The influence of the variables thermal treatment temperature of substrate, substrate moisture and concentration were evaluated on the protein content, digestibility and biological efficiency. The highest protein enrichment of rice bran in P. sajor-caju-fermented medium was due the fact that there was no fructification in these media, while for the P. ostreatus-fermented medium, part of the synthesized protein was converted into mushrooms. The highest protein enrichments were verified in medium with 80% moisture and 25% soft rush (47.1% using P. ostreatus and 49.0% using P. sajor-caju). A higher digestible protein increase was obtained for both species in media with 70% moisture and 25% soft rush.     

Changes in low molecular weight carbohydrates composition and chitin throughout the cultivation stages ofPleurotus ostreatus

Changes in contents of soluble low molecular weight carbohydrates and chitin in a sawdust-rice bran medium during mycelial growth ofPleurotus ostreatus in bottle cultivation were examined in relation to fruit-body yield of nine stocks. Glucose, mannitol, inositol, sucrose, and trehalose were detected in cultures after mycelial spreading. No significant correlation was observed between contents of soluble low molecular weight carbohydrate during mycelial growth and the fruit-body yield. Negative correlation was found between trehalose content in post-harvest cultures and the fruit-body yield. Chitin content in cultures decreased in the fruiting stage. Positive correlation was detected between chitin content of fruit-bodies and the decrement of chitin in post-harvest culture caused by fruit-body growth.     

Optimization of rice bran and food waste compost contents in mushroom culture medium to maximize mycelial growth rate and fruit body yield of Pleurotus ostreatus

The objective of this study was to find optimum rice bran (RB) and food waste compost (FWC) contents in culture medium for mycelial growth rate and fruit body yield of oyster mushroom, Pleurotus ostreatus. RB (0–20%) and FWC (20–40%) contents significantly affected the fruit body yield of P. ostreatus. Within the design boundaries, the optimum conditions for maximum mycelial growth rate (14.8 cm 20 d⁻¹) were 9.3% RB and 24% FWC. The optimum conditions for maximum fruit body yield (91 g bottle⁻¹) were 12% RB and 25% FWC, which was 153% higher than value (36 ± 11 g bottle⁻¹) at the control condition of 80% w/w sawdust and 20% w/w RB. Our results proved a good potential of FWC to serve as an alternative growth medium for cultivating oyster mushroom, P. ostreatus. This may improve the production efficiency of edible mushrooms to help minimize production costs and maximize farm profits.     

大蒜浸出液对平菇细菌性褐斑病病原菌托拉斯假单胞杆菌的抑菌作用

【背景】由托拉斯假单胞杆菌(Pseudomonas tolaasii)引起的平菇细菌性褐斑病在国内外大面积发生,导致产量降低,并有潜在的安全风险,寻找安全有效的抑菌剂对产业发展具有重要意义。【目的】通过5种不同溶剂提取得到大蒜浸出液,测定其对平菇细菌性褐斑病病原菌托拉斯假单胞杆菌的抑制作用,同时检测其对平菇菌丝生长的作用。【方法】利用抑菌圈法测定5种不同的大蒜浸出液对托拉斯假单胞杆菌的抑菌作用,利用平板扩散法筛选能促进平菇菌丝生长的药剂及适宜的浓度。【结果】5种大蒜浸出液原液对托拉斯假单胞杆菌均有显著的抑菌活性,其中大蒜山杏壳木醋液浸出原液抑菌效果最强。不同浓度的大蒜浸出液抑菌作用比较发现,浓度为10%的大蒜山杏壳木醋液浸出液具有较好的抑菌效果,其抑菌效果与0.33 mg/mL的链霉素相当,并对平菇菌丝生长有显著的促进作用,菌丝生长速度显著大于对照,并且菌丝浓密,边缘整齐。【结论】本研究为大蒜与山杏壳木醋液复配药剂防治平菇细菌性褐斑病奠定了实验基础。     

Can mushrooms fix atmospheric nitrogen?

