Culture and reproductive systems of 11 species of Mycetozoans

The culture and reproductive systems of 10 species (16 isolates) of myxomycetes and one species (one isolate) of protostelid were investigated. A single isolate of Ceratiomyxa fructiculosa was grown on agar and found to be nonheterothallic. This is the first report of spore-to-spore cultivation of this species and the first report of a reproductive system in the protostelids. Isolates of the myxomycetes Didymium dubium, Didymium iridis, Didymium vaccinum, Licea biforis, Perichaena vermicularis, Physarum gyrosum, Physarum pusillum (six isolates) and Semimorula liquescens all were nonheterothallic. This is the first report of culture and a reproductive system for D. vaccinum, the first report of nonheterothallism for S. liquescens and the second report of nonheterothallic isolates of D. dubium, Licea biforis, Perichaena vermicularis and P. gyrosum. The nonheterothallic isolate of D. iridis is one of many reported for this species, and the six nonheterothallic isolates of P. pusillum add to the seven nonheterothallic and two heterothallic isolates already known. In addition, five of the isolates of P. pusillum apparently represent a small form that is adapted to an ephemeral micohabitat, and the sixth is a yellow form of a species that is typically white. The Didymium ?ovoideum isolate and the two Physarum didermoides isolates have heterothallic reproductive systems. The D. ?ovoideum isolate is somewhat different from most isolates of this species in its morphology and reproductive system. It is not compatible with any of the heterothallic isolates of long-stalked Didymium, including the A0 biological species already determined for D. ovoideum; therefore, it is either a new biological species of D. ovoideum or a separate new species. The two heterothallic isolates of P. didermoides form a multiple allelic mating-type series with four alleles.     

PRELIMINARY INVESTIGATION OF THE DOUBLE-DIFFUSION TECHNIQUE AS A TOOL IN DETERMINING RELATIONSHIPS AMONG SOME MYXOMYCETES,ORDER PHYSARALES

Double-diffusion technique was used to investigate myxomycete relationships within the order Physarales. Extracts of plasmodia of 22 slime mold isolates were reacted with five antisera produced to Plasmodia of Didymium nigripes, Physarella oblonga, Physarum polycephalum, Physarum gyrosum and Fuligo septica. Two isolates of Fuligo septica tested alike. Four isolates of Physarum pusillum did not test alike, and no valid conclusion of the relationship of this species was possible. These isolates showed strong serological affinity: (1) Physarum gyrosum, Physarella oblonga, two isolates of Fuligo septica, and possibly two isolates of Physarum pusillum, and Physarum tenerum; (2) Physarum polycephalum and Physarum flavicomum; (3) Fuligo septica and many of the species tested; (4) Didymium nigripes and at least one isolate of Didymium iridis. In most cases serologial relationships among species tested did not coincide with current taxonomy based on morphology of fructification.     

Nivicolous Myxomycetes from tyrol (Austria). I

25 taxa of Myxomycetes collected around melting snow banks in montaneous and alpine areas of Tyrol are presented. From a chorological point of view especially interesting are: Diderma asteroides, D. lyallii, D. peyerimhoffii, Didymium nivicolum, Hemitrichia abietina, Physarum alpestre, Trichia persimilis, Trichia sordida var. sordida, Trichia sordida var. sordidoides. The nivicolous taxa Badhamia alpina and Didymium wilczekii are considered to be synonyms of Badhamia foliicola and Didymium dubium. SEM micrographs of spores and capillitia of the most significant species are included.     

HETEROTHALLISM AND HOMOTHALLISM: A STUDY OF 27 ISOLATES OF DIDYMIUM IRIDIS,A TRUE SLIME MOLD

Single pore studies of 27 isolates of Didymium iridis from several geographical locations revealed that 11 are heterothallic, displaying a one-locus, multiple allelic mating system. Collectively, 9 of the 11 possess a total of 12 mating types in one multiple allelic series. Each of the remaining two isolates has a pair of mating types which are either incompatible with the 12 or they have not yet been tested against them. Sixteen isolates proved to be homothallic.     

