Molecular characterization of Morchella species from the Western Himalayan region of India

The molecular diversity of thirty-two different Morchella cultures/fruiting bodies, collected from the Western Himalayan region was studied in this investigation. Considerable taxonomic confusion exists regarding many species of Morchella. Although classical taxonomy is helpful in identification for many ascomycetes, morels exhibit considerable morphological diversity and there is disagreement in the identification of morel species. Phylogenetic analyses based on DNA sequences could help in sorting out morel taxonomy which is essential to better define the morel diversity. In this study, sequence analysis revealed that in the Western Himalayan region of India, both yellow (M. crassipes, M. spongiola) and black morels (M. elata, M. angusticeps, and M. gigas) were prominent along with two Verpa species. Phylogenetic analysis by maximum parsimony, maximum likelihood and Bayesian inference revealed two different clades and a clear distinction between yellow and black morels.     

Hybridization,characterization and transferability of SSRs in the genus Morchella

Recently, Morchella importuna, M. sextelata, M. eximia, M. exuberans, Mel-13, and Mel-21 have been successfully cultivated in China and planting areas rapidly expanded because of their economic importance. Effective molecular markers are urgently needed for accurately identifying morel cultivars. Microsatellites are widely utilized for strain authentication in many fungal groups. To our knowledge, for the first time we characterized the distribution of microsatellites (simple sequence repeats, SSRs) in the M. importuna genome with 12902 SSRs and reported the first set of SSRs developed for Morchella species. Mono-nucleotides (66.2 %) were the most frequent motifs, followed by tri- (15.4 %), di- (12.1 %), tetra- (3.7 %), penta- (1.3 %) and hexa-nucleotides (1.3 %). We tested the cross-species amplification of 180 SSRs on 24 samples from the six species and high cross-species transferability of SSRs (87.7 %) was found. Among twenty-two microsatellites selected for genetic diversity analysis on 127 samples from the six species, fifteen to twenty polymorphic loci were identified in M. importuna, M. sextelata, M. eximia, M. exuberans, Mel-13 and Mel-21. Interspecific hybridization events were detected among morel species, indicating the potential application of morel crossbreeding. Ninety-one cultivated samples were characterized as new cultivars with different genotypes, but cultivar names used for these by farmers was confusing, with misnaming, synonyms and homonyms. Our results are not only helpful for cultivar identification and morel breeding programs in China, but also provide molecular tools for genetic studies in morels.     

Using Local Ecological Knowledge to Assess Morel Decline in the U.S. Mid–Atlantic Region

Using Local Ecological Knowledge to Assess Morel Decline in the U.S. Mid–Atlantic Region. Morels (Morchella spp.) are prized wild edible mushrooms. In the United States, morels are the focus of family traditions, local festivals, mycological society forays, and social media, as well as substantial commercial trade. A majority of the anglophone research on morels has been conducted in Europe and in the U.S. Pacific Northwest and Midwest. This literature provides insights into a diverse and plastic genus, but much of its biology and ecology remains a mystery. In 2004, we initiated a study of morel mushroom harvesting in the U.S. Mid–Atlantic region in response to concerns that morels might be in decline in the national parks in that area. This paper presents results from that research with an emphasis on morel hunters’ local ecological knowledge of morel types, phenology, habitat, vegetative associations, and responses to disturbance. We conclude that experienced morel harvesters possess local ecological knowledge that complements scientific knowledge and can increase our understanding of the complex and regionally variable ecology of Morchella and inform conservation efforts.     

Morel Phylogeny and Diagnostics Based on Restriction Fragment Length Polymorphism Analysis of ITS Region of 5.8S Ribosomal RNA Gene

Thirty-six cultures representing eight Morchella and related genera, namely, Morchella esculenta, M. crassipes, M. spongiola, M. vulgaris, M. angusticeps, M. conica, Mitrophora semilibera and Verpa conica were subjected to restriction analysis of ITS1-5.8SITS2 region of rDNA. Six restriction endonuclease enzymes viz TaqI, EcoRl, Mspl, Rsal, Hinfl and BsuRl were used to generate restriction fragments and analysis of phylogenetic relationships among morels. The Amplified Ribosomal DNA Restriction Analysis (ARDRA) not only distinguished yellow morels from black morels but also separated related genera Mitrophora semilibera and Verpa conica from true morels. Simultaneously, each morel species could be separated from each other exhibiting considerable phylogenetic distances. The unique restriction fragment profiles generated by the restriction endonucleases enabled us to identify marker fragments to distinguish each species within and amongst the morel group. Since no intra-specific variation in restriction profiles by the six restriction endonucleases could be visualized among monospores, the technique could be used for rapid identification of wild morel specimens as a cheap alternative to direct sequencing for germplasm cataloguing.     