It is generally reported that fungi likePleurotus spp. can fix nitrogen (N₂). The way they do it is still not clear. The present study hypothesized that only associations of fungi and diazotrophs can fix N₂. This was testedin vitro. Pleurotus ostreatus was inoculated with a bradyrhizobial strain nodulating soybean andP. ostreatus with no inoculation was maintained as a control. At maximum mycelial colonization by the bradyrhizobial strain and biofilm formation, the cultures were subjected to acetylene reduction assay (ARA). Another set of the cultures was evaluated for growth and nitrogen accumulation. Nitrogenase activity was present in the biofilm, but not when the fungus or the bradyrhizobial strain was alone. A significant reduction in mycelial dry weight and a significant increase in nitrogen concentration were observed in the inoculated cultures compared to the controls. The mycelial weight reduction could be attributed to C transfer from the fungus to the bradyrhizobial strain, because of high C cost of biological N₂ fixation. This needs further investigations using¹⁴C isotopic tracers. It is clear from the present study that mushrooms alone cannot fix atmospheric N₂. But when they are in association with diazotrophs, nitrogenase activity is detected because of the diazotrophic N₂ fixation. It is not the fungus that fixes N₂ as reported earlier. Effective N₂ fixing systems, such as the present one, may be used to increase protein content of mushrooms. Our study has implications for future identification of as yet unidentified N₂ systems occurring in the environment.     

Development,persistence and regeneration of foraging ectomycorrhizal mycelial systems in soil microcosms

Development of extraradical mycelia of two strains each of Paxillus involutus and Suillus bovinus in ectomycorrhizal association with Pinus sylvestris seedlings was studied in two dimensions in non-sterile soil microcosms. There were significant inter- and intra-specific differences in extraradical mycelial growth and morphology. The mycelial systems of both strains of P. involutus were diffuse and extended more rapidly than those of S. bovinus. Depending on the strain, P. involutus mycelia were either highly plane filled, with high mass fractal dimension (a measure of space filling) or sparse, low mass fractal dimension systems. Older mycelial systems persisted as linear cords interlinking ectomycorrhizal tips. S. bovinus produced either a mycelium with a mixture of mycelial cords and diffuse fans that rapidly filled explorable area, or a predominately corded mycelium of minimal area cover. In the soil microcosms, mass fractal dimension and mycelial cover tended to increase with time, mycelia encountering litter having significantly greater values. Results are discussed in terms of the ecology of these fungi, their foraging activities and functional importance in forest ecosystems.     

Promoting effect of wood vinegar compounds on fruit-body formation ofPleurotus ostreatus

The promoting effect of wood vinegar compounds on the fruiting ofPleurotus ostreatus (Japanese name, Hiratake) was investigated. Not only crude wood vinegar but its components, 3,5-dimethylphenol, 2-methoxyphenol, butanoic acid and 1-pentanol, had the ability to promote fruit-body formation on liquid medium. For use of these promoters industrially, a test for practical cultivation was carried out using a commercial sawdust medium. The addition of 100 µg/ml butanoic acid and 100 µg/ml 2-methoxyphenol into the sawdust medium after removal of the surface mycelial layer (kinkaki in Japanese) produced 29 and 23% higher yields of fruit-bodies than the control cultures (137.2 g/bottle), respectively. The addition of the crude wood vinegar as a medium component into sawdust substrates in the concentration range of 0.1–6% increased yields of fruit-bodies by 21–42% over the control.     

Glucans from fruit bodies of cultivated mushrooms Pleurotus ostreatus and Pleurotus eryngii: Structure and potential prebiotic activity

