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.     

Role of a cell-wall glucan-degrading enzyme in mating of Schizophyllum commune

Mycelial enzyme extracts of Schizophyllum commune were prepared during vegetative growth matings leading to common-A and common-B heterokaryons and the dikaryon, and were examined for hydrolytic activity against an alkaliinsoluble cell-wall glucan (R-glucan) isolated from this mushroom. In extracts from several individual homokaryotic mycelia the R-glucanase activity was low and did not increase when the cultures exhausted glucose in the medium. In common-A matings, a 30-fold increase in specific activity of intracellular R-glucanase was found even in the presence of glucose in the broth. An increase of this magnitude was not observed in the common-B mating nor in the fully compatible cross leading to the dikaryon. Extracts of the dikaryon did show elevated R-glucanase activity after exogenous glucose disappearance and subsequent fruiting. In none of these situations was an enzyme activity detected towards an alkali-soluble cell-wall glucan (S-glucan) prepared from S. commune. Changes in R-glucanase were not parallelled by identical changes in laminarinase, pustulanase, cellobiase, and p-nitrophenyl-beta-d-glucosidase, but comparable increases in specific activities were found for hydrolysis of glycogen and maltose. After interaction of the various mycelia in mating combinations, the S-glucan/R-glucan ratio of the cell wall of the dikaryon was found to be similar to that of the homokaryons, but increased in the common-B interaction and was elevated almost threefold in the common-A heterokaryon.     

Abo blood groups and completed reproductive performance of rural Haryanavi couples: analysing measures of selection intensities

The possible differential effects of ABO blood group materno-paternal (fetal) incompatibility on completed reproductive performance were investigated on a sample of 100 couples (100 fathers and 100 mothers) from three villages in the Jind district of Haryana state, India. The average number of live births per mating couple was slightly higher for the incompatible matings (5.32) than the compatible ones (5.05). This advantage was offset by higher postnatal mortality in the former. Consequently, the average number of living children in the compatible matings (4.64) was higher than in the incompatible ones (4.18). With reference to individual ABO matings, the index of relative fertility (Irf) was the highest in A x AB followed by B x A type of incompatible matings. No decrease in live births in O x A and O x B incompatible matings was observed compared with their reciprocal compatible ones, i.e. A x O and B x O matings, as has been hypothesized in previous studies. The total pregnancy wastage was substantially higher in ABO-incompatible matings (24.59%) than compatible matings (8.45%). About 71% of the postnatal deaths took place within one year of the birth in the case of incompatible matings compared with 50% in the case of compatible matings. The study supports the hypothesis that selection is operative at the ABO locus as revealed by the measures of selection intensity. The loss of fitness in the present sample was associated with differential mortality. There were no differences in the proportions of average number of male live births in the compatible (0.55) and incompatible matings (0.58). However, in the individual mating types, there was some evidence of higher or lower proportions of male live births.     

THE KINETICS OF INITIAL NUCLEAR EXCHANGE IN COMPATIBLE AND NONCOMPATIBLE MATINGS OF SCHIZOPHYLLUM COMMUNE

The effect of mating-type factors on exchange of nuclei between mycelial fragments mated in liquid culture was studied. Evidence for exchange of nuclei was based on three criteria for detecting nuclei from both mating partners in mycelial fragments of the developing dikaryon or hetero-karyon. The kinetics of nuclear exchange were shown to be relatively independent of the method used. It was shown that the kinetics of nuclear exchange in the first 96 hr are different in the compatible (A41 B41 × A42 B42), common-A (A42 B42 × A42 B41), common-B (A41 B42 × A42 B42), and common-AB (A41 B41 × A41 B41) matings. In all four types of matings, the percentage of fragments possessing both types of nuclei 12-24 hr after mating is nearly equal. After this time, significant differences appear in the patterns for compatible vs. the three non-compatible matings and the common-A vs. the common-B and common-AB matings. The percentage of mycelial fragments possessing both types of nuclei throughout the 96-hr test period is similar for both the common-B and common-AB matings. The kinetics of nuclear exchange were shown to be independent of the particular mating-type alleles and nutritional markers used. When the efficiency of nuclear exchange in complete and minimal media was compared, it was shown that nuclear exchange occurred more rapidly and synchronously in minimal medium. This difference is not due to growth differences in the two media. These data indicate that the earliest mating interactions, i.e., hyphal anastomosis and nuclear exchange, are independent of the mating-type factors but that subsequent events are determined by these genes.     

