Evidence for amphithallism and broad geographical hybridization potential among Agaricus subrufescens isolates from Brazil,France, and Thailand

Agaricus subrufescens is a cultivated edible and medicinal mushroom. Its known geographical distribution encompasses the Americas, Europe, Oceania, and Asia. The objective of this study was to assess mating compatibility and interfertility of strains originating from Brazil, France, and Thailand. Progeny of each strain were analyed with codominant molecular markers. Multilocus genotype tests revealed that the three strains were amphithallic with percentages of heterokaryotic single spore progenies of 75 % for the Thai strain and around 40 % for the Brazilian and French strains. In mating tests A. subrufescens had a multiallelic unifactorial system of sexual incompatibility. The three parent strains were interfertile based on experimental pairings of single-spore isolates, the recovery of hybrid heterokaryons from compatible matings, and the ability of hybrids to produce mushrooms and fertile spores. This biological approach supports the inclusion of the European strains within the species and the extension of the geographical distribution range to Asia. Our data should help to develop breeding strategies and to better manage and exploit the diversity existing in A. subrufescens.     

卵孢侧耳野生菌株人工栽培及营养成分分析

采自西藏日喀则地区的两份野生蘑菇标本及其分离菌株,通过形态学及基于ITS序列的分子系统发育分析鉴定为卵孢侧耳Pleurotus placentodes。该物种在2016年首次报道于中国。人工驯化栽培试验表明卵孢侧耳可以在人工条件下出菇,生长周期80-95d;初步的营养成分分析表明,卵孢侧耳蛋白质、粗多糖、氨基酸含量比香菇、糙皮侧耳（平菇）、金针菇、柱状田头菇（茶树菇）等都高,尤其含有较高的组氨酸和丰富的矿物质,如铁、钙、锌等。卵孢侧耳可作为新的食用菌进行开发利用。     

Developments in breeding of Agaricus bisporus var. bisporus: progress made and technical and legal hurdles to take

True breeding of button mushrooms has hardly been done in the last decades, despite this species being one of the most cultivated mushrooms worldwide. Research done in the last 20 years has identified and characterised new germplasm and improved our understanding of the genetic base for some traits. A substantial collection of wild-collected strains is now available and partly characterised for a number of important traits such as disease resistance and yield. Most of the variations found in a number of important agronomic traits have a considerable heritability and are thus useful for breeding. Genetic marker technology has also developed considerably for this mushrooms in the last decade and used to identify quantitative trait loci (QTL) for important agronomic traits. This progress has, except for one example, not resulted so far into new commercially varieties. One of the reasons lies in the typical life cycle of the button mushroom Agaricus bisporus var. bisporus which hampers breeding. Joint investment is needed to solve technical problems in breeding. Special attention is needed for the protection of new varieties. Due to its typical life cycle, it is very easy to generate so called “look-a-likes” from protected cultivars by screening fertile single spore cultures. A consensus has been reached within the mushroom (breeding) industry to consider this method as the generation of essentially derived varieties as defined in plant breeding.

     

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.     

Development of polymorphic microsatellite markers issued from pyrosequencing technology for the medicinal mushroom Agaricus subrufescens

The recently described procedure of microsatellite-enriched library pyrosequencing was used to isolate microsatellite loci in the gourmet and medicinal mushroom Agaricus subrufescens. Three hundred and five candidate loci containing at least one simple sequence repeats (SSR) locus and for which primers design was successful, were obtained. From a subset of 95 loci, 35 operational and polymorphic SSR markers were developed and characterized on a sample of 14 A.?subrufescens genotypes from diverse origins. These SubSSR markers each displayed from two to 10 alleles with an average of 4.66 alleles per locus. The observed heterozygosity ranged from 0 to 0.71. Several multiplex combinations can be set up, making it possible to genotype up to six markers easily and simultaneously. Cross-amplification in some closely congeneric species was successful for a subset of loci. The 35 microsatellite markers developed here provide a highly valuable molecular tool to study genetic diversity and reproductive biology of A.?subrufescens.     

