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.     

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.     

Artificial cultivation of true morels: current state,issues and perspectives

Morels (Morchella, Ascomycota), which are some of the most highly prized edible and medicinal mushrooms, are of great economic and scientific value. Morel cultivation has been a research focus worldwide for more than 100 years, and the outdoor cultivation of morels has succeeded and expanded to a large scale in China in recent years. In this study, we review the progress in recent research regarding the life cycle and reproductive systems in the genus Morchella and the current state of outdoor cultivation. Sclerotia formation and conidia production are two important phases during the life cycle. The morel species cultivated commercially in America is M. rufobrunnea based on molecular phylogenetic analysis. The species currently cultivated in China are black morels, including M. importuna, M. sextalata and M. eximia. The field cultivation of morels expanded in the majority of the provinces in China with a yield of fresh morels of 0–7620?kg per ha. The key techniques include spawn production, land preparation and spawning, the addition of exogenous nutrition, fruiting management and harvesting. The application of exogenous nutrition is the most important breakthrough in the field of morel cultivation, but the mechanism remains unclear. It was estimated that the total amount of field cultivated fresh morels was ～500 t in 2015–2016. We also discuss the potential issues remaining in the current literature and suggest directions for future studies.     

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.     

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.     

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.     

945. RHODODENDRON BARBATUM

Rhododendron barbatum Wall. ex G. Don is a Himalayan member of Subsection Barbata (Tagg) Sleumer and the history of its discovery, subsequent introduction, habitat, ecology, cultivation and affinities are discussed.     

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.     

Assessment of local management practices on the population ecology of some medicinal plants in the coniferous forest of Northern Parts of Pakistan

A study on the assessment of local management practices on the population of three medicinal plants viz.: Persicaria amplexicaule. D. Don., Valeriana jatamansi Jones and Viola serpens Wall ex Roxb was conducted during 2002–2004 in the coniferous forest of Northern Parts of Pakistan. The objective of the study was to know the impact of current management practices on the population size of targeted plants. The study showed that the involvement of locals in the gathering of targeted plants varied with the change in elevation. Among the targeted plants V. serpens was collected by large majorities of people (83.3%) at 2700 m followed by 72% at 2300 m and 37% at 1900 m. V. jatamansi was harvested by a small number of people (18.1%) at 1900 and 2300 m each, followed by 8.3% at 2700 m. While P. amplexicaule was harvested by a few collectors (9.1%) at 1900 m and 9.6% at 2300 m followed by 8.3% at 2700 m. The study concluded that these species have been extracted so heavily in the past that they are found now sparsely in some sites of the study area. Secondly, due to loss of its habitat by deforestation and encroachment of land for cultivation its population is on the decline towards extinction. Therefore, the current study recommends the conservation of the remaining populations of targeted plants through active participation of local communities.     

William Roxburgh’s eye plant and its relevance in the nomenclature of <Emphasis Type="Italic">Phaeanthus</Emphasis> (Annonaceae)

An attempt is made to discover the identity and correct name for Uvaria tripetala Roxb., Uvaria tripetala Blanco, Phaeanthus nutans Hook. f. & Thomson and Uvaria ophthalmica Roxb. ex G. Don (Annonaceae). It is concluded that all but one belong to what has been called Phaeanthus ebracteolatus (C. Presl) Merr., which must now be referred to correctly as Phaeanthus ophthalmicus (Roxb. ex G. Don) J. Sinclair. The correct name for the species that has generally been called Phaeanthus nutans or Phaeanthus ophthalmicus is Phaeanthus intermedius (P. Parm.) I. M. Turner & Veldkamp. Typification of these names and their known synonyms is presented. This includes four new lectotypifications and the designation of two new epitypes and one new neotype.     

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.     

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.     