Biotechnology of morel mushrooms: successful fruiting body formation and development in a soilless system

Morchella spp. ascocarps (morels) are some of the world’s most sought-after mushrooms. Successful cultivation of morels is still a rare and difficult task despite over 100 years of effort. Here we provide the first report of successful Morchella rufobrunnea fruiting body initiation and development in laboratory-scale experiments. Mushroom initials appeared 2 to 4 weeks after first watering of pre-grown sclerotia incubated at 16 to 22°C and 90% humidity. Mature fruiting bodies reached 7 to 15 cm in length and were obtained after the five morphological developmental stages of this Morchella species: sclerotium formation, scelerotium germination, asexual spore formation, formation of initial knots and development of the fruiting body.     

Observations on post-fire black morel ascocarp development in an Israeli burnt forest site and their preferred micro-sites

Morchella species ascocarps (morels) are sought-after edible mushrooms that exhibit pyrophilous behavior, proliferating in fire-affected soils of certain types of forests. The factors governing fruiting in this habitat are poorly understood. An observational approach was used to determine the spatial distribution on preferred micro-sites of black morel fruiting in a forest after a summer fire, subjected to different post-fire forestry management activities. Clearing the burnt tree stumps from the site, compaction of the burnt soil by heavy machinery (bulldozers) and covering the soil with chopped wood created preferred micro-sites for black morel fruiting. Fewer fruit bodies developed on untouched burnt soil, and almost none on non-burnt soil at the same site. These observations enhance understanding the ecological principles underlying the distribution and abundance of morel ascocarp development in natural habitats; such an understanding could contribute to conservation and management of morel fruiting in the wild.     

Characterization of mating-type idiomorphs suggests that <Emphasis Type="Italic">Morchella importuna,Mel-20</Emphasis> and <Emphasis Type="Italic">M. sextelata</Emphasis> are heterothallic

Morels (Morchella spp.) are highly prized for their culinary qualities and intensively collected worldwide by mycophiles. Morels are divided into three clades by phylogenetic analyses: black morels, yellow morels and the rufobrunnea clade. Morchella importuna, Mel-20 and M. sextelata are included in the black morel clade and are widely distributed in Yunnan province, China. M. importuna and M. sextelata have been artificially cultured in recent years, but their life cycles and reproductive systems are still poorly understood, which delays the progress of morel cultivation. In this study, the genomes of two ascospore isolates of M. importuna with opposite mating-type were sequenced and two idiomorphs, MAT1–1 and MAT1–2, were identified. The MAT1–2 idiomorph was 6.7 kb in length containing a single MAT1–2-1 gene, and the MAT1–1 idiomorph was 10.5 kb containing a MAT1–1-1 gene and two other open reading frames (ORFs), GME3123 and GME3124. These ORFs differed greatly from the homologues of previously published mating-type genes; therefore, we speculate that they are novel mating genes found only in morels. Single-ascospore populations of M. importuna, M. sextelata and Mel-20 were analysed, and the result indicated that the ratios of MAT1–1- and MAT1–2-harbouring idiomorphs were not significantly different from a 1:1 ratio. The results suggest that these three black morels are heterothallic.     

羊肚菌烂柄病发生对土壤真菌群落结构的影响

为了解羊肚菌(Morel)烂柄病的发生对土壤真菌群落结构的影响,采用Illumina MiSeq高通量测序技术,对健康羊肚菌根际土、烂柄病发病子实体根际土及相同环境下未栽培羊肚菌土壤的真菌群落结构进行研究。结果表明,测序样品共获得344 163条序列,归为7个真菌门。各样品真菌多样性分析结果表明,未栽培土壤真菌群落多样性较高,栽培羊肚菌根际土壤真菌多样性显著降低,烂柄病发生后土壤真菌多样性增加。群落结构分析表明,烂柄病根际优势真菌类群为拟青霉属(Paecilomyces)、木霉属(Trichoderma)、葡萄穗霉属(Stachybotrys)、枝顶孢属(Acremonium)、Paratritirachium、Zopfiella、被孢霉属(Mortierella)和柄孢壳属(Podospora)。烂柄病的发生改变了土壤真菌群落结构,促进了根际真菌的繁殖。为了解羊肚菌烂柄病的发生、传播机理及防治提供了参考。     

