Worker Size, Geographical Distribution, and Introgressive Hybridization of Invasive Solenopsis invicta and Solenopsis richteri (Hymenoptera: Formicidae) in Tennessee.

Worker size and geographical distribution of red imported fire ants (Solenopsis invicta Buren), black imported fire ants (Solenopsis richteri Forel), and their hybrid (S. invicta × S. richteri) (Hymenoptera: Formicidae) were evaluated from colonies sampled across Tennessee. The fire ant species and hybrid status were determined using cuticular hydrocarbon and venom alkaloid indices obtained from gas chromatography and mass spectrometry. Hybrids were the most common fire ant throughout Tennessee. With the exception of a few isolated S. invicta samples, only hybrids were found in east Tennessee, and hybrids predominated in middle Tennessee. In west Tennessee, mixed populations of S. richteri and hybrids were found. Hybrids were more common in west Tennessee than a survey performed a decade earlier. No statistical differences were detected in the average inter-colonial worker size of S. richteri and hybrids. Likewise, average worker size was not related to geographic location in Tennessee. The similarity in average worker size among hybrid colonies with a wide range of cuticular hydrocarbon and venom alkaloid values suggests introgression was not impacting ant size in colonies sampled throughout Tennessee.

Candida orba sp. nov., a new cactus-specific yeast species from Queensland, Australia.

A new species of yeast from decaying cladodes of Opuntia cactus, Candida orba, is described. This species is a member of a four-species clade of cactophilic yeasts. The new species has only been found in one region of Queensland, Australia, where it was presumably introduced during attempts to eradicate prickly pear cactus. DNA-DNA relatedness, phylogenetic analysis, physiological differences, killer-sensitivity profiles and mating reactions establish the distinctness of the taxon as a new species. C. orba is most closely related to Phaffomyces thermotolerans, a species found associated with columnar cacti in the North American Sonoran Desert. The type strain of C. orba, isolated from rotting cladodes of Opuntia stricta in the State of Queensland, Australia, is strain UCD-FST 84-833.1T (= CBS 8782T = NRRL Y-27336T = ATCC MYA-341). Only the h- mating type of the species has been recovered. The lack of the opposite mating type could be the result of a bottleneck during its introduction to Australia. The original geographic/host distribution of this species in the Americas is unknown.

The use of anonymous DNA markers in assessing worldwide relatedness in the yeast species Pichia kluyveri Bedford and Kudrjavzev.

Pichia kluyveri, a sexual ascomycetous yeast from cactus necroses and acidic fruit, is divided into three varieties. We used physiological, RAPD, and AFLP data to compare 46 P. kluyveri strains collected worldwide to investigate relationships among varieties. Physiology did not place all strains into described varieties. Although the combined AFLP and RAPD data produced a single most parsimonious tree, separate analysis of AFLP and RAPD data resulted in significantly different trees (by the partition homogeneity test). We then compared the distribution of strains per band to an expected distribution. This suggested we could separate both the AFLP and RAPD datasets into bands from rapidly and slowly changing DNA regions. When only bands from slowly changing regions (from each dataset) were included in the analysis, both the RAPD and AFLP datasets supported a single tree. This second tree did not differ significantly from the cladogram based on all of the DNA data, which we accepted as the best estimate of the phylogeny of these yeast strains. Based on this phylogeny, we were able to demonstrate the strong influence of geography on the population structure of this yeast, confirm the monophyly of one variety, question the utility of maintaining another variety, and demonstrate that the physiological differences used to separate the varieties did not do so in all cases.

Analysis of population structure of cactophilic yeast from the genus Pichia: P. cactophila and P. norvegensis.

DNAs from 40 strains of Pichia cactophila and Pichia norvegensis, yeasts characteristic of cactus necroses, were compared using randomly amplified polymorphic DNA (RAPD) banding patterns and killer/sensitive phenotypes. Both species belong to the same species complex within the genus. The levels of between-strain RAPD variation were high in both species (higher in the automictic P. cactophila than in the heterothallic P. norvegensis), although there is little variation in physiological abilities within either species. Although each species was a separate lineage, RAPD analysis confirms that the species are related. Within each species, RAPD variation was related to the geographic origin of the strains. Pichia cactophila strains from southern Florida were more related to those from Antigua than to those from northern Florida. These results correlated well with the differences among killer/sensitive phenotypes of strains. Principal component analysis indicated that the phenotypes of each species differ. Here too, strains from southern Florida were more similar to those from Antigua than to those from northern Florida. Previous work had identified differences in the cactophilic yeast communities from southern and northern Florida, and these results indicate that the differences are detectable at the population levels as well.

Candida caseinolytica sp. nov., a new species of yeast occurring in necrotic tissue of Opuntia and Stenocereus species in the southwestern United States and Baja California, Mexico.

We describe Candida caseinolytica, a new yeast species which occurs in rotting tissues of opuntias and other cacti in the North American Sonoran Desert and a few other localities. This small-celled, slowly growing yeast does not ferment any sugar and assimilates a limited number of carbon compounds, including 2- and 5-ketogluconic acids. It exhibits strong extracellular proteolytic activity on casein at pH 6.5, but gelatin is not hydrolyzed or is only weakly hydrolyzed by a few strains. The type strain of C. caseinolytica is strain UCD-FST 83-438.3 (= ATCC 90546 = CBS 7781).

Geographic distribution and genetics of killer phenotypes for the yeast Pichia kluyveri across the United States.

Representative strains (n = 61) of the yeast Pichia kluyveri from across the United States were studied for their ability to kill 71 other strains (representing 25 species) of yeast. This survey showed killing activity in 69% of the P. kluyveri strains tested. More extensive analysis of killer activity of 197 P. kluyveri strains against strains of five tester species showed comparable activity (67% of strains tested). This activity was shown to be equally variable within localities, within regions, and across the continent. The genetic basis of the variability was ascertained by tetrad analysis and is most likely due to alleles segregating at three epistatic loci. Evidence for the idea that killer toxins have a role in excluding other yeasts from particular habitats is discussed.

The ecological role of killer yeasts in natural communities of yeasts.

The killer phenomenon of yeasts was investigated in naturally occurring yeast communities. Yeast species from communities associated with the decaying stems and fruits of cactus and the slime fluxes of trees were studied for production of killer toxins and sensitivity to killer toxins produced by other yeasts. Yeasts found in decaying fruits showed the highest incidence of killing activity (30/112), while yeasts isolated from cactus necroses and tree fluxes showed lower activity (70/699 and 11/140, respectively). Cross-reaction studies indicated that few killer-sensitive interactions occur within the same habitat at a particular time and locality, but that killer-sensitive reactions occur more frequently among yeasts from different localities and habitats. The conditions that should be optimal for killer activity were found in fruits and young rots of Opuntia cladodes where the pH is low. The fruit habitat appears to favor the establishment of killer species. Killer toxin may affect the natural distribution of the killer yeast Pichia kluyveri and the sensitive yeast Cryptococcus cereanus. Their distributions indicate that the toxin produced by P. kluyveri limits the occurrence of Cr. cereanus in fruit and Opuntia pads. In general most communities have only one killer species. Sensitive strains are more widespread than killer strains and few species appear to be immune to all toxins. Genetic study of the killer yeast P. kluyveri indicates that the mode of inheritance of killer toxin production is nuclear and not cytoplasmic as is found in Saccharomyces cerevisiae and Kluyveromyces lactis.