Regulation of the pattern of protein synthesis in Schizophyllum commune by the incompatibility genes

The pattern of protein synthesis in various coisogenic mycelial types of Schizophyllum commune, viz. monokaryon, dikaryon, and homokaryons carrying primary mutations in the A and the B factors, was studied by two-dimensional gel electrophoresis. After pulse-labeling with ³⁵S-methionine, approximately 650 of 710 proteins analyzed were common to all mycelial types. Coisogenic monokaryons differed by only 2%, whereas the largest difference was found between these monokaryons and the dikaryon derived from them (6.6 and 7.7%). The majority of these differences fell into two about equally sized categories, i.e., proteins which were either specifically absent (“switched-off” proteins) or present (“switched-on” proteins) in the dikaryon. “Switched-on” proteins were on the average larger and slightly more acidic than “switched-off” proteins. The double factor mutant which best mimicked the dikaryon in morphology also best resembled the dikaryon in types of proteins synthesized. Unexpected, however, was the large overlap in proteins apparently controlled by each of the two incompatibility factors individually, despite the distinct morphological sequences directed by each of them.     

Sexual selection in mushroom-forming basidiomycetes

We expect that sexual selection may play an important role in the evolution of mushroom-forming basidiomycete fungi. Although these fungi do not have separate sexes, they do play female and male roles: the acceptance and the donation of a nucleus, respectively. The primary mycelium (monokaryon) of basidiomycete fungi, growing from a germinating sexual spore, is hermaphroditic, but it loses female function upon the acceptance of a second nucleus. The resulting dikaryon with two different nuclei in each cell retains a male potential as both nuclei can fertilize receptive mycelia. We tested the occurrence of sexual selection in the model species of mushroom-forming basidiomycetes, Schizophyllum commune, by pairing monokaryons with fully compatible dikaryons. In most pairings, we found a strong bias for one of the two nuclei although both were compatible with the monokaryon when paired alone. This shows that sexual selection can occur in mushroom-forming basidiomycetes. Since the winning nucleus of a dikaryon occasionally varied depending on the receiving monokaryon, we infer that sexual selection can operate through choosiness of the receiving individual (analogous to female choice). However, in other cases the same nucleus won, irrespective of the receiving monokaryon, suggesting that competition between the two nuclei of the donating mycelium (analogous to male–male competition) might also play a role.     

Fungal fidelity: Nuclear divorce from a dikaryon by mating or monokaryon regeneration

Basidiomycete fungi perform fertilizations by incorporation of nuclei into a monokaryotic mycelium to establish a dikaryon. The dikaryon cannot incorporate another type of nucleus, but can still act as a nucleus donor in a dikaryon–monokaryon (di–mon) mating, known as the Buller phenomenon. Previously, it has been observed that: (1) in a particular di–mon mating, one of the nuclear types of the dikaryon generally performs better as a donor than the other, and (2) when nuclei from a dikaryon are separated to form monokaryons again (dedikaryotisation), recovery of monokaryons of the two nuclear types is usually unequal. In this study, we investigated if these two observations of asymmetry are functionally related. We tested this hypothesis by performing both di–mon matings and dedikaryotisation of dikaryons derived from five different monokaryons. When a single mechanism controls both processes, the nucleus better at fertilizing a monokaryon in a Buller pairing should also be recovered upon dedikaryotisation with a higher frequency. The results showed a hierarchical structure for recovery among nuclei in dedikaryotisation, but this hierarchy did not correspond to the fertilization success during di–mon mating. These findings thus show that the mechanism causing asymmetric regeneration of nuclei, is most likely not the same as the mechanism responsible for increased chance of fertilization in di–mon matings. We discuss the complexity of the interactions that occur during di–mon matings with regards to the mating type loci.     