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.     

Current findings,future trends,and unsolved problems in studies of medicinal mushrooms

The target of the present review is to draw attention to many critically important unsolved problems in the future development of medicinal mushroom science in the twenty-first century. Special attention is paid to mushroom polysaccharides. Many, if not all, higher Basidiomycetes mushrooms contain biologically active polysaccharides in fruit bodies, cultured mycelium, and cultured broth. The data on mushroom polysaccharides are summarized for approximately 700 species of higher Hetero- and Homobasidiomycetes. The chemical structure of polysaccharides and its connection to antitumor activity, including possible ways of chemical modification, experimental testing and clinical use of antitumor or immunostimulating polysaccharides, and possible mechanisms of their biological action, are discussed. Numerous bioactive polysaccharides or polysaccharide–protein complexes from medicinal mushrooms are described that appear to enhance innate and cell-mediated immune responses and exhibit antitumor activities in animals and humans. Stimulation of host immune defense systems by bioactive polymers from medicinal mushrooms has significant effects on the maturation, differentiation, and proliferation of many kinds of immune cells in the host. Many of these mushroom polymers were reported previously to have immunotherapeutic properties by facilitating growth inhibition and destruction of tumor cells. While the mechanism of their antitumor actions is still not completely understood, stimulation and modulation of key host immune responses by these mushroom polymers appears central. Particularly and most importantly for modern medicine are polysaccharides with antitumor and immunostimulating properties. Several of the mushroom polysaccharide compounds have proceeded through phases I, II, and III clinical trials and are used extensively and successfully in Asia to treat various cancers and other diseases. A total of 126 medicinal functions are thought to be produced by medicinal mushrooms and fungi including antitumor, immunomodulating, antioxidant, radical scavenging, cardiovascular, antihypercholesterolemia, antiviral, antibacterial, antiparasitic, antifungal, detoxification, hepatoprotective, and antidiabetic effects.     

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.     

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.     

Simulium (Psilopelmia) virescens, a new black-fly species (Diptera: Simuliidae) from the southwestern region of the state of Bahia, Brazil

The last-instar larva, pupa, male and female of Simulium virescens sp. nov. are described and illustrated. This species has a peculiar larva, which has an elongated head capsule and light-green colour. The first thoracic segment has tubercle on its dorsal region and the third thoracic segment has one pair of tubercles; the first to the fourth abdominal segments have one pair of tubercles on each segment. Until now this new species had only been collected at the type locality, which is on the middle stretch of the Correntina River in the southwestern portion of the state of Bahia, Brazil. Females were voraciously biting humans during the field work. This new species represents the second species of Simulium (Psilopelmia) in Brazil and the first registered outside of the Brazilian Amazon Region.     

Optimization of substrate preparation for oyster mushroom (<Emphasis Type="Italic">Pleurotus ostreatus</Emphasis>) cultivation by studying different raw materials and substrate preparation conditions (composting: phases I and II)

In recent years, oyster mushroom (Pleurotus ostreatus) has become one of the most cultivated mushrooms in the world, mainly in Brazil. Among many factors involved in a mushroom production, substrate preparation is the most critical step, which can be influenced by composting management techniques. Looking forward to optimizing the substrate preparation process, were tested different composting conditions (7 and 14 days of composting with or without conditioning), potential raw materials (decumbens grass, brizantha grass and sugarcane straw) and nitrogen supplementation (with or without wheat bran) on oyster mushroom yield and biological efficiency (BE). The substrate composted for 7 days with conditioning showed higher yield and biological efficiency of mushroom (24.04 and 100.54 %, respectively). Substrates without conditioning (7 and 14 days of composting) showed smaller mushroom yield and biological efficiency. Among the raw materials tested, brizantha grass showed higher mushroom yield followed by decumbens grass, sugarcane straw and wheat straw (28.5, 24.32, 23.5 and 19.27 %, respectively). Brizantha grass also showed higher biological efficiency followed by sugarcane straw, decumbens grass and wheat straw (123.95, 103.70, 96.90 and 86.44 %, respectively). Supplementation with wheat bran improved yield and biological efficiency in all substrate formulations tested; thus, oyster mushroom yield and biological efficiency were influenced by substrate formulation (raw materials), supplementation and composting conditions.     