Distribution and sporulation phenology of myxomycetes in the Sonoran Desert of Arizona

All pith samples from 68 dead saguaro cacti in 3 plots and 11 isolated dead plants in Saguaro National Monument, Arizona, produced at least one species of myxomycete upon incubation at 20 or 30°C. Three species,Badhamia gracilis (Macbr.) Macbr.,Physarum straminipes Lister, andDidymium eremophilum M. Blackwell et Gilbertson, developed at high frequencies on the substrates in moist chamber culture.Perichaena corticalis (Batsch) Rost, andProtophysarum phloiogenum M. Blackwell et Alexopoulos were also present. Although previous literature reports [9] indicated that Myxomycetes grow best at low pH, these species all tolerated substrates of pH 8.7–10.4.Didymium eremophilum andP. phloiogenum had peaks in sporulation within 6 days; other species were slower. There was no difference in time of sporulation ofB. gracilis orD. eremophilum at 20 and 30°C; however, sporulation ofP. straminipes was significantly later at 30°C. Reduced spore germination and slower buildup of critically sized amoebal populations ofP. straminipes at 30°C may be a factor.     

Myxomycetes from Orissa (India)-III

Summary Six myxomycetous fungi,Cribraria intricata Schrad.,Elaeomyxa miyazakiensis (Emoto)Hagelst.,Stemonitis flavogenita Jahn,Didymium vaccinum (Dur. &Mont.)Buchet,Didymium minus (Lister)Morgan andPhysarella oblonga (Berk. &Curt.)Morgan have been figured and described form Cuttack, Orissa, of whichElaeomyxa miyazakiensis (Emoto) Hagelst.,Stemonitis flavogenita Jahn,Didymium vaccinum (Dur. &Mont.)Buchet,Didymium minus (Lister)Morgan are new records for India.     

CYTOPHOTOMETRIC MEASUREMENT OF NUCLEAR DNA IN SEVEN HETEROTHALLIC ISOLATES OF DIDYMIUM IRIDIS,A MYXOMYCETE

Absorption cytophotometry was used to measure nuclear Feulgen-DNA content of myxamoebae and Plasmodia in seven heterothallic isolates of Didymium iridis. Measurements of myxamoebal nuclei from clones of four isolates (Hon 1, Pan 1, Pan 2, and CR 5) gave a mean DNA value of 0.34, whereas the nuclei of Plasmodia which develop from each of the four intraisolate crosses had a mean value of 0.63. These values correspond to the 2C haploid level in myxamoebae and the 4C diploid level in Plasmodia. DNA values in two additional isolates (Pan 3 and CR 2) are much higher than the mean for the other five. Accordingly, it is proposed that these may be polyploid. The question of polyploidy in D. iridis and in other myxomycetes is evaluated. The seventh isolate, Ky 1, is taxonomically very close to D. nigripes and was not included in calculations of mean values for D. iridis.     

NUCLEAR DNA CONTENT OF MYXAMOEBAE AND PLASMODIA IN SIX NON-HETEROTHALLIC ISOLATES OF A MYXOMYCETE,DIDYMIUM IRIDIS

The nuclear DNA content of six non-heterothallic isolates of the myxomycete Didymium iridis was measured by combining the Feulgen reaction with absorption microspectrophotometry. This allowed us to distinguish between homothallic (sexual) and apogamic (non-sexual) isolates. Four of the isolates studied, Panamanian 4 and 5, California 1, and Missouri 1 are homothallic. Moreover, the average DNA content of the myxamoebal and plasmodial nuclei (0.32 and 0.61 respectively) does not differ significantly from the calculated haploid and diploid values for heterothallic isolates of D. iridis (0.34 and 0.63). Hence, it is concluded that in each of these isolates the myxamoebae are haploid and the plasmodia diploid. In two of the isolates investigated, Georgia 1 and Hawaii 1, the DNA content of the myxamoebal and plasmodial nuclei did not differ significantly. Therefore, in both of these isolates the plasmodia appear to develop apogamically. In addition the mean DNA values recorded for the Ha-1 isolate suggest that it is aneuploid.     