Molecular cloning of RNAs differentially expressed in monokaryons and dikaryons ofSchizophyllum commune in relation to fruiting

A complementary DNA library cloned in pBR327 was prepared from RNA of a fruiting dikaryon of the basidiomyceteSchizophyllum commune. An improved colony-lysis method led to the isolation of complementary DNA clones hybridizing to nine different RNAs abundantly present in the fruiting dikaryon but present in very low or undetectable levels in the monokaryons. One of these RNAs, measuring 580 nucleotides, was exceptional in contributing more than 3% to the mRNA mass of the fruiting dikaryon. A role of the detected dikaryon-specific RNAs in fruiting was suggested by (i) their absence or low concentration in vegetatively growing monokaryons or the dikaryon, (ii) their steep increase in concentration during fruit-body formation in the dikaryon, (iii) a higher concentration of most of these RNAs in the fruit-body initials than in the supporting mycelium, and (iv) their prevalence in full-grown fruit bodies. The “fruiting-specific” RNAs were also present in a suspension-grown dikaryon, in the absence of fruiting, but in concentrations less than 10% of those in surface-grown fruiting dikaryon. The level of expression of these RNAs in the monokaryons was even lower or undetectable. Since the dikaryon and the monokaryons are otherwise coisogenic, it appears that the presence of two different incompatibility genes in the dikaryon allows for a low level of expression of the “fruiting genes” even in the absence of fruiting, whereas the genes are fully expressed under conditions permitting fruit-body formation.     

ABO blood groups and fertility—with special reference to intrauterine selection due to materno-fetal incompatibility

The purpose of the present investigation was to study whether there is differential fertility between different mating types of ABO blood group system. Selective force which is operating through maternal-fetal incompatibility has been observed in the differential fertility between compatible and incompatible mating groups in the present sample of 183 families of Visakhapatnam town of Andhra Pradesh, India. The differences in the mean numbers of pregnancies as well as living children between the two major mating groups, compatible and incompatible are significant. The fertility rates of O fathers and O mothers were significantly higher than those in matings in which neither parents belongs to O. The selection is operating to reduce the gene ratio of A and to increase the gene ratios of O and B in this sample.     

EVOLUTION OF VEGETATIVE INCOMPATIBILITY IN FILAMENTOUS ASCOMYCETES. I. DETERMINISTIC MODELS

In ascomycetes vegetative incompatibility can prevent the somatic exchange of genetic material between conspecifics. It must occur frequently in natural populations, since in all species studied many vegetative compatibility groups (VCGs) are found. Using a population-genetic approach, this paper explores two possible selective explanations for the evolution of vegetative incompatibility in asexual fungi: selection by a nuclear parasitic gene, and selection by a harmful cytoplasmic element. In a deterministic model, assuming a random spatial distribution of VCGs in an infinitely sized population, it is found that neither of these forms of frequency-dependent selection can explain the large number of VCGs found in nature. The selective pressure for more VCGs disappears once a limited number of VCGs exist, because the frequency of compatible interactions decreases when the number of VCGs increases. In comparing the two selective explanations, selection by a cytoplasmic element seems a more plausible explanation than selection by a nuclear gene.     

EVOLUTION OF INCOMPATIBILITY-INDUCING MICROBES IN SUBDIVIDED HOST POPULATIONS

Many insects, other arthropods, and nematodes harbor maternally inherited bacteria inducing "cytoplasmic incompatibility" (CI), reduced egg hatch when infected males mate with uninfected females. Although CI drives the spread of these microbes, selection on alternative, mutually compatible strains in panmictic host populations does not act directly on CI intensity but favors higher "effective fecundity," the number of infected progeny an infected female produces. We analyze the consequences of host population subdivision using deterministic and stochastic models. In subdivided populations, effective fecundity remains the primary target of selection. For strains of equal effective fecundity, if population density is regulated locally (i.e., "soft selection"), variation among patches in infection frequencies may induce change in the relative frequencies of the strains. However, whether this change favors stronger incompatibility depends on initial frequencies. Demographic fluctuations maintain frequency variation that tends to favor stronger incompatibility. However, this effect is weak; even with small patches, minute increases in effective fecundity can offset substantial decreases in CI intensity. These results are insensitive to many details of host life cycle and migration and to systematic outbreeding or inbreeding within patches. Selection acting through transfer between host species may be required to explain the prevalence of CI.     

BIPOLAR INCOMPATIBILITY IN THE HYMENOMYCETE POLYPORUS PALUSTRIS

The basic features of bipolar incompatibility in the hymenomycete, Polyporus palustris (Berk. and Curt.), have been investigated by a comparison of compatible and incompatible interactions leading to the formation of dikaryons and common-factor heterokaryons, respectively. Nuclear migration is severely restricted and nuclear pairing and conjugate nuclear division fail to occur in the interaction of incompatible homokaryons. It is suggested that the single incompatibility factor of P. palustris combines the functions of the two incompatibility factors of tetrapolar species.     

