Spatial and temporal patterns of morel fruiting

The biotic and abiotic factors conditioning morel fruit body production are incompletely known. We examined spatial and temporal patterns of Morchella esculenta fruiting over five years in a wooded site in Missouri, USA. Fruiting onset was inversely correlated with spring air and soil temperatures, whereas abundance was positively correlated with rain events (>10 mm) during the 30 d preceding fruiting. The two years with the greatest fruiting had the shortest fruiting seasons (6–7 d). Fruiting season length was positively correlated with soil warming, suggesting that a narrow range of optimum soil temperatures favour the explosive production of fruit bodies. All woody stems of at least 1 cm diam were mapped and stem diameter and crown condition were noted. Morel fruit bodies were significantly closer to stems of Carya spp., Tilia americana and Ulmus americana than predicted by the frequencies of these woody species or their contribution to the total basal area on the site. Although intra-annual clustering of fruit bodies was often observed, inter-annual clustering was not. The spatial pattern of M. esculenta fruiting appears to be associated with vegetation pattern, whereas the onset and abundance of fruiting are determined by the interaction of spring temperatures with availability of supporting precipitation.     

Pattern formation: fruiting body morphogenesis in Myxococcus xanthus

When Myxococcus xanthus cells are exposed to starvation, they respond with dramatic behavioral changes. The expansive swarming behavior stops and the cells begin to aggregate into multicellular fruiting bodies. The cell-surface-associated C-signal has been identified as the signal that induces aggregation. Recently, several of the components in the C-signal transduction pathway have been identified and behavioral analyses are beginning to reveal how the C-signal modulates cell behavior. Together, these findings provide a framework for understanding how a cell-surface-associated morphogen induces pattern formation.     

The organisation of fruiting body formation in Dictyostelium minutum

The process of culmination was investigated in three strains of the species Dictyostelium minutum. After aggregates have been formed a pulsatile signalling mechanism arises; the centre of signal emission becomes the apex of the developing fruiting structure. In the late aggregate, all cells differentiate into prespore cells. Cells that have reached the apex of the culminating cells mass redifferentiate into stalk cells. In two of the three D. minutum strains, interruption of regular stalk formation, more or less random formation of stalk cells and the synthesis of stalk supporting material from cell debris often takes place. The formation of multiple apices on aggregates and early fruiting structures is characteristic for these two strains. Within the species D. minutum, the exhibition of a marked pulsatile signalling mechanism is correlated with a capacity to form a regularly shaped stalk and to organize relatively large cell masses. The possible function of pulsatile signalling in the culmination process is discussed.     

Pattern formation: fruiting body morphogenesis in Myxococcus xanthus

When Myxococcus xanthus cells are exposed to starvation, they respond with dramatic behavioral changes. The expansive swarming behavior stops and the cells begin to aggregate into multicellular fruiting bodies. The cell-surface-associated C-signal has been identified as the signal that induces aggregation. Recently, several of the components in the C-signal transduction pathway have been identified and behavioral analyses are beginning to reveal how the C-signal modulates cell behavior. Together, these findings provide a framework for understanding how a cell-surface-associated morphogen induces pattern formation.     

Beta-D-Allose inhibits fruiting body formation and sporulation in Myxococcus xanthus

Myxococcus xanthus, a gram-negative soil bacterium, responds to amino acid starvation by entering a process of multicellular development which culminates in the assembly of spore-filled fruiting bodies. Previous studies utilizing developmental inhibitors (such as methionine, lysine, or threonine) have revealed important clues about the mechanisms involved in fruiting body formation. We used Biolog phenotype microarrays to screen 384 chemicals for complete inhibition of fruiting body development in M. xanthus. Here, we report the identification of a novel inhibitor of fruiting body formation and sporulation, beta-d-allose. beta-d-Allose, a rare sugar, is a member of the aldohexose family and a C3 epimer of glucose. Our studies show that beta-d-allose does not affect cell growth, viability, agglutination, or motility. However, beta-galactosidase reporters demonstrate that genes activated between 4 and 14 h of development show significantly lower expression levels in the presence of beta-d-allose. Furthermore, inhibition of fruiting body formation occurs only when beta-d-allose is added to submerged cultures before 12 h of development. In competition studies, high concentrations of galactose and xylose antagonize the nonfruiting response to beta-d-allose, while glucose is capable of partial antagonism. Finally, a magellan-4 transposon mutagenesis screen identified glcK, a putative glucokinase gene, required for beta-d-allose-mediated inhibition of fruiting body formation. Subsequent glucokinase activity assays of the glcK mutant further supported the role of this protein in glucose phosphorylation.     