Diversity and abundance of macrofungi in rubber agroforests in southwestern Nigeria

The mushroom diversity was studied for 14 months in Edo State, southwestern Nigeria. A total of 76 macrofungi tantamount to 333 fruit bodies were encountered, 74% of which were identified and named. The basidiomycetes and members of the family Tricholomataceae were the best represented taxa while species on wood‐based substrates made up 66% of the total mushroom taxa. The monthly mushroom abundance profile, diversity and species richness for each sampled plot correlated with local meteorological factors, age of the sampled sites and distribution of wood‐based substrates. Jaccard, Sorensen and Bray–Curtis similarity index values showed that plots A and D, representing young actively managed and old abandoned rubber tree plantations, respectively, were the most dissimilar in mushroom composition. The hypersensitive response of macrofungal fruit bodies to climate variables qualify mushrooms as veritable biomonitors or bioindicators of climate change. This study supported earlier reports that rubber agroforests are a depauperate pool of utilitarian mushrooms.     

The General Secretory Pathway of Burkholderia gladioli pv. agaricicola BG164R Is Necessary for Cavity Disease in White Button Mushrooms

Cavity disease in white button mushrooms is caused by Burkholderia gladioli pv. agaricicola. We describe the isolation and characterization of six mutants of the strain BG164R that no longer cause this disease on mushrooms. The mutations were mapped to genes of the general secretory pathway (GSP). This is the first report of the association of the type II secretion pathway with a disease in mushrooms. Phenotypes of the six avirulent mutants were the following: an inability to degrade mushroom tissue, a highly reduced capacity to secrete chitinase and protease, and a reduced number of flagella. Using these mutants, we also made the novel observation that the factors causing mushroom tissue degradation, thereby leading to the expression of cavity disease, can be separated from mycelium inhibition because avirulent mutants continued to inhibit the growth of actively growing mushroom mycelia. The GSP locus of B. gladioli was subsequently cloned and mapped and compared to the same locus in closely related species, establishing that the genetic organization of the gsp operon of B. gladioli pv. agaricicola is consistent with that of other species of the genus. We also identify the most common indigenous bacterial population present in the mushroom fruit bodies from a New Zealand farm, one of which, Ewingella americana, was found to be an apparent antagonist of B. gladioli pv. agaricicola. While other investigators have reported enhanced disease symptoms due to interactions between endogenous and disease-causing bacteria in other mushroom diseases, to the best of our knowledge this is the first report of an antagonistic effect.     

担子菌类食用菌交配型位点结构的研究进展

大多数可栽培的食用菌是属于担子菌的大型真菌,具有复杂的交配型系统,通常涉及到两类交配型基因,即编码同源域转录因子的A交配型基因以及编码脂肽信息素和信息素受体的B交配型基因。对担子菌交配型系统的研究已经有上百年的历史,近年来随着高通量测序技术的发展,很多常见食用菌的基因组获得测序,使得我们对不同类型交配型位点的分子遗传学结构能够进行更加细致的解析。本文在概述了担子菌有性生殖系统和交配型基因分子特点的基础上,对常见食用菌中的香菇、金针菇、灵芝、糙皮侧耳、刺芹侧耳、白灵侧耳、裂褶菌、双孢蘑菇、草菇和虎皮香菇以及模式生物灰盖鬼伞等物种的交配型位点的结构进行了总结和分析。从已有的研究结果来看,常见食用菌的交配型位点的分子遗传学结构存在多样性,不同物种的交配型位点具有不同的结构特点。从物种内不同菌株之间的交配型结构比较来看,交配型基因的位置和数量也具有丰富的多样性。在分子遗传学层面对常见食用菌交配型位点结构的认识将有助于深入阐明交配型基因对子实体发育的调控以及解决食用菌生产实际中的科学问题,但是目前对食用菌交配型位点和基因的研究仍旧存在很多空白,有待于进一步深入和拓展。     

