Molecular identification of dipteran pests (Diptera: Sciaroidea) from shiitake mushroom

On shiitake farms, mycophagous maggots can cause serious damage by preventing formation of the fruiting body. Recently, these pests have significantly reduced shiitake production in Korea. However, larvae and female adults cannot be identified due to their lack of morphological characteristics. Therefore, farmers and applied entomologists are unable to determine which species is the primary cause of the shiitake damage. In this study, mycophagous flies (colonized larvae) were collected from damaged shiitake farms and subsequently identified by matching identified males with the cytochrome c oxidase subunit I (COI) sequences from the larvae. Divergences of the COI sequences among the species discriminated the clusters clearly, and the mycophagous pests were identified as Camptomyia corticalis and C. heterobia. Interestingly, these two species coexisted under the bark of shiitake oak bed logs.     

Dark winged fungus gnats (Diptera: Sciaridae) collected from shiitake mushroom in Korea

Seven species of sciarid flies were collected in shiitake mushroom farm in Korea. Among them, Lycoriella ingenua (Dufour 1839) and Bradysia difformis Frey 1948 were dominant as possible pests of the shiitake mushroom because the larvae were found on both oak bed logs and in the artificial sawdust beds for shiitake cultivation. Five other species, which were collected in lower numbers, are reported for the first time in Korea: Bradysia longimentula (Sasakawa 1994), Bradysia trispinifera Mohrig & Krivosheina 1979, Leptosciarella (Leptospina) subdentata (Mohrig and Menzel 1992), Scatopsciara camptospina Mohrig and Mamaev 1990, and Xylosciara inornata Mohrig and Krivosheina 1979.     

Characterization of vitamin B12 compounds in the fruiting bodies of shiitake mushroom (Lentinula edodes) and bed logs after fruiting of the mushroom

This study determined the vitamin B₁₂ content in commercially available dried fruiting bodies of shiitake mushroom, Lentinula edodes. The vitamin B₁₂ contents in dried donko-type fruiting bodies with closed caps (5.61 ± 3.90 μg/100 g dry weight), did not significantly differ from those of dried koushin-type fruiting bodies with open caps (4.23 ± 2.42 μg/100 g dry weight). The bed logs after fruiting of the mushroom also contained the vitamin B₁₂ levels similar to that in the dried shiitake fruiting bodies. To determine whether the dried shiitake fruiting bodies and their bed logs contained vitamin B₁₂ or other corrinoid compounds that are inactive in humans, we purified corrinoid compounds using an immunoaffinity column and identified vitamin B₁₂ using vitamin B₁₂-dependent Escherichia coli 215 bioautograms and liquid chromatography-electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) chromatograms. Dried shiitake fruiting bodies rarely contained an unnatural corrinoid vitamin B₁₂[c-lactone] that is inactive in humans. Given that shiitake mushroom lacks the ability to synthesize vitamin B₁₂ de novo, the vitamin B₁₂ found in dried shiitake fruiting bodies must have been derived from the bed logs.     

Mushroom host influence on Lycoriella mali (Diptera: Sciaridae) life cycle

Lycoriella mali Fitch (Diptera: Sciaridae) infests mushroom crops early in the crop cycle. Recent observations in mushroom houses indicated a difference in emergence time and size of adult L. mali developing on various strains of commercial mushrooms. Samples of adult flies from isolated mushroom houses growing Portabella mushrooms were significantly heavier then those from oyster mushroom houses, whereas flies from shiitake mushroom houses were lightest in weight. Flies collected from isolated Portabella mushroom houses were reared on four strains and species of Agaricus and Pleurotus mushrooms. After the adults emerged, females were weighed, mated, and allowed to oviposit. The number of eggs laid increased as the weight of the female increased. Flies collected from isolated Portabella mushroom houses were reared on eight strains and species of mushrooms. Flies were reared for four generations on each host mushroom mycelium then switched to different host mushrooms. Overall, the hybrid strain of Agaricus bisporus (Lange) Imbach (Agaricales: Agaricomycetideae) was the most favorable host for L. mali, whereas the wild strain of A. bisporus was the least favorable host. Mushroom hosts influence developmental time, survivorship, weight, and reproduction of L. mali.     