Paternity tests support a diallelic self‐incompatibility system in a wild olive (Olea europaea subsp. laperrinei,Oleaceae)

Self‐incompatibility (SI) is the main mechanism that favors outcrossing in plants. By limiting compatible matings, SI interferes in fruit production and breeding of new cultivars. In the Oleeae tribe (Oleaceae), an unusual diallelic SI system (DSI) has been proposed for three distantly related species including the olive (Olea europaea), but empirical evidence has remained controversial for this latter. The olive domestication is a complex process with multiple origins. As a consequence, the mixing of S‐alleles from two distinct taxa, the possible artificial selection of self‐compatible mutants and the large phenological variation of blooming may constitute obstacles for deciphering SI in olive. Here, we investigate cross‐genotype compatibilities in the Saharan wild olive (O. e. subsp. laperrinei). As this taxon was geographically isolated for thousands of years, SI should not be affected by human selection. A population of 37 mature individuals maintained in a collection was investigated. Several embryos per mother were genotyped with microsatellites in order to identify compatible fathers that contributed to fertilization. While the pollination was limited by distance inside the collection, our results strongly support the DSI hypothesis, and all individuals were assigned to two incompatibility groups (G1 and G2). No self‐fertilization was observed in our conditions. In contrast, crosses between full or half siblings were frequent (ca. 45%), which is likely due to a nonrandom assortment of related trees in the collection. Finally, implications of our results for orchard management and the conservation of olive genetic resources are discussed.     

DYNAMICS OF HYBRID INCOMPATIBILITY IN GENE NETWORKS IN A CONSTANT ENVIRONMENT

After an ancestral population splits into two allopatric populations, different mutations may fix in each. When pairs of mutations are brought together in a hybrid offspring, epistasis may cause reduced fitness. Such pairs are known as Bateson–Dobzhansky–Muller (BDM) incompatibilities. A well-known model of BDM incompatibility due to Orr suggests that the fitness load on hybrids should initially accelerate, and continue to increase as the number of potentially incompatible substitutions increases (the "snowball effect"). In the gene networks model, which violates a key assumption of Orr's model (independence of fixation probabilities), the snowball effect often does not occur. Instead, we describe three possible dynamics in a constant environment: (1) Stabilizing selection can constrain two allopatric populations to remain near-perfectly compatible. (2) Despite constancy of environment, punctuated evolution may obtain; populations may experience rare adaptations asynchronously, permitting incompatibility. (3) Despite stabilizing selection, developmental system drift may permit genetic change, allowing two populations to drift in and out of compatibility. We reinterpret Orr's model in terms of genetic distance. We extend Orr's model to the finite loci case, which can limit incompatibility. Finally, we suggest that neutral evolution of gene regulation in nature, to the point of speciation, is a distinct possibility.     

Identification and mode of action of self-compatibility loci in Lolium perenne L

The two-locus gametophytic incompatibility system in perennial ryegrass (Lolium perenne L.) is not always fully effective: obligate selfing of plants sieves self-compatible pollen mutants, and self-fertility becomes fixed in subsequent generations. Self-compatibility (SC) was investigated in an F2 family. In vitro self-pollinations were analysed and recorded and plants were classified as being either partially or fully compatible. Distorted segregation ratios of markers on linkage group (LG) 5 were found, which indicate the possible presence of a gametophytic SC locus. Interval linkage analysis of pollen compatibility after selfing confirmed that this distortion was due to a locus (T) analogous to the S5 locus of rye. However, even though markers in this region were, on average, less than 1 cM apart, the minimum number of plants possessing the unfavoured allele was never less than 6% for any marker locus. We proved that this was because of the presence of another SC locus, exhibiting gametophytic selection, segregating in this population and identified by interval mapping analysis of compatibility classes of in vitro self-pollinations. This locus was located on LG1, and probably corresponds to the S locus. We show that the T locus, a relic of a multilocus system, functions through interaction with the S locus: F2 segregation of incompatibility phenotypes and linked markers demonstrated that the S/t pollen genotype combination, expected to be compatible on selfing, was sometimes incompatible. Further evidence is presented to show that this interaction must be dependent on yet another locus located on LG2. A prime candidate would be the Z incompatibility locus.     