Mushroom diversity in sustainable shade tea forest and the effect of fire damage

A survey of the biodiversity of wild macrofungi, including edible species yields, was carried out from 1 May to 30 September 2007 at four different forest types (in mainly Miang tea forest). The plots 100 m², comprised a tea garden with a few planted canopy tree species (37.2% canopy cover), a cultivated sustainable tea forest (80.2% canopy cover), an abandoned sustainable tea forest (89.8% canopy cover), and an abandoned sustainable tea forest that had suffered fire damage (72.9% canopy cover). All visible mushrooms were collected during weekly forays. Macro-characters of the fungi were annotated and photographed by digital camera and the fungi were identified to genera and morphospecies (e.g. Agaricus sp.1, Agaricus sp.2). The fresh weight of wild edible mushrooms produced in the plots was also recorded during this period. The biodiversity of macrofungi in abandoned sustainable tea forest was highest with 115 species in 47 genera, followed by cultivated sustainable tea forest with 85 species in 42 genera and fire damaged abandoned sustainable tea forest with 48 species in 25 genera, while only 19 species belonging to 15 genera were found in the tea garden. Twenty-one species belonging to nine genera with a total of 60.9 kg of edible fungi were collected throughout this study. The fresh weight of edible mushrooms was recorded with the highest fresh weight (25.3 kg) collected from the abandoned sustainable tea forest, followed by 18.2 kg in the cultivated sustainable tea forest, while in the fire damaged sustainable tea forest 16.4 kg were collected and only 1 kg was collected in the tea garden. Abandoned sustainable tea forest contained a greater shade tree biodiversity and higher canopy cover than other plots. This area has a generally higher humidity, a greater ground litter cover, a greater number of microhabitats, a greater number of trees that can form mycorrhizal associations and this probably accounts for the higher diversity of macrofungi and production of edible mushrooms. The fire damaged sustainable tea forest had lower mushroom diversity and edible mushroom production possibly due to the loss of litter and lower tree diversity. The tea garden supported a poor diversity of mushrooms and almost no edible mushrooms. Shade tea forest (Miang tea forest) is a good method to produce tea in a sustainable way as it maintains diversity of mushrooms and other organisms and could be developed as an alternative to shade coffee.     

云南蘑菇中毒事件中的毒蘑菇物种多样性

近年来,云南省每年发生超过500起蘑菇中毒事件,造成2 000余人中毒,约30人死亡,成为我国蘑菇中毒危害最为严重的省份。系统研究蘑菇中毒事件中的毒蘑菇多样性,可以为蘑菇中毒精准防控提供科学依据和技术支撑。本研究对云南省2013年以来开展过科学物种鉴定的223起中毒事件进行了系统分析,发现云南省蘑菇中毒一年四季均有发生,主要集中在6-9月份,中毒事件数和中毒人数7月份达到高峰,而6月份死亡人数最多。地区分布分析发现云南省蘑菇中毒事件涉及14个市州,中毒事件数前5位分别为德宏、玉溪、保山、楚雄和普洱,均位于云南中部及以南地区。223起中毒事件中共成功鉴定出47种毒蘑菇,分属6种中毒类型(急性肝损害型、急性肾衰竭型、横纹肌溶解型、胃肠炎型、神经精神型和光敏性皮炎型)。这些毒蘑菇包含发现于中国的新物种16种,以及2019年以来发现的中国新记录种3种。致命鹅膏Amanita exitialis和亚稀褶红菇Russula subnigricans分别造成19人和9人死亡,是云南省最危险的2种剧毒蘑菇。青褶伞Chlorophyllum molybdites、日本红菇R. japonica、光硬皮马勃Scleroderma cepa、近江粉褶菌Entoloma omiense和发光类脐菇Omphalotus olearius位列胃肠炎型中毒的前5位。热带紫褐裸伞Gymnopilus dilepis、毒歧盖伞近似种Inosperma cf. virosum和兰茂牛肝菌Lanmaoa asiatica位列神经精神型中毒的前3位。叶状耳盘菌Cordierites frondosus是目前云南省造成光敏性皮炎型中毒的唯一物种。     

Double-stranded ribonucleic acid in Agaricus bisporus.

