Jane M. Bowles

Metschnikowia lochheadii and Metschnikowia drosophilae, two new yeast species isolated from insects associated with flowers.

Two new haplontic heterothallic species of Metschnikowia were isolated from floricolous insects and flowers. Metschnikowia lochheadii was recovered from insects found in various flowers on the Hawaiian Islands of Kauai and Maui, and from Conotelus sp. (Coleoptera: Nitidulidae) in northwestern Guanacaste Province, Costa Rica. The morphology, physiology, and sexual cycle are typical of the large-spored Metschnikowia species, and the partial ribosomal DNA large subunit (D1D2) sequences suggest that the new species is most closely related to Candida ipomoeae. Metschnikowia lochheadii is nearly indistinguishable from its ascogenous relatives and conjugates freely with Metschnikowia continentalis, forming sterile asci. It also exhibits asymmetric mating with Metschnikowia hawaiiensis. Metschnikowia drosophilae was found in morning glory (Ipomoea sp.) flowers and associated Drosophila bromeliae on Grand Cayman Island. Its nutritional profile is atypical of the genus, being the only species that does not utilize sucrose or maltose as carbon sources, and one of the few that does not utilize melezitose. D1D2 sequences show that Metschnikowia drosophilae is a sister species to Candida torresii, to which it bears considerable similarity in nutritional profile. The type cultures are: Metschnikowia lochheadii, strains UWO(PS)00-133.2 = CBS 8807 (h+, holotype) UWO(PS)99-661.1 = CBS 8808 (h-, isotype); and Metschnikowia drosophilae, strains UWO(PS)83-1135.3 = CBS 8809 (h+, holotype) and UWO(PS)83-1143.1 = CBS 8810 (h-, isotype).

A study of the effects of trampling on the vegetation of Lake Huron sand dunes at Pinery Provincial Park

Abstract Plots in established heath-grassland vegetation and pure stands of Calamovilfa longifolia (Hook) Scribn. on Lake Huron sand dunes were subjected to tramppling treatments at various intensities during 1977 and 1978 and were monitored for one year following treatment. Both heath-grassland and Calamovilfa plots given 50 walking passages showed complete recovery within one year after treatment. Heavier treatments (200 walking passages) on Calamovilfa caused a significant reduction in cover one year later. Heavily trampled heath-grassland plots, although visually similar to their controls, had a continued loss of lichen cover over one year. Several species including Arctostaphylos uva-ursi (L.) Spreng., Andropogon scoparius Michx., some Carex species, Liatris cylindracea Michx. and Rosa blanda Ait. initiated new growth in the fall following heavy trampling treatment, but only shoots of Arctostaphylos survived the winter.

Kodamaea nitidulidarum, Candida restingae and Kodamaea anthophila, three new related yeast species from ephemeral flowers

Three new yeast species were discovered during studies of yeasts associated with ephemeral flowers in Brazil, Australia and Hawaii. Their physiological and morphological similarity to Kodamaea (Pichia) ohmeri suggested a possible relationship to that species, which was confirmed by rDNA sequencing. Kodamaea nitidulidarum and Candida restingae were found in cactus flowers and associated nitidulid beetles in sand dune ecosystems (restinga) of South-eastern Brazil. Over 350 strains of Kodamaea anthophila were isolated from Hibiscus and morning glory flowers (Ipomoea spp.) in Australia, and from associated nitidulid beetles and Drosophila hibisci. A single isolate came from a beach morning glory in Hawaii. Expansion of the genus Kodamaea to three species modified the existing definition of the genus only slightly. The type and isotype strains are as follows: K. nitidulidarum strains UFMG96-272T (h+; CBS 8491T) and UFMG96-394I (h-; CBS 8492I); Candida restingae UFMG96-276T (CBS 8493T); K. anthophila strains UWO(PS)95-602.1T (h+; CBS 8494T), UWO(PS)91-893.2I (h-; CBS 8495I) and UWO(PS)95-725.1I (h-; CBS 8496I).

Wickerhamiella australiensis, Wickerhamiella cacticola, Wickerhamiella occidentalis, Candida drosophilae and Candida lipophila, five new related yeast species from flowers and associated insects

Five new yeast species, Wickerhamiella australiensis, Wickerhamiella cacticola, Wickerhamiella occidentalis, Candida drosophilae and Candida lipophila, are described to accommodate isolates recovered from flowers and floricolous insects of Australian Hibiscus trees, cosmopolitan morning glories (Ipomoea spp.) and Brazilian cereoid cacti. The new Wickerhamiella species are heterothallic, occur in the haploid condition and are clearly separated reproductively from one another. Although they exhibit little physiological variation, they are easily delineated from Wickerhamiella domercqiae, the only species known previously, by their resistance to cycloheximide and the production of strong extracellular lipases. C. drosophilae and C. lipophila share the latter property, but unlike the Wickerhamiella species, they fail to utilize nitrate as sole nitrogen source. PFGE indicates that these yeasts have an unusually low number of chromosomes. The large-subunit rDNA (D1/D2) sequences demonstrate a close relationship between the five species and Candida vanderwaltii and Candida azyma. Their relationship with W. domercqiae is more distant, but all share, with some other Candida species, a single monophyletic clade. The type and isotype strains are as follows: W. australiensis strains UWO(PS)95-604.3T (h+; CBS 8456T) and UWO(PS)95-631.3I (h-; CBS 8457I); W. cacticola strains UFMG96-267T (h+; CBS 8454T) and UFMG96-381I (h-; CBS 8455I); W. occidentalis strains UWO(PS)91-698.4T (h+; CBS 8452T) and UFMG96-212I (h-; CBS 8453I); C. drosophilae UWO(PS)91-716.3T (CBS 8459T); and C. lipophila UWO(PS)91-681.3T (CBS 8458T).