Cultivated oyster mushrooms (genus Pleurotus) are interesting as a source of biologically active glucans. Partially, β-glucan from Pleurotus sp. (pleuran) has been used as food supplements due to its immunosuppressive activity. Like other dietary fibre components, oyster mushroom polysaccharides can stimulate the growth of colon microorganisms (probiotics), i.e. act as prebiotics. Specific glucans were isolated from stems of Pleurotus ostreatus and Pleurotus eryngii by subsequent boiling water and alkali extraction. Obtained water soluble (L1), alkali soluble (L2) and insoluble (S) fractions were characterised by various analytical methods. Spectroscopic analysis detected glucans in all the fractions: branched 1,3-1,6-β-d-glucan predominated in L1 and S, while linear 1,3-α-d-glucan in L2. Fractions L1 also contained marked amount of proteins partially in complex with glucans; protein content in L2 was insignificant. Effective deproteinisation of L1 and separation of α- and β-glucans in L2 was achieved by the treatment with phenolic reagent. Small amount of chitin was found in S as a component of cell wall chitin–glucan complex. Potential prebiotic activity of extracts L1 and L2 was testing using nine probiotic strains of Lactobacillus, Bifidobacterium and Enterococcus. These probiotics showed different growth characteristics dependently on used extract and strain specificity due to the presence of structurally diverse compounds. The extracts L1 and L2 can be applied to synbiotic construction only for carefully selected probiotic strains. This exploitation of fruit body extracts extends the use of mushrooms P. ostreatus and P. eryngii for human health.     

果实种类及发育阶段对梨小食心虫幼虫钻蛀率和蛹重的影响

【目的】明确梨小食心虫Grapholitha molesta在寄主果实上的钻蛀率和蛹重与寄主种类和果实发育阶段的关系,为其田间预测预报和综合防控提供技术依据。【方法】在室内条件下测定了接卵后72 h梨小食心虫初孵幼虫在不同发育阶段的3个种共6个品种水果［苹果Malus pumila (嘎啦苹果), 桃Amygdalus persica (大久保毛桃、丰白毛桃和霞光油桃), 梨Pyrus bretschneideri (砀山酥梨和玉露香梨)］果实上的钻蛀率以及在不同果实状态的4个水果品种(大久保毛桃、丰白毛桃、砀山酥梨和玉露香梨)果实上72 h的钻蛀率;比较了接卵后24 h和72 h初孵幼虫在不同发育阶段的3个苹果品种(嘎啦苹果、富士苹果和金冠苹果)果实上的钻蛀率;并测定了不同发育阶段的金冠苹果、富士苹果、嘎啦苹果、砀山酥梨和丰白毛桃上的梨小食心虫蛹重。【结果】接卵后72 h梨小食心虫初孵幼虫在6个不同水果品种果实上的钻蛀率从高到低依次为：霞光油桃＞嘎啦苹果＞砀山酥梨＞玉露香梨＞大久保毛桃＞丰白毛桃;除霞光油桃外,其他5个水果品种果实的发育阶段显著影响初孵幼虫钻蛀率。对于嘎啦苹果,接卵后72 h初孵幼虫在其幼果上的钻蛀率最高(73.69%),在其着色期果实上钻蛀率最低(32.51%);在大久保毛桃、丰白毛桃、砀山酥梨和玉露香梨上,接卵后72 h初孵幼虫的钻蛀率均随果实生长发育而增加。初孵幼虫在3 个苹果品种着色期和成熟期果实上的钻蛀率随处理时间的增加而显著下降,而在其幼果上变化不显著。寄主种类及其发育阶段也显著影响梨小食心虫蛹重,成熟毛桃和梨饲养的雌、雄蛹重显著高于未成熟果实饲养的蛹重,着色期苹果不利于蛹重的增加。【结论】果实种类及发育阶段显著影响梨小食心虫初孵幼虫钻蛀率和蛹重,取食成熟期梨和桃的梨小食心虫初孵幼虫钻蛀率和蛹重显著高于取食未成熟果实的个体。     