COMPARATIVE MEASUREMENTS OF NUCLEAR DNA IN A HETEROTHALLIC AND A SELF-FERTILE ISOLATE OF THE MYXOMYCETE,DIDYMIUM IRIDIS

Comparative measurements were made of the nuclear Feulgen-DNA content of a heterothallic and a self-fertile isolate of the myxomycete Didymium iridis. Plasmodial nuclei of both isolates contain the diploid amount of DNA. The replicated diploid (4C) values for the heterothallic and the self-fertile isolates are 5.66 and 5.95, respectively. Myxamoebae, however, are quite dissimilar in their nuclear DNA content. Those of the heterothallic isolates, Honduran 1–2 (A¹) and Panamanian 2–4 (A⁷), have mean values of 3.81 and 3.69, whereas myxamoebae of the self-fertile Philippine-1 isolate were found to have a mean value of 6.07. Myxamoebae of the Ph-1 isolate are, therefore, at the same ploidy level as the Ph-1 Plasmodium. Mean DNA values for Ph-1 sporangial nuclei were in category 4C. Measurement of the DNA content of mitotic metaphases in sporangia at T = 6 hr confirmed that the mean DNA content of both Ph-1 myxamoebae and plasmodial nuclei is equivalent to 4C. It is concluded that nuclear phase alternance is lacking in the Ph-1 isolate and that the Plasmodium of this isolate develops by apogamy.     

Macrocyst formation in three dictyostelid species,Dictyostelium monochasioides, Polysphondylium candidum, andP. pseudo-candidum

Macrocyst formation in the sexual cycle was found in three dictyostelid species:Dictyostelium monochasioides, Polysphondylium candidum, andP. pseudo-candidum. Mating tests suggested thatD. monochasioides andP. pseudocandidum were heterothallic andP. candidum was homothallic. The primary walls of macrocysts had partially or fully degenerated, while the inner walls, believed to be tertiary walls, showed an undulate structure.     

Mortierella sugadairana,a new homothallic species related to the firstly described heterothallic species in the genus

A new species, Mortierella sugadairana, is described for a fungus forming homothallic zygospores with a club-shaped macrosuspensor and a microsuspensor originating from the macrosuspensor. The species was isolated from cool regions in Japan and morphologically and phylogenetically close to a heterothallic species M. parvispora, which is the first species reported as a heterothallic species in the genus. Mycelial growth of the species was limited at 30 °C, whereas two isolates of M. parvispora can grow. This may indicate that the species and M. parvispora adapted to different climates from a common ancestor involving differentiation of the manner of reproduction.     

INTER-THALLIC SEXUAL REACTIONS IN ACHLYA,A GENUS OF THE AQUATIC FUNGI

Barksdale , Alma Whiffen . (New York Botanical Garden, New York.) Inter-thallic sexual reactions in Achlya, a genus of the aquatic fungi . Amer. Jour. Bot. 47(1): 14—23. Illus. 1960.– Inter-thallic sexual reactions, involving the coöperative induction of sexual organs, have been found to occur between members of homothallic, as well as heterothallic species of the sub-genus Achlya. The induction of sexual organs, both on single thalli and on paired thalli, is temperature-dependent. Sexual organs were initiated at 15°—20°C. in 14 of 27 heterothallic strains that were usually sterile at 25°. The hermaphroditic sexual organs were like those of either A. ambisexualis or A. bisexualis in 19 strains; like those of A. prolifera in 3 strains; like those of A. inflata in 1 strain; and like those of A. americana in 1 strain. In some instances, the sexual reaction between paired thalli was complete at 20°C. but incomplete (i.e., oospores were not formed) at 25°C. The heterothallic and homothallic strains studied could be assigned to 1 of 12 (6 female and 6 male) inter-action types, on the basis of their sexual responses. These sexual types are characterized and their possible usefulness in indicating kinship among species of Achlya is assessed. The possible origin of heterothallic from homothallic forms of Achlya is discussed.     