甘肃省野生羊肚菌根际细菌群落与土壤环境因子相关性研究

[背景]羊肚菌是全球广泛分布的物种,具有重要的经济和科研价值,其根际微生态系统各要素间的相关性研究相对较少.[目的]探究甘肃省不同地区野生羊肚菌根际土壤中细菌群落-土壤理化性质及细菌群落-酶活性相关性.[方法]采用Illumina MiSeq高通量测序技术,对细菌群落组成进行测量,进而分析其多样性,最终揭示细菌群落-土...     

Effects of different cultivation material formulas on the growth and quality of Morchella spp.

To study the effects of different cultivation material formulas on the growth and quality of Morchella spp. With the cultivated species strains extracted from wild Morchella spp. in Diebu County, Gannan Prefecture as experimental materials, an experiment was designed and the data then obtained was anyalyzed using the single factor variable method. By measuring the pileus length, pileus perimeter, stipe length, stipe perimeter and yield as well as the ash content, total sugar content, crude protein content and crude fiber content of wild Morchella spp., the effects of four different cultivation material formulas on the growth and quality of Morchella spp. were studied. The result showed that the Morchella spp. cultivated using Formula 1, i.e., the formula to which Morchella spp. footing soil was added, grew best, and had the highest yield and the best quality; and the qualities of Morchella spps cultivated using other formulas decreased in a row. Formula 1 to which Morchella spp. footing soil was added had the optimal effect on promoting the growth and quality of Morchella spp.     

DNA polymorphism in morels: complete sequences of the internal transcribed spacer of genes coding for rRNA in Morchella esculenta (yellow morel) and Morchella conica (black morel).

The internal transcribed spacer (ITS) of the gene coding for rRNA was sequenced in both directions with the gene walking technique in a black morel (Morchella conica) and a yellow morel (M. esculenta) to elucidate the ITS length discrepancy between the two species groups (750-bp ITS in black morels and 1,150-bp ITS in yellow morels.     

Ecological characterization of Morel (Morchella spp.) habitats: A multivariate comparison from three forest types of district Swat,Pakistan

Morchella spp. are wild edible mushrooms growing ephemerally under particular environmental conditions in temperate regions of the world. The present study was aimed to outline some principal eco-edaphic characteristics influencing distribution of these mushrooms from three different forest types of Swat, Pakistan. A randomized circular plots were laid down along the transect line in 14 stands in study areas. Environmental variables were recorded and soil characteristics were determined, analyzed and compared. Five species were collected from N to NW slope at elevation range of 1100-2695m asl. Mean relative humidity of 58.7%, range of air temperature 13 °C-27 °C and soil temperature 6-26 °C were recorded at morel growing sites. A higher canopy cover (average 57%) favours sporocarp formation in morels. Soil analysis showed that morel preferably appeared in sandy loam to loamy textured soil at slightly acidic to neutral pH (mean=6.4) and with consistently higher organic matter (1.5%) and salt activities (mean = 38.8 μS) in the soil. IVI calculation showed that Pinus wallichiana A. B. Jackson, Juglans regia L., Abies pindrow Royle, Cedrus deodara (Roxb. ex D. Don) G. Don were dominant trees and Viburnum grandiflorum Wall. ex DC, Hedra nepalensis K. Koch, Hert and Sarcococca saligna (D. Don) Muell. Arg. were shrub species associated with morels. Whilst Fragaria nubicola (Hook.f.) Lindl., Viola canescens Wall ex Roxb. and Podophyllum hexandrum Roylae were the predominant herbs in morel habitat. This study will help to understand the natural habitat and to provide a base line information for the future conservation and management planning in the area.     