Levels of physiologically active indole derivatives in the fruiting bodies of some edible mushrooms (Basidiomycota) before and after thermal processing

Indole compounds were found in the fruiting bodies of selected mushroom species both before and after thermal processing. On the basis of HPLC analyses the following indole compounds were detected: l-tryptophan, 5-hydroxytryptophan, 5-methyltryptophan, tryptamine, 5-methyltryptamine, serotonin, indole and melatonin. The compound that was present in the largest amounts in the methanolic extracts from unprocessed mushrooms was 5-hydroxytryptophan in the amount of 7.32 mg/100 g DW in the case of the fruiting bodies of Auricularia polytricha; 15.83 in Suillus bovinus; 22.94 in Macrolepiota procera, and 24.83 in Lentinula edodes. In the methanolic extracts from thermally processed mushrooms the amount of 5-hydroxytryptophan was: 3.52 mg/100 g DW in the case of A. polytricha; 5.65 for Leccinum scabrum and 10.11 for M. procera. In addition, serotonin was found in unprocessed fruiting bodies of two mushroom species: 1.03 mg/100 g DW in L. edodes and 13.99 in L. scabrum, and also in thermally processed mushrooms.     

Occurrence and characterization of injuries caused by Mycotretus apicalis Lacordaire, 1842 (Coleoptera: Erotylidae) on cultivation of Pleurotus sajor-caju

When carrying out experiments on the production of the edible mushroom Pleurotus sajor-caju in the Laboratory of Edible Mushrooms, Universidade Federal de Lavras, Lavras, Brazil, in the second half of 2007, the presence of beetles later identified as belonging to the species Mycotretus apicalis was verified. This is the first recorded instance of this insect in cultures of P. sajor-caju in Brazil. The larvae and adults of this insect feed on the fruiting bodies of commercial harvests, resulting in reduction in mushroom quality. To provide evaluation of the injuries caused by these insects, substrates colonized by P. sajor-caju were infested with 4, 8, 16, 32 and 64 insects per block of substrate being the qualitative and quantitative losses then noted. Despite the lack of an observed decrease in biological efficiency, the injuries caused by these insects affected the commercial quality of the mushrooms, which may result in economic losses. The results showed that infestations of 32 insects per 0.8 kg of substrate led to a depreciation in the prices of mushrooms meant to be sold.     

Non-destructive analysis of the moisture content in shiitake mushrooms (Lentinula edodes) using near-infrared imaging at 1450 nm

Moisture content (MC) is an important factor in the freshness of shiitake mushrooms (Lentinula edodes). In this study, we developed a simple prediction method for MC of shiitake mushrooms using near-infrared (NIR) imaging at 1450 nm, as the band at 1450 nm is strongly correlated with OH groups. First, a NIR image was captured by an InGaAs sensor with a bandpass filter of 1450 nm; the captured image was then calibrated using an image taken at 1100 nm to remove shade and shape effects of mushrooms. The actual MC of shiitake mushrooms was measured using an oven-drying method, and a calibration curve was obtained for the cap surface and the gill side, respectively. The correlation coefficient reached more than 0.85, and the standard error of calibration ranged from 3.1% to 8.2%. Non-destructive MC images were quantified using the constructed calibration curves, and the experimental results showed that differences in the water-holding property can be visualized over 6 h for each side using our method.     

Agaricus subrufescens: A review

Medicinal mushrooms have currently become a hot issue due to their various therapeutic properties. Of these, Agaricus subrufescens, also known as the “almond mushroom”, has long been valued by many societies (i.e., Brazil, China, France, and USA). Since its discovery in 1893, this mushroom has been cultivated throughout the world, especially in Brazil where several strains of A. subrufescens have been developed and used as health food and alternative medicine. This article presents up-to-date information on this mushroom including its taxonomy and health promoting benefits. Medicinal properties of A. subrufescens are emphasized in several studies which are reviewed here. In addition, safety issues concerning the use of this fungus will be discussed.     

Cultivation of the Dible Mushroom Lentinula edodes (Shiitake) in Pasteurized Wheat Straw – Alternative Use of Georthermal Energy in Mexico

Five edible Lentinula edodes strains were evaluated. The mushrooms were cultivated on a wheat straw substrate that was previously pasteurized by immersion in water heated by residual geothermal vapor, which was also used to warm incubation and production rooms. Finely chopped wheat straw (Triticum aestivum L.) was pasteurized and then spawned with supplemented spawn capable of supplying nutrients and enriching the substrate, with the expectation of yield improvement. The samples were incubated for 60 days before the production started and thus, the mushrooms produced had pileus diameters ranging from 5 to 20 cm. The yields fluctuated from 6.2 to 13.9 % (fresh weight of mushrooms/fresh weight of substrate). Biological efficiency ranged from 24.8 to 55.6 % (fresh weight of mushrooms/dry weight of substrate), while the production rate reached varied from 0.19 to 0.55 % (biological efficiency/production time starting from inoculation). The cultivation system evaluated here offers the possibility of lowering production costs by cultivating the mushroom on easily obtainable substrate and shortening the culture cycle. The efficiency of this use of geothermal energy and supplemented spawn for shiitake mushroom cultivation on non‐sterilized substrates was proven.     