Pigmentation and sporulation in selected Myxomycetes

Chemical, chromatographic and spectrometric methods are used to characterize plasmodial pigments and determine relationships between pigmentation and sporulation in selected Myxomycetes. In Physarum gyrosum (white) a single pigment is identified and characterized as a flavone. Physarum polycephalum (yellow) and Didymium iridis (brown) contain four and six components, respectively, in their plasmodial pigments which test negatively for flavones but show the presence of some type of phenolic compound. No detectable component is identified in the white plasmodium of Didymium squamulosum which proved to be independent of light for fruiting. The absorption spectra of all species that were light sensitive for fruiting showed common peaks in the 300–400-mμ region, among others. Pigment changes associated with light absorption are reported for some white, yellow and brown plasmodial types. In Physarum gyrosum a yellow pigment forms in light which did not show the characteristic flavones present in the white plasmodial stage. Changes in absorption spectra are reported for Physarum polycephalum, Didymium iridis and Didymium squamulosum as the plasmodial pigments change prior to fruiting. Results show a close relationship between the physiology of plasmodial pigmentation and sporulation in the Myxomycete species studied.     

青海黏菌的研究(英文)

通过对2009年采集于青海省互助县北山、循化县孟达和大通县鹞子沟3个国家自然保护区或森林公园的65份黏菌标本的鉴定和对中国科学院微生物研究所真菌标本馆保藏的来自青海乐都县、民和县、湟源县、祁连县、乌兰县、门源县和西宁市的29份黏菌标本的复核,共明确青海已知黏菌5目8科22属36种1变种,其中,格斑双皮菌Diderma roanense为中国新记录种,另有19种黏菌为青海省黏菌新记录种,为中国青藏高原及高海拔山地分布的黏菌补充了新的资料。     

DNA homologies among homothallic, pseudo-homothallic and heterothallic species of Neurospora.

Summary Highly purified DNAs from three homothallic speciesNeurospora africana, N. dodgei andN. lineolata; three reference strains representing authentic heterothallic species,N. crassa, N. intermedia andN. sitophila; and two strains of pseudo-homothallic speciesN. tetrasperma were characterized by spectrophotometry and DNA reassociation using hydroxyapatite chromatography. All of these known species are closely related on the basis of DNA characteristics such as base composition and thermal denaturation profiles of major DNA components. Minor components of ascospore DNA was, however, only 5–7% of total DNA instead of 15–20% minor component DNA shown by mycelial DNA. Species belonging to same group were not distinguishable morphologically, but all of these species were distinguishable by DNA:DNA homology studies. Greater DNA homology was noticed between DNAs of heterothallic species and DNAs of pseudohomothallic species than DNA of true homothallic species. Difference on DNA-nucleiotide sequences among homothallic species was very little. Pseudo-homothallic speciesN. tetrasperma was found to be distinctly different from homothallic species but closer to heterothallic species based on such studies.Supported in part by a contract No. E(40-1)4182 with the U.S. Energy Research and Development Administration. We are grateful to Departments of Oncology and Radiotherapy, College of Medicine, Howard University, Washington, D.C. for providing us with material assistance     

Concerted evolution of a tandemly arrayed family of mating-specific genes in Phytophthora analyzed through inter- and intraspecific comparisons

Multigene families are features of most eukaryotic genomes, which evolve through a variety of mechanisms. This study describes the structure, expression, and evolution of a novel family in the oomycete Phytophthora. In the heterothallic species P. infestans, M96 is expressed specifically during sexual sporogenesis, and encodes a low-complexity extracellular protein that may be a component of oospore walls. Intriguingly, M96 exists in P. infestans as 22 relatively homogeneous loci tandemly repeated at a single site, which is partitioned by inversions and retroelements into subclusters exhibiting semi-independent evolution. M96 relatives were detected in other heterothallic and homothallic oomycetes including species closely (P. mirabilis, P. phaseoli) or distantly (P. ramorum, P. sojae) related to P. infestans. Those M96 relatives also exhibit oosporogenesis-specific expression and are arrayed multigene families. Nucleotide changes and repeat expansion diversify M96 in each species, however, paralogues are more related than orthologues. Concerted evolution through gene conversion and not strong purifying selection appears to be the major contributor to intraspecific homogenization. Divergence and concerted evolution was also detected between isolates of P. infestans. The divergence of M96 proteins between P. infestans, P. ramorum, and P. sojae exceeds that of typical proteins, reflecting trends in reproductive proteins from other kingdoms. Sequence data from this article have been deposited with the GenBank Data Libraries under accessions DQ196155 to DQ196175.     