近十年中国重要食药用菌研究进展

食用菌跻身我国第五大农作物,我国已经成为名副其实的食药用菌大国。本文对近十年来食药用菌重要种类名称变更、人工培植或栽培技术的突破和基因组学研究的进展进行了综述。随着分子系统学研究的深入,灵芝、黑木耳、毛木耳、桑黄等重要种类的名称得到更正和完善;冬虫夏草、羊肚菌、暗褐网柄牛肝菌等难栽培种类在全世界范围内首次实现人工栽培;大多数常见食药用菌种类完成了基因组解析。分析了分类和系统发育研究、生物学研究、组学研究与食药用菌产业发展的关系,基础研究促进食药用菌产业发展,同时产业发展为基础研究提出更多的科学问题。     

A multigene molecular phylogenetic assessment of true morels (Morchella) in Turkey

A collection of 247 true morels (Morchella spp.) primarily from the Mediterranean and Aegean Regions of Southern Turkey, were analyzed for species diversity using partial RNA polymerase I (RPB1) and nuclear ribosomal large subunit (LSU) rDNA gene sequences. Based on the result of this initial screen, 62 collections representing the full range of genetic diversity sampled were subjected to multigene phylogenetic species recognition based on genealogical concordance (GCPSR). The 62-taxon dataset consisted of partial sequences from three nuclear protein-coding genes, RNA polymerase I (RPB1), RNA polymerase II (RPB2), translation elongation factor (EF1-α), and partial LSU rDNA gene sequences. Phylogenetic analyses of the individual and combined datasets, using maximum parsimony (MP) and maximum likelihood (ML), yielded nearly fully resolved phylogenies that were highly concordant topologically. GCPSR analysis of the 62-taxon dataset resolved 15 putative phylogenetically distinct species. The early diverging Elata (black morels) and Esculenta Clades (yellow morels) were represented, respectively, by 13 and two species. Because a Latin binomial can be applied with confidence to only one of the 15 species (Morchella semilibera), species were identified by clade (Mel for Elata and Mes for Esculenta) followed by a unique Arabic number for each species within these two clades. Eight of the species within the Elata Clade appear to be novel, including all seven species within the Mel-20-to-31 subclade and its sister designated Mel-25. Results of the present study provide essential data for ensuring the sustainability of morel harvests through the formulation of sound conservation policies.     

Successful induction and recognition of conidiation,conidial germination and chlamydospore formation in pure culture of Morchella

Morels, fungi from the genus Morchella, are popular edible mushrooms. However, little knowledge of their asexual reproduction and inaccessible pure mitospores hamper illumination of their life cycle. Herein, we successfully induced conidiation, conidial germination and chlamydospore formation in pure culture of Morchella sextelata. Conidiation proceeded via four morphologically distinct stages: development of the conidiophore stalk, stalk branching, phialide differentiation, and conidium production. Terminal and intercalary chlamydospores were formed on conidial hyphae. The development of conidiophores occurred earlier, with more conidia produced, in cross-mating cultures than in single-spore cultures. Mature conidia were spherical and 2.5–8 μm in diameter, with a vast majority (nearly 99%) 2.5–5 μm in diameter. Each conidium contained one to three nuclei (80.2% conidia contained one nucleus, 19.1% contained two nuclei, and 0.7% contained three nuclei). The conidial nucleus diameter was 1–2 μm. The nuclear mitosis in detached conidia that was observed may benefit their colony initiation. Additionally, morel conidia formed conidial anastomosis tubes. Conidia (mitospores) likely not only function as spermatia, but also as reproductive propagules in Morchella. Further research is imperative to elucidate the relationship between the conidia and chlamydospores, and their unique function in the morel life cycle.     

梯棱羊肚菌外源营养作用的影响因素

外源营养袋的应用是国内羊肚菌大田栽培成功的关键技术之一,但其作用机理和影响因素一直没有得到充分的解析。本研究首先于室内建立一个外源营养模型,确认模型中外源营养模块可以向外输出营养,且被梯棱羊肚菌菌丝利用。通过模型实验发现,外源营养添加到贫瘠培养基时,菌体生物量的增加显著高于营养丰富的培养基;在梯棱羊肚菌生长的不同阶段添加外源营养,菌核形成位置不同,并影响生物量的增加,在菌核起始阶段添加,主培养基的生物量增加最多;外源营养的添加位置、块数等因素在本模型中没有检测到对生物量的影响。外源营养使用聚丙烯袋包裹和不包裹比较发现,包裹组的主培养基内生物量增速和总量低于无包裹组,而外源营养块内趋势相反,但最终两组之间外源营养块与主培养基内生物量总和没有显著差异。使用滤纸和铝箔包裹外源营养时,对生物量的影响和聚丙烯袋材质没有显著差别。外源营养碳氮比20:1时,最有利于主培养基生物量的积累。本研究建立的外源营养研究模型得到的结论与大田栽培有较好的一致性,通过模型发现了实际栽培中不容易观察到的现象,有助于对外源营养袋作用机理进行深入研究。     