5种杀虫剂对黑粪蚊的毒力及对平菇菌丝生长的影响

[目的]黑粪蚊为食用菌生产中的危险害虫之一,通过杀虫剂的毒力测定和对平菇菌丝生长抑制试验,拟筛选获得理想的黑粪蚊防治杀虫剂。[方法]采用毒饵法测定了5种杀虫剂对黑粪蚊幼虫的室内毒力,并研究各杀虫剂对平菇菌菌丝生长的抑制情况。[结果]20%甲氰菊酯乳油、14%阿维·虫螨腈悬浮剂和20%甲维·吡丙醚悬浮剂对黑粪蚊的校正死亡率达80%以上,对黑粪蚊幼虫均有较好的杀灭效果,而苏云金杆菌（以色列亚种）悬浮剂对黑粪蚊幼虫的杀虫效果不理想。5种杀虫剂对平菇菌丝生长均有一定抑制作用,且农药品种间存在极显著差异,20%甲氰菊酯乳油抑制率最高,高浓度480 mg·L^-1处理的抑制率可达16.85%,14%阿维·虫螨腈悬浮剂的抑制率最低,高浓度140 mg·L^-1处理的抑制率仅为5.83%。[结论]本研究中14%阿维·虫螨腈悬浮剂用于平菇菌中黑粪蚊防治效果最好,研究结果将为食用菌黑粪蚊的药剂防治提供理论指导。     

Bioconversion of agricultural lignocellulosic wastes through the cultivation of the edible mushrooms Agrocybe aegerita,Volvariella volvacea and Pleurotus spp.

Ten selected wild and commercial strains of Pleurotus ostreatus,Pleurotus eryngii,Pleurotus pulmonarius, Agrocybe aegerita andVolvariella volvacea were cultivated on three agricultural wastes, i.e. wheat straw (WS), cotton waste (CW) and peanut shells (PS). All species demonstrated significantly higher colonization rates on WS and CW than on PS. WS supported faster growth of A. aegerita and Pleurotus spp., whereas V. volvacea performed better on CW. Comparison of growth rates on composted and non-composted WS and CW substrates revealed that in the latter case faster colonization was achieved, particularly for Pleurotus spp. However, one commercial strain of V. volvacea presented higher growth rates when the composted CW medium was used. Furthermore, earliness in the fructification of P. ostreatus, P. pulmonarius and V. volvacea strains was promoted in CW substrates, while WS favoured earliness of P. eryngii and A. aegerita. Similarly, high sporophore yields were obtained by P. ostreatus and P. pulmonarius on both wastes, whereas WS enhanced yield and basidioma size of P. eryngii and A. aegerita strains and CW production of V. volvacea. The substrates cellulose:lignin ratios were found to be positively correlated to mycelial growth rates and to mushroom yield of P. ostreatus and P. pulmonarius; in addition, positive correlation was also detected for carbon:nitrogen ratio and mushroom yield in P. eryngii and A. aegerita and between cellulose content and mushroom yield for V. volvacea strains.     

Cultivation of the oyster mushroom (Pleurotus ostreatus) on wood substrates in Hawaii

Summary Five non-native, aggressively growing trees, Falcataria moluccana (Miquel) Barneby & Grimes, Casuarina equisetifolia L. ex J. R. & G. Forst, Eucalyptus grandis Hill ex Maid, Psidium cattleianum Sabine, and Trema orientalis (L.) Blume, were evaluated for suitability as substrate for outdoor cultivation of the oyster mushroom, Pleurotus ostreatus (Jacq.: Fr.) Kumm., in Hawaii. An existing shade house was modified for mushroom production and proved to be an adequate fruiting site. Nitrogen-fixing trees (C. equisetifolia, T. orientalis, and F. moluccana) supported greater yield (275.5, 272.4 and 268.8 g/bag, respectively), biological efficiency (70.1, 78.5, and 74.0%, respectively), and flush number (3.0, 3.2, and 3.5) than non-fixers. P. cattleianum supported significantly lower yield (190.5 g/bag) and biological efficiency (44.2%). Mean crop period was 51 days and was not affected by the wood substrate. Similarly, substrate did not have a significant impact on the concentration of nutrients or moisture in fruit bodies. Taste preferences were noted in mushrooms grown on different substrates; those grown on C. equisetifolia were most flavourful and preferred in one taste test.     