Heterokaryon formation in Thanatephorus cucumeris anastomosis group 2-2 IV

Thirty single basidiospore isolates (SBIs) obtained from four field isolates of the basidiomycete fungus Thanatephorus cucumeris AG 2-2 IV were examined for heterokaryon formation. SBIs of three of four field isolates (Rh509, 92155 and R94) did not produce a tuft of mycelium in the hyphal interaction zone between paired isolates on 2% charcoal agar. Field isolates Rh509, 92155 and R94 indicated no death of interacting mycelium with their progenies on glass slide and microscopic examination. AFLP (amplified fragment length polymorphism) phenotypes of parent and their SBIs were identical. Field isolates Rh509, 92155 and R94 and their SBIs were homothallic. SBIs obtained from field isolate SA-1 were grouped into two mating types (SBI-M1 and SBI-M2), and a tuft of mycelium was formed between paired SBIs-M1 and -M2. SBIs of field isolate SA-1 indicated that no death and death of interacting mycelium were randomly observed. AFLP phenotypes among SBIs of isolate SA-1 were not identical and were also different from their parent isolate. AFLP phenotypes of tuft mycelia produced between heterothallic SBI-M1 and -M2 were heterokaryotic. The mating system of field isolate SA-1 and its SBIs was heterothallic. Both SBIs-M1 and -M2 further produced tuft mycelium with homothallic field isolates and their SBIs. AFLP banding patterns suggested that tuft mycelium was heterokaryotic produced from between heterothallic and homothallic isolates. Results from these experiments clarified that both homothallic and heterothallic isolates exist in population of T. cucumeris AG 2-2 IV, and that genetic exchange can occur between homothallic and heterothallic isolates.     

Synaptonemal complexes and meiosis in myxomycetes

Synaptonemal complexes (SC) have been observed in spores 18–24 hr past cleavage in natural fruitings of Physarum cinereum, P. bogoriense, Hemitrichia stipitata, Tubifera ferruginosa, and Arcyria incarnata. Laboratory fruitings of Arcyria cinerea, Stemonitis herbatica, and a homothallic isolate of Physarum pusillum also have SC's present in spores during the same postcleavage period. The presence of these paired chromosomes of meiotic prophase in spores of species collected in nature and in a diversity of taxa suggests that the usual position of meiosis in Myxomycetes is inside the postcleavage spore. Criteria are proposed for evaluating the validity of the SC as an indicator of meiosis.     

Bacterial Endosymbionts of Pyrodinium bahamense var. compressum

The study presents evidence in support of the bacterial theory associated with the toxicity of Pyrodinium bahamense var. compressum. Bacterial endosymbionts from Philippine P. bahamense var. compressum strain Pbc MZRVA 042595 were isolated and identified via 16S rDNA sequence analysis. Taxonomic diversity of the identified culturable intracellular microbiota associated with Philippine P. bahamense var. compressum was established to be limited to the Phyla Proteobacteria, Actinobacteria, and Firmicutes. Major endosymbionts identified included Moraxella spp., Erythrobacter spp., and Bacillus spp., whereas Pseudomonas putida, Micrococcus spp., and Dietzia maris were identified as minor isolates. All identified strains except D. maris, P. putida, and Micrococcus spp. were shown to contain either saxitoxin or neo saxitoxin or both at levels ≤73 ng/10⁷ bacterial cells based on high-performance liquid chromatography analysis. Paralytic shellfish poisoning-like physiologic reactions in test animals used in the mouse assay were recorded for the endosymbionts except for P. putida. The study is the first to elucidate the possible contribution of bacterial endosymbionts in the toxicity of P. bahamense var. compressum isolated in the Philippines.