Internuclear Genetic Transfer in Vegetative Dikaryons of SCHIZOPHYLLUM COMMUNE: I. Di-Mon Mating Analysis

A series of hemi-compatible dikaryon x monokaryon (di-mon) matings was designed to determine whether there was any genetic interaction between the dikaryotic nuclei. One of the nuclei in each dikaryon was known to carry a recessive gene (mnd) that promoted the development of an abnormal growth form, mound. Dikaryons containing both mnd ⁺ and mnd nuclei produced mosaic colonies that consisted of three distinct kinds of hyphae: mound, fruiting body, and vegetative (devoid of any multihyphal structure). When dikaryotic hyphae from each of these morphological regions were used in di-mon matings, the genetic and developmental characteristics of the selected nuclear types were examined in the resulting derived dikaryons. The results showed that fruiting-body and vegetative cells contained the expected mnd and mnd⁺ nuclei. Dikaryotic mound hyphae, however, contained only mnd nuclei. In a manner as yet unresolved, but clearly dependent on the presence of the mnd allele, the mnd ⁺ allele of a wild nucleus was altered to, or acquired, the mnd allele. A number of hypotheses were considered to explain the genetic event(s) that generates [mnd + mnd*] dikaryotic cells from [mnd⁺ + mnd] cells, but none was found to be entirely satisfactory.     

Genetic Control of Recombination in SCHIZOPHYLLUM COMMUNE: Location of a Gene Controlling B-Factor Recombination

In S. commune the frequency of recombination between the two subunits, α and β, of the B incompatibility factor is genetically controlled. Analysis of the progeny obtained from crosses between high- and low-recombining strains indicates that the gene controlling recombination frequency in the B factor is linked to the B factor itself, approximately nine map units from Bβ. This gene, called B-rec-1, does not affect the recombination frequency in an unlinked region (between the subunits of the A incompatibility factor) or in a region contiguous with the B factor (between Bα and the morphological marker dome-2).     

POLYOLS IN SCHIZOPHYLLUM COMMUNE

Sugars and sugar alcohols present in extracts of the wood-rotting mushroom Schizophyllum commune were identified by paper chromatography during fruiting, basidiospore germination, and growth of vegetative mycelium. Homokaryotic fruitbodies and dikaryotic fruits derived from several compatible matings of S. commune contained mannitol and arabitol. Basidiospores shed from dikaryotic fruits also contained mannitol and arabitol while the latter disappeared during spore germination. Vegetative mycelium (strain 699) contained glucose, fructose, mannitol and glycerol while these compounds as well as arabitol occurred in mycelium of strain 845. Polyols are not, therefore, associated exclusively with the sporulation process in S. commune.     

Internuclear Genetic Transfer in Dikaryons of SCHIZOPHYLLUM COMMUNE. II. Direct Recovery and Analysis of Recombinant Nuclei

The recessive gene, mound (mnd), allows the appearance of globose masses of compacted hyphae. Dikaryons of Schizophyllum commune that are heteroallelic for mnd [(mosaic dikaryons: (mnd + mnd(+))] have been successfully dedikaryotized in cholate-containing medium in order to recover the component nuclear types directly. The relative proportion of the two recovered monokaryotic types shows in all cases a marked deviation from 1:1. Hyphae from nonmound mycelial regions yield monokaryotic types identical to those originally used to form the dikaryons. In hyphae from mound-forming regions, however, homoallelism of the mnd allele has been demonstrated; the nuclear type that formerly contained the mnd(+) allele acquired a mnd allele.-The process of internuclear transfer or recombination is unaccompanied by the simultaneous alteration of any additional genetic markers carried by the recipient nucleus. The newly acquired mnd allele segregates in Mendelian fashion in subsequent outcrosses and appears to be chromosomally located. A novel process of somatic recombination, with several features distinct from classical parasexual mitotic recombination, appears to be in operation.     

A Class of Genes Affecting B Factor-Regulated Development in SCHIZOPHYLLUM COMMUNE

Twelve mutations affecting nuclear migration, a major developmental phase in Schizophyllum commune, display a complex pattern of complementation and recombination. They are expressed only when a genetic factor controlling this phase of development, the B incompatibility factor, is operative. All twelve mutations are linked to the B factor, nine in a cluster and three in distinct loci outside the cluster. A linear map cannot be constructed from the frequency of recombination. Complementation maps are not linear. There is little correlation between the complementation groups and the groups based on recombination. Many pairs of mutations that do not complement recombine with frequencies of 1.1% to 26.9%. The genes represented by the twelve mutations are located in a linked group of about 18 known genes involved in the specific phase of development controlled by the B factor.     