Arbitrary Primer Based RAPD — A Useful Genetic Marker for Species Identification in Morels

Internal transcribed spacer (ITS) regions of 5.8S ribosomal DNA gene of 46 monospore cultures representing different species of family Morchellaceae were sequenced. Eight putative species namely Morchella esculenta, M. crassipes, M. angusticeps, M. conice, Mitrophora semilibera, Morchella spongiola, M. vulgaris and Verpa conica were designated to the sequenced cultures analyzed based on similarity with the best aligned sequence of the BLAST search. We recorded interspecific polymorphism in ITS regions of the 5.8S r-DNA gene amongst putative species of morels. However no intraspecific ITS polymorphism could be visualized amongst monospores from single and or different ascocarps collected from various geographical regions. PCR RAPDamplified profiles of different monospores were identical at intraspecific levels and could not be distinguished by eight arbitrary primers tested. RAPD profiles exhibited significant interspecific polymorphism distinguishing all the eight putative species from each other by primer OPP-6 (5’- GTG GGT TGA C- 3’) and a custom primer (5’-CGC ACC GCA G-3’). RAPDprofiles generated using aforesaid arbitrary primers can serve as useful genetic markers for species identification and to improve morel systematics. Present study is the first report on exploitation of random primers in species differentiation in morels.     

Phylogenetic diversity of true morels (Morchella), the main edible non-timber product from native Patagonian forests of Argentina

Morchella species are edible fungi in high demand and therefore command high prices in world markets. Phenotypic-based identification at the species-level remains inadequate because of their complex life cycles, minor differences and plasticity of morphological characteristics between species, and the lack of agreement between scientific and common names. In Patagonia–Argentina, morels are associated with native forests of Austrocedrus chilensis (Cordilleran or Chilean cypress) and Nothofagus antarctica (ñire) and several exotic conifers that were introduced from western North America. Little is known about their taxonomy and phylogenetic relationships with other species in the genus. This work focused on the identification of collections of Morchella from Patagonia and their phylogenetic relationships with other species from the Northern Hemisphere. The comparison was made by analysis of DNA sequences obtained from four loci: the nuclear ribosomal internal transcribed spacer region (ITS) and the partial RNA polymerase I gene (RPB1) for the complete collection; and ITS, RPB1, RNA polymerase II gene (RPB2), and translation elongation factor (EF1-α) for the species-rich Elata Subclade. Analyses of individual and combined data sets revealed that Patagonian morels belong to the Elata Clade and comprised three strongly supported species-level lineages from both Patagonian native forest, and exotic trees introduced from western North America. One lineage was identified as Morchella frustrata phylogenetic species Mel-2, which is known from the USA and Canada. The second lineage, which appeared to be ‘fire-adapted’, was identified as Morchella septimelata phylogenetic species (Mel-7), which is also known from the USA. This species was collected from burned native forests mainly composed of A. chilensis and N. antarctica but also Pseudotsuga menziesii (Mirb.) Blanco, which is native to western North America. The phylogenetic analyses suggested that the third species from Patagonia was nested within the species-rich Elata Subclade and represents a new species-level lineage (informally designated Mel-37) within Elata Clade. The present collections from Patagonia constitute the southernmost latitude from which Morchella has been reported to date. The identification of two Argentine morels as North American taxa is therefore a remarkable biogeographic pattern. In view of the hypothesis that the Elata Clade originated in western North America, we speculate that at least two of the lineages colonized South America from North America via long distance dispersal, migration or, more likely, they were introduced with the exotic tree species that they were collected near.     

羊肚菌分子鉴定及功能基因研究进展

羊肚菌作为珍贵的食药用真菌,具有很高的经济价值.就近年来羊肚菌分子生物学研究的进展从分子鉴定、系统学研究、功能基因等方面进行了分析总结.结果显示,目前利用分子标记已实现羊肚菌属快捷的分子鉴定,较全面揭示了属内种群的遗传多样性和亲缘关系;利用组学技术初步探索了羊肚菌菌核形成及子实体发育机制;此外还克隆表达了与羊肚菌抗逆响...