Smoke-water effect on the germination of Amazonian tree species

Smoke stimulates seed germination of a range of species from ecosystems that may or may not be fire prone. We evaluated the effects of smoke-water on germination of ten tree species of economic value in the Amazon region. Two materials were burnt to produce smoke-water: germination paper and the wood of Cecropia palmata Willd. Seven dilutions of the solutions were tested. Seeds of nine forest trees were germinated under controlled laboratory conditions (25 °C ± 2 °C) in the laboratory. Bertholletia excelsa Humb. & Bonpl., was tested in the nursery (approximately 25–36 °C) because of its large seeds. Irrespective of the material burned, smoke-water significantly increased seed germination of three species: Cordia goeldiana Hub., Ochroma pyramidale (Cav. ex Lam.) Urb. and Jacaranda copaia (Aubl.) D. Don. and there was a significant inhibitory effect on Swietenia macrophylla King. Germination was accelerated by smoke in J. copaia, B. excelsa and Bellucia grossularioides (L.) Triana. The most pronounced effect was observed in B. excelsa, as the mean germination time of 108 d (control) was reduced to 76 d with smoke-water made from germination paper (dilution of 1:25) and to 61 d with the one from Cecropia wood (dilution of 1:250). For five of the ten species studied, smoke-water either increased or accelerated seed germination, irrespective of the materials used for its production. Seeds with low vigour and prolonged germination time seemed to be more receptive to smoke.     

Annual dormancy cycles in buried seeds of shrub species: germination ecology of Sideritis serrata (Labiatae)

The germination ecology of Sideritis serrata was investigated in order to improve ex‐situ propagation techniques and management of their habitat. Specifically, we analysed: (i) influence of temperature, light conditions and seed age on germination patterns; (ii) phenology of germination; (iii) germinative response of buried seeds to seasonal temperature changes; (iv) temperature requirements for induction and breaking of secondary dormancy; (v) ability to form persistent soil seed banks; and (vi) seed bank dynamics. Freshly matured seeds showed conditional physiological dormancy, germinating at low and cool temperatures but not at high ones (28/14 and 32/18 °C). Germination ability increased with time of dry storage, suggesting the existence of non‐deep physiological dormancy. Under unheated shade‐house conditions, germination was concentrated in the first autumn. S. serrata seeds buried and exposed to natural seasonal temperature variations in the shade‐house, exhibited an annual conditional dormancy/non‐dormancy cycle, coming out of conditional dormancy in summer and re‐entering it in winter. Non‐dormant seeds were clearly induced into dormancy when stratified at 5 or 15/4 °C for 8 weeks. Dormant seeds, stratified at 28/14 or 32/18 °C for 16 weeks, became non‐dormant if they were subsequently incubated over a temperature range from 15/4 to 32/18 °C. S. serrata is able to form small persistent soil seed banks. The maximum seed life span in the soil was 4 years, decreasing with burial depth. This is the second report of an annual conditional dormancy/non‐dormancy cycle in seeds of shrub species.     

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.     

Conformational analysis of an anti-androgenic, (E,E)-8-hydroxygermacrene B,using NOESY and dynamic NMR spectroscopy

(E,E)-8-Hydroxygermacrene B was prepared by ketone reduction of germacrone, a naturally occurring compound from Curcuma aeruginosa Roxb. with NaBH₄ at low temperature (4 °C). This compound showed remarkable in vitro anti-androgenic activity (IC₅₀ 0.15 ± 0.022 mM) applicable to male baldness treatments. NMR analysis at −50 °C indicated that there were four conformational isomers of (E,E)-8-hydroxygermacrene B in a ratio of 48:40:8:4. The major conformers were assigned by ¹H NMR and 2D-NOESY NMR spectroscopy as having methyl groups at C-10 and C-4 in up–down (UD) orientations (48% predominance) and UU (40%). ¹H NMR spectra implied another two minor conformers with these methyls having DU (8%) and DD (4%) orientations.     