Ribosomal DNA, species structure, and biogeography of the cactophilic yeast Clavispora opuntiae

The ribosomal DNA of the cactophilic yeast species Clavispora opuntiae was studied in order to clarify the global distribution of the yeast. Over 500 strains, including isolates from several new localities worldwide, were characterized by rDNA restriction mapping. An unusual restriction pattern previously encountered only in one strain, from Conception Island in the Bahamas, was found in several Brazilian isolates. Sequences of the D1/D2 and D7/D8 divergent domains of the large subunit (LSU) and of the intergenic spacers (IGS) confirmed that these strains represent a genetically distinct variety of Clavispora opuntiae. This divergence had previously been hypothesized on the basis of reduced genetic recombination in inter-varietal crosses and the presence of a polymorphic ApaI restriction site located in the LSU. The exact position of the ApaI site in the D8 divergent domain and the nature of the variation that it reveals were determined. The complete sequences of 12 intergenic spacers clarified the significance of the species-wide variation uncovered by restriction mapping. Most of the polymorphic sites occur in the IGS1 and IGS2 regions, on either side of the 5S gene, and the variation is largely due to differences in the numbers and the sequences of internal repeats. Two other polymorphic sites are located in the external transcribed spacer (ETS) region. The reliability of various sites as indicators of overall spacer sequence divergence differed from one case to another. Variety-specific probes were devised and used to screen 120 strains for the presence of recombinant rDNA spacers. Three strains gave ambiguous results, but these did not constitute evidence that inter-varietal recombination has taken place in nature. The hypothesis that the global movement of Clavispora opuntiae has been influenced by the worldwide biological control of prickly pear with Cactoblastis cactorum, a moth of Argentinian origin, has received additional support from the demonstration that Argentinian strains have rDNAs similar to those found where the moth has been introduced. A dramatic founder effect was identified in a yeast population collected in cacti (Maui, Hawaii) in a site where the moth had been recently introduced.

On the biogeography of yeasts in the Wickerhamiella clade and description of Wickerhamiella lipophila sp. nov., the teleomorph of Candida lipophila.

We describe the new yeast species Wickerhamiella lipophila, the teleomorph of Candida lipophila, a haploid heterothallic yeast previously isolated from insects associated with morning glories in Hawaii. Both mating types were recovered in the eastern region of Maui, and a single strain was found in the Waimea region of Kauai. We reexamined the mating compatibility of the several strains of Candida lipophila previously collected on the island of Hawaii and found them to be fertile mating types that had been overlooked because of the unpredictability of mating and ascus formation. The type culture of Candida lipophila [UWO(PS)91-681.3 = CBS 8458, h+] is transferred to the genus Wickerhamiella, and strain UWO(PS)00-340.1 (CBS 8812, h-) is designated as isotype. Also found on Maui and Kauai were strains of Candida drosophilae that produced a strong extracellular protease. An update on the global distribution of members of the Wickerhamiella clade is given.

Saccharomycopsis fodiens sp. nov., a rare predacious yeast from three distant localities.

Three strains representing a novel yeast species were recovered as part of independent collections from flower-associated nitidulid beetles in Australia, Costa Rica and the Galapagos Islands, Ecuador. Analysis of the D1/D2 domains of the large subunit rRNA gene indicated that the species belongs to the genus Saccharomycopsis, although the formation of ascospores was not observed. The yeast is capable of necrotrophic parasitism by means of infection pegs when mixed with other yeasts or filamentous fungi. Of particular interest is the fact that despite the large distances separating the isolation sites of the three strains, other strains of the species have not been recovered in other samples of flower-associated nitidulids even though these habitats have been sampled extensively. It is suggested that the dispersal of the yeast may be linked to human historical factors. The name Saccharomycopsis fodiens sp. nov. is proposed for the yeast. The type strain is UWOPS 95-697.4(T) (=CBS 8332(T)=NRRL Y-48786(T)).

Metschnikowia shivogae sp. nov., a yeast species associated with insects of morning glory flowers in East Africa

The novel species Metschnikowia shivogae is described to accommodate three isolates recovered from insects of morning glory flowers at two localities in East Africa. The isolates differ slightly in rDNA ITS and D1/D2 large-subunit sequences and one isolate featured a two-base heterogeneity that might be the result of recombination between two variant rDNAs. M. shivogae is a sister species to Metschnikowia aberdeeniae and shares the same habitat. The reproductive boundaries of M. aberdeeniae, which were not clear in the past, have now been elucidated further. The type strain of Metschnikowia shivogae sp. nov. is strain SUB 04-310.1(T) (h(+); =CBS 10292(T) =NRRL Y-27924(T)) and the allotype is strain UWOPS 07-203.2 (h(-); =CBS 10770 =NRRL Y-48447).