Larval performance of the oriental fruit moth across fruits from primary and secondary hosts

A common characteristic of many invasive herbivorous insects is their ability to utilize a broad range of host plants. By using various hosts in phenological succession, multivoltine herbivores may increase the number of successful annual generations, at the same time as potentially increasing their overall fitness. To achieve such success, herbivores must be able to develop efficiently on the nutritional resources offered by their hosts. The oriental fruit moth Cydia (= Grapholita) molesta (Busck) (Lepidoptera: Tortricidae) is one of the most damaging invasive insect species. Peach (Prunus persica) is its primary host, whereas the pome fruits apple (Malus × domestica Borkh) and pear (Pyrus communis) are considered as secondary hosts. In many parts of its geographical range, including southern Europe, populations of the moth switch from peach to apple or pear orchards during the growing season. The present study tests whether this temporal switch is supported by the physiological capability of the larvae with respect to developing efficiently on fruits of these taxonomically‐related host plants. Larvae are reared on peach, apple or pear fruits; several life‐history traits are measured; and correlations between the traits are calculated. The results obtained show that larvae do not have the same physiological capability with respect to using apple or pear fruits as hosts compared with using peach fruit. Pear fruit in particular is a sub‐optimal diet. These findings suggest that, in the case of continuous geographical expansion, concomitantly with global warming, apple orchards might support oriental fruit moth populations better than pear orchards, and that the switch onto novel hosts might be accompanied by restricted population growth.     

Molecular genetic characterization of bacterial isolates causing brown blotch on cultivated mushrooms in Japan

The polymerase chain-reaction (PCR) was used to amplify 16S ribosomal DNA (16S rDNA) from bacteria, identified asPseudomonas tolaasii orP. fluorescens, causing brown blotch on cultivated mushrooms in Japan. PCR-amplified 16S rDNA was analyzed on the basis of nucleotide sequence and restriction fragment length polymorphisms (RFLP) to determine the specific identity of isolates. Banding patterns obtained through PCR using primers corresponding to repetitive extragenic palindromic sequences of enteric bacteria (REP-PCR) were used to determine the relatedness of conspecific isolates. AllP. tolaasii isolates and a mushroom pathogen identified asP. fluorescens had identical RFLP patterns and partial 16S sequences, and are considered conspecific. An isolate ofP. fluorescens from creamery wastes (IFO 3507) differed slightly from isolates ofP. tolaasii in both 16S sequence (0.8%) and RFLP patterns (d=0.08), and had almost entirely different REP-PCR bands (d=0.88–1.0). Phylogenetic analyses based on 16S sequences indicated thatP. tolaasii andP. fluorescens are close members ofPseudomonas sensu stricto. REP-PCR shows promise in characterizing isolates pathogenic on different mushroom crops. Two isolates ofP. tolaasii pathogenic onPleurotus ostreatus had identical banding patterns, but three isolates fromLentinula edodes showed the greatest diversity. Contribution No. 312 of the Tottori Mycological Institute, Totori, Japan.     

Fungus fruit body lytic enzyme from a myxomycete,Badhamia utricularis

Chitinase,β-1,3-glucanase, cellulase, xylanase and protease activity were detected in a crude enzyme preparation obtained from a slime mold (Badhamia utricularis) which was grown on autoclaved mycelia ofPholiota nameko in a petri dish. The optimal pH of the enzyme preparation for lytic activity against fruit bodies ofLentinus edodes was 4.0, and those ofβ-1,3-glucanase and cellulase were the same. On the other hand, chitinase and protease showed optimal activity at pH 5.0 and 8.0, respectively. The lytic activity was stable below 40°C but completely inactivated at 70°C, and was most stable at pH 5.0. The studies of the optimal pH, thermal stability, and pH stability, and isoelectric focusing analysis of the enzyme preparation suggest that chitinase,β-1,3-glucanase and cellulase activities may be responsible for lysis of fruit bodies of some mushrooms. The crude enzyme preparation from the slime mold lysed fruit bodies of several mushrooms more efficiently than did commercial lytic enzymes preparations (Driselase and Usukizyme).     