Role of calcium in alleviating effect of salinity on germination of Phragmites karka seeds

Phragmites karka (Retz.) Trin, ex. steud, a perennial reed with creeping rhizome from the family Poaceae, is distributed as pure population in brackish water swamps. Populations primarily propagate using ramets but also produce numerous seeds which form part of the seed bank after dispersal and are exposed to extremes of temperature, drought, and salinity stress. Seeds were germinated under a range of salinity (0, 100, 200, 300, 400, 500 mM NaCl) and temperature (10/20 °C, 15/25 °C, 20/30 °C, 25/35 °C, night/day) regimes in 12 h light:12 h dark photoperiod or in complete darkness with 0, 5, 10, 25 mM CaCl₂. Salinity, absence of light and high temperature (25/35 °C) reduced germination while calcium generally reversed this effect, more so at cooler temperature regimes. Calcareous soil around Karachi would help alleviate the salinity effect on the germination of P. karka and facilitate its survival.     

The varieties of Bauhinia pottsii G. Don in Thailand (Leguminosae-Caesalpinioideae)

 Bauhinia pottsii G. Don, B. subsessilis Craib, B. velutina (Wall. ex Benth.) Baker, B. mollissima (Wall.) Prain, and B. decipiens Craib were recently recognized as five varieties of Bauhinia pottsii G. Don. They grow naturally in southern Myanmar, Thailand, Indochina and the Malay Peninsula. Bauhinia pottsii var. decipiens (Craib) K. Larsen & S. S. Larsen is endemic to Thailand and known only from the type collection, so it was omitted from this study. Quantitative and qualitative morphological characters were examined in 200 specimens using multivariate and univariate analyses to determine the taxonomic relationship among the four. Some variation in qualitative characters was found between the varieties which separated them as previously defined in Flora of Thailand. Forty-three quantitative characters were subjected to cluster analysis to allow an objective classification into groups. The groups were subsequently evaluated by a canonical discriminant analysis. It was found that these characters collectively support the four varieties as defined by qualitative characters. However, the close relationship of varieties mollissima (Wall. ex Prain) K. Larsen & S. S. Larsen and velutina (Wall. ex Benth.) K. Larsen & S. S. Larsen is observed. The linear discriminant function has an overall rate of 98.5% correct classification and is useful for variety classification. Among quantitative characters, petal-claw length and ovarystalk length together with some qualitative characters are useful for key construction to separate the four varieties. Received April 24, 2001 Accepted December 11, 2001     

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.     

Seed germination ecology of the annual grass Leptochloa panicea ssp. mucronata and a comparison with L. panicoides and L. fusca

Leptochloa panicea ssp. mucronata is an annual grass that grows in relatively dry habitats. Requirements for dormancy loss and germination were determined for seeds of this species and compared to those of two species from wet habitats. Seeds of L. panicea were dormant at maturity in autumn, but when exposed to actual or simulated autumn temperatures (e.g. 20/10, 15/6 °C), they entered conditional dormancy and thus germinated to high percentages in light at 35/20 °C. Seeds buried in non-flooded soil exposed to natural seasonal temperature changes in Kentucky (USA) were non-dormant by the following summer and germinated to 80–100 % in light at 25/15, 30/15 and 35/20 °C. Seeds buried in non-flooded soil exhibited an annual conditional dormancy/non-dormancy cycle, with seeds mostly germinating to 80–100 % in light at 30/15 and 35/20 °C throughout the year but to 80–100 % in light at 25/15 °C only in summer. Results for L. panicea were compared to published data for L. panicoides and L. fusca. Whereas seeds of L. panicea buried in flooded soil failed to come out of dormancy, those of L. panicoides, an annual of moist habitats such as mudflats, exhibited an annual conditional dormancy/non-dormancy cycle, and those of L. fusca, a semi-aquatic, required flooding for both dormancy loss and germination. Differences in dormancy breaking and germination responses of seeds of Leptochloa species may help to explain why this genus occupies a wide range of habitats with regard to soil moisture conditions.