Double-stranded ribonucleic acid present in virus-infected mushrooms of Agaricus bisporus has been resolved through polyacrylamide gel electrophoresis into six molecular-weight forms. Identification of these six double-stranded ribonucleic acids in mushrooms by this procedure has proven to be a useful and diagnostic method for viral infection in the cultivated mushroom.     

Growth and mortality of black bass, Micropterus salmoides (Pisces, Centrachidae; Lacapède, 1802) in a reservoir in southern Brazil.

The black bass, Micropterus salmoides, was introduced to Brazil from North America in 1922. Since then the species has been reared in aquaculture facilities intended to stock reservoirs as additions to native stocks available for angling. At present no scientific information on the biology of black bass in Brazilian waters is available. Since black bass dispersion may cause severe impacts on native Brazilian fish fauna, information on the basic biological parameters of this species is necessary. The objective of the present study is to provide information on the growth, age structure, and mortality of the species in a small reservoir in southern Brazil, where the species reproduces regularly. Based on scale readings, the von Bertalanffy growth curve was calculated and compared to the length-frequency distribution of the population. Both methods showed similar results. Maximum length was about 44 cm. The mean length at the end of the first year was 23.1 cm (s.d. = 13.88); at that of the second, 37.3 cm (s.d. = 12.52); and 41.4 cm (s.d. = 9.92) at the third. Oldest fish were three years old. The growth performance index phi' was 3.28 cm year(-1). Mortality increased from 0.16 year(-1) between the first and the second cohort, to 0.8 year(-1) between the second and third. The results show that black bass in Brazil grows faster than in its area of origin, but longevity is shorter and body shape, stouter. The cause of high mortality at a relatively early age may be connected with the loss of genetic diversity due to inbreeding of the Brazilian stocks, which originated from few introduced individuals a long time ago. The fact that black bass reproduces in reservoirs and grows rapidly may be considered a threat to conserving fish diversity in Brazilian ecosystems.     

Effects of Caenorhabditis elegans (Nematoda: Rhabditidae) on yield and quality of the cultivated mushroom Agaricus bisporus

Thirteen species of saprobic rhabditid nematodes (11 genera) were identified from samples of compost and casing material collected from mushroom farms in the British Isles. Caenorhabditis elegans, the most frequently found saprobe, was mass-produced monoxenically and its effects on the cultivated mushroom, Agaricus bisporus (strain U3) were studied. C. elegans did not multiply in well-prepared, pasteurised, spawned compost, whereas casing material proved to be a highly suitable environment for its reproduction. An initial casing inoculum of 10⁶ nematodes/crate of compost (7.5 kg), caused a significant reduction in mushroom yield. Losses in total mushroom yields of 11%, 20% and 26% were caused by initial inoculum rates of 10⁶, 10⁷and 2 × 10⁷ nematodes/crate, respectively. Yields were negatively correlated with the initial nematode inoculation level and regression equations were derived. The nematode treatments caused fewer mushrooms to be produced and an absence of the usual distinctive flushing patterns. C. elegans caused considerable deterioration in mushroom quality and characteristic distortion of mushrooms. Individual sporophores were mis-shapen, notched and had brown or violet coloured grills. Up to 3.8%, 6.7% and 10.8% of total weight and 3.5%, 5.4% and 8% of total numbers of mushrooms were distorted at the three highest nematode inoculum rates tested. Weights and numbers of distorted mushrooms were positively correlated with the initial nematode population. C. elegans commonly colonised sporophores.