Effects of different levels of wheat bran, rice bran and maize powder supplementation with saw dust on the production of shiitake mushroom (Lentinus edodes (Berk.) Singer)

The cultivation of shiitake mushroom (Lentinus edodes) is increasing rapidly in Bangladesh due to its nutritional and medicinal importance with excellent flavor and longer shelf life. With the aim of increased production, we have cultivated L. edodes on saw dust (SD) supplemented with different levels (10%, 15%, 20%, 25%, 30%, 35% and 40%) of wheat bran (WB), rice bran (RB), maize powder (MP) and their combination (WB+RB+MP = 1:1:1) to investigate the growth, yield and quality of this mushroom. Most of the growth, yield and quality parameters varied significantly when mushrooms were cultivated with different levels of supplementation. The yield of mushroom was increased with the level of each supplementation upto a certain level, and then decreased. SD supplemented with 25% WB produced the highest number of fruiting bodies (34.8/500 g packet), highest biological yield (153.3/500 g packet), and biological efficiency (76.6%) of L. edodes. But the yield of the best quality mushroom was observed on SD with 40% WB supplementation; however, the qualities were not always supplementation dose dependent. In this study, we report that 25% WB supplementation with SD may be very effective for higher yield and 40% WB supplementation for better quality of L. edodes.     

Enhanced Yield of Shiitake by Saccharide Amendment of the Synthetic Substrate

Three experiments were performed to determine the effect of selected saccharides on mushroom yield and basidiome size of shiitake (Lentinula edodes) when grown on a synthetic substrate. Substrate formulations of sawdust, wheat bran, and millet were nonamended or amended with sucrose, fructose, or glucose. Addition of sucrose (0.6 to 1.2% [dry weight]) to the substrate stimulated mushroom yield by 11 to 20% or more. Addition of fructose at 1.2% and glucose at 0.6% resulted in similar yield increases. Most of the yield increase occurred on the first break. The substrate amended with 1.2% sucrose tended to have a more synchronous maturation for the second break resulting in fewer days when mushrooms were harvested.     

The inhibitory effects of mushroom extracts on sucrose-dependent oral biofilm formation

Mushrooms contain large quantities of α-glucans. Shiitake (Lentinula edodes), Japan’s most popular edible mushroom, has been reported to contain about 6% (weight/dried weight) of α-(1,3)-glucan. This glucan is one of the major components of oral biofilm formed by the cariogenic bacteria Streptococcus mutans and Streptococcus sobrinus. We found that extracts from shiitake and other edible mushrooms could reduce preformed biofilms of S. mutans and S. sobrinus in the presence of dextranase. We also investigated the α-glucanase activities of shiitake mushroom extracts and their effects on biofilm formation. The extracts possessed α-glucanase activity and degraded water-insoluble glucans from mutans streptococci. The extracts strongly inhibited the sucrose-dependent formation of biofilms by S. mutans and S. sobrinus in the presence of dextranase. Our results suggest that some components of mushrooms, including α-glucanases, might inhibit the sucrose-induced formation of oral biofilms.     

Behavioural response of the fungus gnat,Bradysia impatiens (Diptera: Sciaridae) towards certain edible mushrooms and saprophytic fungi

The fungus gnat Bradysia impatiens (Johannsen) is a worldwide pest inhabiting organic-rich environments, including mushroom-cultivation substrates, which leads to massive production losses of edible mushrooms. To promote a non-insecticidal pest control strategy, we evaluated adult behavioural response towards two saprophytic fungal strains Aspergillus flavus Link and Mucor circinelloides Tiegh., isolated from B. impatiens-inhabiting substrates and six edible mushrooms species. Our results indicated that (a) B. impatiens was most attracted to the oyster mushroom Pleurotus ostreatus among all six edible mushrooms; (b) males and females demonstrated a significant attraction for M. circinelloides over A. flavus and P. ostreatus; (c) adults demonstrated dynamic attraction tendencies that varied with the culture periods of A. flavus and M. circinelloides, wherein females represented most (mean = 78.6%) of the attracted individuals; and (d) larvae that fed on mycelial cultures of M. circinelloides could complete their entire life cycles, but larvae could not survive on a non-fungal or A. flavus diet. These results demonstrate the potential for the development of M. circinelloides in a push-pull strategy to biocontrol this pest in edible mushroom cultivation.     