Cytological evidence for somatic diploidization in dikaryotic cells ofArmillariella mellea

Dikaryotic hyphae isolated from basidiocarps ofArmillariella mellea are unstable in aseptic culture and change into monokaryotic hypae. During monokaryotization the nuclei of a dikaryon fuse and fusion nucleus immediately divides resulting in two uninucleate cells, from which the monokaryotic mycelium originates. Similar fusion of two nuclei takes place in matings of compatible singlespore isolates. It is concluded that the resulting monokaryotic mycelium is diploid.     

Morphological mutation of Lentinula edodes mycelium, particularly detectable in the dikaryotic state

A morphological mutation particularly detectable in the dikaryotic state was found in Lentinula edodes. The mutant dikaryon was readily distinguishable from the normal dikaryon by the irregularly branched short hyphae, very slow hyphal growth, and sparse aerial hyphae. Genetic analysis revealed that expression of this mutation was controlled by a single recessive gene, mor-13. Linkage analysis showed that the mor-13 was not linked to either the incompatibility factors (A and B) or the five kinds of mor genes that were segregated independently of each other in a previous study. Contribution no. 380 from the Tottori Mycological Institute     

Parasexual analysis of common-A crosses in the basidiomycete Agrocybe aegerita

Summary Crosses were performed between homokaryons of Agrocybe aegerita having the same allele at the A incompatibility gene but different B alleles. Heterokaryotic mycelia originating from crosses between two complementary auxotrophs were characterized by their instability on complete medium and extensive anastomosis between hyphae. Diploid mycelia were selected by plating oidia recovered from these heterokaryons onto minimal medium. These mycelia were characterized by the production of larger oidia than those of homokaryons, the release of a brown pigment when growing on complete medium and extensive hyphal anastomoses. Diploids retained the two B incompatibility functions of their homokaryotic parents and gave rise to a diploid/haploid dikaryon when crossed with a compatible homokaryon. Nearly 1% of the oidia recovered from heterokaryons were diploid. These nuclear fusion frequencies as well as the production of brown pigments enabled the identification of diploid strains on complete medium. In this way, crosses between wild prototrophic strains were successfully performed. Somatic recombination was induced following the treatment of diploid mycelia with haploidizing compounds. Selection based on the inability of mycelia to produce the brown pigments on complete medium led to selection of strains homoallelic at the B locus.     

The S-locus and unilateral incompatibility

Plants have many ways to regulate the type of pollen that arrives on the stigma surface. Once there, further control mechanisms regulate compatibility. The latter controls are largely based on biochemical interactions that support compatible pollination and prevent incompatible matings. S-RNase-based self-incompatibility (SI) systems are the most phylogenetically widespread mechanisms for controlling pollination. Studies of Nicotiana establish a firm link between SI and unilateral interspecific incompatibility. Although implicated in both inter- and intraspecific compatibility, S-RNase operates through at least three distinct genetic mechanisms that differ in their dependence on non-S-RNase factors. Identification and characterization of these non-S-RNase factors is currently an area of active research. Searching for genetic and biochemical interactions with S-RNase can identify candidate non-S-RNase factors. HT-protein is one factor that is required for S-allele-specific pollen rejection in the Solanaceae. Major style arabinogalactan proteins such as TTS interact biochemically with S-RNase. These glycoproteins are known to interact with compatible pollen tubes and have long been suggested as possible recognition molecules. Their binding to S-RNase implies a link between stylar systems for compatibility and incompatibility. Thus, genetic and biochemical studies suggest a highly networked picture of pollen-pistil interactions.     

Sperm numbers vary between inter- and intra-population matings of the grasshopper Chorthippus parallelus

Comparing the reproductive output of intra- and inter-population matings is the most common way to assess whether post-mating reproductive isolation is caused by genetic incompatibilities. Such genetic incompatibility can however, only assume that the quantity of the post-mating signals involved does not differ between intra- and inter-population matings. This assumption may not be true because sexual selection predicts reduced mating effort towards low-quality mates and in many circumstances, allopatric partners are low-quality mates. Post-mating efforts may, therefore, be reduced in inter- compared to intra-population matings. Here, I test this crucial assumption by studying variation in one post-mating trait, sperm number, in crosses of two parapatric grasshopper populations. In both populations, males transferred fewer sperm to allopatric than sympatric females. If such plasticity with respect to population is common in other post-mating traits, differences between inter- and intra-population crosses may be more frequently caused by differences in sperm number rather than gamete incompatibility. Additionally, I found that sperm numbers declined less rapidly in the female storage organ of allopatric than sympatric females but its rate differed markedly between populations. This is discussed with respect to female adaptations to male traits.