NUCLEAR MIGRATION IN DIKARYOTIC-HOMOKARYOTIC MATINGS IN SCHIZOPHYLLUM COMMUNE

The present data indicate internuclear selection in dikaryotic-homokaryotic matings is not due to differential rates of migration of the 2 types of migrant nuclei or to differential rates of saturation of the homokaryon. The advancing fronts of the 2 types of migrating nuclei are separated by essentially the same distance shortly after migration has been initiated and during later periods in the migration process. Internuclear selection, controlled by the incompatibility factors, must occur during the initiation of migration or earlier. Migration of both types of nuclei from the dikaryon has been demonstrated in hemicompatible and incompatible matings.     

New Mutations and a 7-Chromosome Linkage Map of SCHIZOPHYLLUM COMMUNE

Forty-eight useful new mutations of S. commune were obtained by mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine. Their requirements and meiotic linkage relationships to each other and previously mapped areas were investigated. Several of these new mutations were incorporated into diploid strains so that the diploids contained at least one marker on every linkage group. Analysis of somatic recombination in these diploids indicated that each meiotic linkage group corresponded to an independent chromosome.     

MORPHOGENESIS OF SCHIZOPHYLLUM COMMUNE I. MORPHOLOGICAL VARIATION AND MATING BEHAVIOR OF THE THIN MUTATION

A single gene, recessive mutation of Schizophyllum commune, designated as thin, is characterized by its sparse growth on agar and unusual hyphal morphology. Many hyphae exhibit various degrees of “waviness,” although some branches of these wavy hyphae appear normal. Differences in cell size and branch development between normal and thin strains are also noted. Most of the characteristics attributed to thin vary quantitatively between strains. In addition, considerable variability in hyphal characteristics can be found in a single strain. Thin mutants behave as bilateral maters. In matings where the normal allele is present, genetic complementation occurs and normal clamped cells are produced. Fruit bodies with viable spores are formed on both the thin and normal sides of the mating. Thin X thin matings give a stable dikaryon, but they retain the thin morphology and do not produce fruit bodies. Evidence that there is only one locus for the thin mutation is presented. Certain aspects of the variability of thins, the possible role of extracellular polysaccharide, and the potential uses for thin strains in studies of hyphal and fruit body development are discussed.     

COBALT AS A MITOCHONDRIAL DENSITY MARKER IN A STUDY OF CYTOPLASMIC EXCHANGE DURING MATING OF SCHIZOPHYLLUM COMMUNE

A slow in vivo uptake of cobalt from a growth medium resulted in an increase in density of mitochondria of Schizophyllum commune. Differential labeling of donor and resident mycelia, and subsequent analysis of resident mycelia surrounding donor implants, detected cobalt-dense mitochondria and demonstrated exchange of mitochondria after hyphal fusion. Transfer of mitochondria occurred in fully compatible, common-A, and common-AB matings, but was not detected in common-B matings of the tetrapolar Basidiomycete S. commune.     

Mutational Analysis of Natural Alleles at the B Incompatibility Factor of SCHIZOPHYLLUM COMMUNE: alpha2 and beta6

The B incompatibility factor of the fungus Schizophyllum commune having allelic specificity α2–β6 was subjected to mutagenesis by X-irradiation. Five types of mutations were recovered, four of them new types previously unreported. All lead to loss of the B-factor regulatory function; three of the five have retained their allelic specificity. The mutations map in three closely linked sites: Bα, Bβ, and between Bα and Bβ.     

Dependence of Vegetative Recombination Among Haemophilus influenzae Bacteriophage on the Host Cell

Vegetative recombination of temperature-sensitive mutants of Haemophilus influenzae phage HP1 cl was measured in wild-type H. influenzae strain Rd and in strain DB117, an ultraviolet-sensitive, transformation-defective mutant of the Rd strain. Recombinants are formed with low frequency in wild-type cells, but no recombination was detectable in DB117. It is concluded that these phage make use of the host cell enzymes for vegetative recombination. Lysogenization readily takes place in both strains.