Effects of the paedogenetic mushroom cecid, Heteropeza pygmaea (Diptera:Cecidomyiidae) on cropping of the cultivated mushroom (Agaricus bisporus)

When introduced into a mushroom crop at rates of 2, 20 or 200 larvae/tray (0.56 m²), the mushroom cecid, Heteropeza pygmaea, caused significant reductions in both yield and number of mushrooms in relation to the infestation level. The reductions were greater when the larvae were introduced at spawning rather than at casing. The yield and number of infested (unmarketable) mushrooms increased significantly in relation to the initial infestation level. Just two H. pygmaea larvae, introduced at spawning, resulted in cecid populations that caused a 12% loss in total yield in addition to a 7% loss due to spoilage. Loss assessment in the future, therefore, should take into account both yield suppression and spoilage. There was little effect of cecid infestation on flush timing and mushroom size was only affected in the fourth flush, when a significant reduction (27%) was shown at the highest infestation rate at spawning.     

Vitamin d(4) in mushrooms

An unknown vitamin D compound was observed in the HPLC-UV chromatogram of edible mushrooms in the course of analyzing vitamin D(2) as part of a food composition study and confirmed by liquid chromatography-mass spectrometry to be vitamin D(4) (22-dihydroergocalciferol). Vitamin D(4) was quantified by HPLC with UV detection, with vitamin [(3)H] itamin D(3) as an internal standard. White button, crimini, portabella, enoki, shiitake, maitake, oyster, morel, chanterelle, and UV-treated portabella mushrooms were analyzed, as four composites each of a total of 71 samples from U.S. retail suppliers and producers. Vitamin D(4) was present (>0.1 μg/100 g) in a total of 18 composites and in at least one composite of each mushroom type except white button. The level was highest in samples with known UV exposure: vitamin D enhanced portabella, and maitake mushrooms from one supplier (0.2-7.0 and 22.5-35.4 μg/100 g, respectively). Other mushrooms had detectable vitamin D(4) in some but not all samples. In one composite of oyster mushrooms the vitamin D(4) content was more than twice that of D(2) (6.29 vs. 2.59 μg/100 g). Vitamin D(4) exceeded 2 μg/100 g in the morel and chanterelle mushroom samples that contained D(4), but was undetectable in two morel samples. The vitamin D(4) precursor 22,23-dihydroergosterol was found in all composites (4.49-16.5 mg/100 g). Vitamin D(4) should be expected to occur in mushrooms exposed to UV light, such as commercially produced vitamin D enhanced products, wild grown mushrooms or other mushrooms receiving incidental exposure. Because vitamin D(4) coeluted with D(3) in the routine HPLC analysis of vitamin D(2) and an alternate mobile phase was necessary for resolution, researchers analyzing vitamin D(2) in mushrooms and using D(3) as an internal standard should verify that the system will resolve vitamins D(3) and D(4).     

Effect of liquid mycelial culture used as a spawn on sawdust cultivation of shiitake (Lentinula edodes)

Mycelium of shiitake (Lentinula edodes) was cultured continuously in liquid medium. The liquid culture was carried out for the production of liquid spawn in the cultivation of this mushroom on synthetic sawdust substrate, and its performance was compared with that of the solid spawn. The initial colonization in culture bags was faster with the solid spawn than with the liquid spawn, but after this stage CO₂ production was higher with the liquid spawn than with the solid spawn. For harvesting sufficient amount of good quality mushrooms, 120 d of incubation in bags was needed with the solid spawn, but this was reduced to 90 d for the sawdust blocks using liquid spawn of less than 50 d old. If continuous culture of the liquid spawn was prolonged over 50 d, immature fruit-bodies or their initials formed during the period of bag incubation. The solid subcultures of the liquid spawns retained the fruiting characteristics acquired in the liquid culture. Liquid culture could be a useful tool for breeding of mushrooms.     

Biodegradation of 2,4-dichlorophenol by shiitake mushroom (Lentinula edodes) using vanillin as an activator

The white-rot shiitake mushroom, Lentinula edodes, was used to degrade an environmentally hazardous compound, 2,4-dichlorophenol (DCP), using vanillin as an activator. Vanillin increased the mycelial growth from 74 to 118 mg/150 ml culture and accelerated laccase and Mn-peroxidase production from the maximum on days 24–28 without vanillin to days 10–14. It eliminated 92 % of 100 mM DCP with 50 mg vanillin/l compared with only 15 % without vanillin. GC–MS revealed that a diaryl ether dimer of DCP was formed in the culture without vanillin, whereas dimer formation was diminished with vanillin addition. This indicates that vanillin enhances the degradation of DCP and disrupts the formation of the toxic dimer. Therefore, lignin-derived phenol such as vanillin can be used as natural and eco-friendly activators to control white-rot mushrooms, thereby facilitating the effective degradation of environmentally hazardous compounds.