Cultural characteristics and extraction of the fungal pigment phleichrome from the phytopathogenic fungus<Emphasis Type="Italic">Cladosporium phlei</Emphasis>

The cultural characteristics of the fungusCladosporium phlei were assessed in order to develop an improved method for the production of the fungal pigment, phleichrome, which is an intermediate in the production of a photodynamic therapeutic agent. The growth ofC. phlei, as measured by the hyphal growth rate and increase in biomass, varies significantly depending on the culture media utilized (V8 juice-based medium proved optimal for both growth rate and biomass increase). How-ever, even on a V8 juice plate, the growth ofC. phlei occurred slowly and in a limited fashion, in that the colony covered only 75% of the agar surface after more than 4 weeks of cultivation at 20°C. Supplementations of glucose, fructose, galactose, and sucrose increased both hyphal expansion and mass production, whereas supplementations of other carbon sources, including glycerol and sorbitol, exerted no detectable effects. The effect of inorganic nitrogen supplementation was negligible, whereas organic nitrogen evidenced significant effects, with enhanced growth with malt extract and growth inhibition with yeast extract and tryptone. Sporulation was enhanced under conditions of continuous light, and a minimum of 10³ spores per mL of liquid media was found to be necessary for the optimal mass increase. A simple extraction procedure was established in order to isolate the deep red pigment which was subsequently identified as phleichrome via NMR analysis. WhenC. phlei was cultured on V8 medium containing 5% glucose and 2% malt extract, the quantity of mycelial mass was estimated as 20.6 g (dry weight) per liter of culture. The expected phleichrome yields from the mycelia and culture filtrates were estimated to be 43 and 2 mg/L, repectively. There was an equal contribution of the reported research by the first two authors.     

Cultivation of Pleurotus ostreatus and other edible mushrooms

Pleurotus ostreatus is the second most cultivated edible mushroom worldwide after Agaricus bisporus. It has economic and ecological values and medicinal properties. Mushroom culture has moved toward diversification with the production of other mushrooms. Edible mushrooms are able to colonize and degrade a large variety of lignocellulosic substrates and other wastes which are produced primarily through the activities of the agricultural, forest, and food-processing industries. Particularly, P. ostreatus requires a shorter growth time in comparison to other edible mushrooms. The substrate used for their cultivation does not require sterilization, only pasteurization, which is less expensive. Growing oyster mushrooms convert a high percentage of the substrate to fruiting bodies, increasing profitability. P. ostreatus demands few environmental controls, and their fruiting bodies are not often attacked by diseases and pests, and they can be cultivated in a simple and cheap way. All this makes P. ostreatus cultivation an excellent alternative for production of mushrooms when compared to other mushrooms.     

Host feeding,damage and control of the mushroom pest,Brennandania lambi (Acari: Pygmephoroidea) in China

The pygmephoroid mite, Brennandania lambi (Krczal), is the most serious mite pest of mushroom production in Shanghai. China. This mite could not develop and reproduce on the mycelia of Lentinus edodes, Flammulina velutipes, Pleurotus ostreatus or Pleutotus sajor-caju, but 12–20% of the females survived 15 to 31 days on these hosts, whereas all mites died after 8 days in absence of food. When fed the mycelia of Auricularia polytricha, B. lambi adults could become gravid and reproduce but active larvae failed to develop to the next stage. B. lambi developed and reproduced on the mycelia of Auricularia auricula, Hericium erinaceus, Tremella fuciformis and Agaricus bisporus. Mobile stages of B. lambi damage A. bisporus by feeding on its mycelia. Field surveys of mite damage during 1983–1989 in the suburbs of Shanghai revealed that contaminated spawn was the major source of the mite problem and losses of the crop averaged between 10% and 20%. Experiments on the effects of temperature on mite survival indicate that exposure of mites at 50° C for 1 hour or-10°C for 24 hours killed all stages of B. lambi. Thus, heating treatment during well-executed phase II composting and freezing of mushroom spawn at-10° C for 24 hours should eliminate or at least reduce the damage to mushrooms by this pest.