Myxococcus xanthus synthesizes a stabilized messenger RNA during fruiting body formation

Initial characterization of the unstable 5S-to-16S RNA fraction from developingMyxococcus xanthus cells reveals that it is rapidly labeled with radioactive RNA precursor and is associated with polyribosomes and released by puromycin from polyribosomes. The total unstable RNA fraction from 10-min pulse-labeled developing cells has a half-life of 13 min, compared with a 4-min half-life for unstable RNA (presumptive mRNA) from vegetative cells pulse-labeled for 2 min. We conclude that this developmental 5S-to-16S RNA contains messenger RNA and that this mRNA is stabilized compared with that in vegetative cells.     

Forest farming of shiitake mushrooms: Aspects of forced fruiting

Three outdoor shiitake (Lentinula edodes (Berk.) Pegler) cultivation experiments were established during 2002–2004 at the University of Missouri Horticulture and Agroforestry Research Center, in central Missouri. Over three complete years following a year of spawn run, we examined shiitake mushroom production in response to the temperature of forcing water, inoculum strain, substrate host species and physical orientation of the log during fruiting. Forcing compressed the period of most productive fruiting to the two years following spawn run. Further, chilled forcing water, 10–12 °C, significantly enhanced yield, particularly when ambient air temperatures were favorable for the selected mushroom strain. The temperature of water available for force-fruiting shiitake logs depends on geographic location (latitude) and source (i.e., farm pond vs. spring or well water). Prospective growers should be aware of this effect when designing their management and business plans.     

Genomics and transcriptomics to study fruiting body development: An update

Fruiting bodies of asco- and basidiomycetes are complex three-dimensional structures that protect and disperse the sexual spores. Their differentiation requires the concerted action of many genes, therefore "omics" techniques to analyze fungal genomes and gene expression at a genome-wide level provide excellent means to gain insights into this differentiation process. This review summarizes some recent examples of the use of “omics” techniques to study fruiting body morphogenesis. These include genome-centered analyses, and studies to analyze the regulation of gene expression including the analysis of RNA editing as a novel layer in the regulation of gene expression during fruiting body development in ascomycetes.     

Improved methods of isolation and purification of myxobacteria and development of fruiting body formation of two strains

By using baiting techniques and different purification methods, a high number of myxobacterial strains have been isolated as pure cultures from soil of different regions of China. Because myxobacterial cells do not disperse easily in liquid media, a medium containing an enzymatic hydrolysate of casein (CEH) medium have been used for purification and purity tests combined in a single step. The key method, in which isolates are reintroduced to sterile rabbit dung to induce fruiting bodies formation, facilitates purification of myxobacteria. Sterile rabbit dung pellets are used to mimic the natural growth substance of these organisms which has the advantage that characteristic fruiting bodies emerge, which is a key characteristics in the taxonomy of myxobacteria. In this study, the optimum program of isolation and purification of some myxobacteria strains has been established which will facilitate screening programs. Moreover, the development of fruiting body formation of strain BD20 (Chondromyces) and strain BD54 (Cystobacter) have been recorded in this study.     

Spatial organization of Myxococcus xanthus during fruiting body formation

Microcinematography was used to examine fruiting body development of Myxococcus xanthus. Wild-type cells progress through three distinct phases: a quiescent phase with some motility but little aggregation (0 to 8 h), a period of vigorous motility leading to raised fruiting bodies (8 to 16 h), and a period of maturation during which sporulation is initiated (16 to 48 h). Fruiting bodies are extended vertically in a series of tiers, each involving the addition of a cell monolayer on top of the uppermost layer. A pilA (MXAN_5783) mutant produced less extracellular matrix material and thus allowed closer examination of tiered aggregate formation. A csgA (MXAN_1294) mutant exhibited no quiescent phase, aberrant aggregation in phase 2, and disintegration of the fruiting bodies in the third phase.     

Intracellular levels of cyclic AMP and adenylate cyclase activity during mycelial development in fruiting body formation in Lentinus edodes

Abstract Streptococcal promoters were shown to fulfil primary structure requirements for the expression of heterologous genes in Pseudomonas putida KT2440. The identity of the gene products, streptokinase and human interferon-α1, as synthesized by KT2440, was confirmed by their biological, antigenic and/or physical properties. Although designed as secretion vectors with streptococcal signal sequences, the recombinant plasmids failed to mediate the complete export of either protein into the periplasmic space of KT2440.     

Role of cell cohesion in Myxococcus xanthus fruiting body formation.

Dsp mutants of Myxococcus xanthus have a complex phenotype with abnormal cell cohesion, social motility, and development. All three defects are the result of a single mutation in the dsp locus, a region of DNA about 14 kilobases long. Cohesion appears to play a central role in social motility, since nonsocial mutants exhibit weak agglutination or, in the case of Dsp cells, no agglutination (L. J. Shimkets, J. Bacteriol. 166:837-841, 1986). However, Dsp cells can be agglutinated by cohesive strains of M. xanthus. This provided the opportunity to examine the role of cohesion during development by comparing the developmental phenotype of Dsp cells with that of Dsp cells mixed with cohesive strains. Dsp mutants were unable to complete any of the developmental behaviors: aggregation, fruiting body formation, developmental autolysis, and sporulation. Contact with cohesive strains seemed to restore some developmental characteristics to the Dsp cells. When allowed to develop with wild-type cells, Dsp cells accumulated in fruiting bodies and underwent developmental autolysis, but did not form a significant portion of the spore population. Igl mutants, which may be similar to the previously described frizzy mutants, are cohesive strains that are unable to form fruiting bodies. Mixing Igl cells with Dsp cells under developmental conditions resulted in fruiting body formation, although the Dsp cells were unable to form significant levels of myxospores. In spite of their inability to sporulate under developmental conditions, Dsp mutants did not appear to be defective in the sporulation process. In fact, they formed normal levels of myxospores in response to the chemical inducer glycerol.     

Scanning electron microscopy of fruiting body formation by myxobacteria.

Scanning electron microscopy was used to follow fruiting body formation by pure cultures of Chondromyces crocatus M38 and Stigmatella aurantica. Vegetative cells were grown on SP agar and then transferred to Bonner salts agar for fructification. Fruiting in both species commences with the formation of aggregation centers which resemble a fried egg in appearance. In Chondromyces the elevated center or "yolk" region of the aggregation enlarges into a bulbous structure under which the stalk forms and lengthens. At maximum stalk height the bulb extends laterally as bud-like swellings appear. These are immature sporangia and are arranged in a distintive radial pattern around the top of the stalk. This symmetry is lost as more sporangia are formed. Stigmatella does not form a bulb; rather the yolk region of the aggregation center projects upward to form a column-like stalk which is nearly uniform in diameter throughout its length. At maximum stalk height, the terminus of the stalk develops an irregular pattern of bud-like swellings. These differentiate into sporangia. Stalks of 2-week-old mature fruiting bodies of both species appear to be cellular in composition. Stereomicrographs suggest orientation of these cells parallel to the long axis of the stalk. Stalks of 8-week-old fruiting bodies of Chondromyces were acellular and consisted of empty tubules, suggesting that the cells undergo degeneration with aging of the fruiting body.     

Heat shock protein 9 mRNA expression increases during fruiting body differentiation in Grifola frondosa and other edible mushrooms

We isolated a homolog of Schizosaccharomyces pombe HSP9 from Grifola frondosa and designated it Gf.HSP9. The Gf.HSP9 gene consisted of four exons and three introns and encoded 84 amino acid residues. We have also identified related HSP9 genes from Pleurotus eryngii (Pe.HSP9), Hypsizygus marmoreus (Hm.HSP9), and Lentinula edodes (Le.HSP9). The predicted tertiary structures of these HSP9s were the same. In addition, the expression pattern of their mRNA increased during fruiting body in a differentiation-dependent manner. These results suggested that HSP9 was associated with the fruiting body differentiation in these four edible mushrooms.     

Ribonucleic acid synthesis during fruiting body formation in Myxococcus xanthus.

A method has been devised that allowed us, for the first time, to pulse-label M. xanthus cells with precursors for ribonucleic acid biosynthesis while they were undergoing fruiting body formation. Using this method, we examined patterns of ribonucleic acid (RNA) accumulation throughout the process of fruiting body formation. As development proceeded, the rate of RNA accumulation increased at two periods of the developmental cycle: once just before aggregation and once late in the cycle, when sporulation was essentially completed. In contrast to vegetatively growing cells, in which only stable RNA species are labeled during a 30-min pulse, the majority of radioactivity found in RNA from 30-min pulse-labeled developing cells was found in an unstable heterodisperse fraction that migrated to the 5S to 16S region of sucrose density gradients and sodium dodecyl sulfate-polyacrylamide gels. This pattern of incorporation could not be induced (i) by a shift down of vegetatively growing cells to a nutritionally poor medium, in which the generation time was increased to that of developing cells during the growth phase, or (ii) by plating of vegetative cells onto the same solid-surface environment as that of developing cells, but which surface supported vegetative growth rather than fruiting body formation. Thus, the RNA synthesis pattern observed appeared to be related to development per se rather than to nutritional depletion or growth on a solid surface alone. The radioactivity incorporated into the unstable 5S to 16S RNA fraction accumulated as the pulse length was increased from 10 to 30 min; in contrast, an analogous unstable fraction from vegetative cells decreased as pulse length was increased. This suggested that developmental 5S to 16S RNA was more stable than vegetative cell 5S to 16S RNA (presumptive messenger RNA). However, during a 45-min chase period, radioactivity in 30-min-pulse-labeled developmental 5S to 16S RNA decayed to an extent twice that of developmental RNA located in 16S and 23S regions of sucrose density gradients and was considerably less stable than the 5S, 16S, and 23S RNA species labeled during a 30-min pulse of vegetative cells.