Intraspecific and within-isolate sequence variation in the ITS rRNA gene region of Pythium mercuriale sp. nov. (Pythiaceae).

Sixteen Pythium isolates from diverse hosts and locations, which showed similarities in their morphology and sequences of the internal transcribed spacer (ITS) region of their rRNA gene, were investigated. As opposed to the generally accepted view, within single isolates ITS sequence variations were consistently found mostly as part of a tract of identical bases (A-T) within ITS1, and of GT or GTTT repeats within the ITS2 sequence. Thirty-one different ITS sequences obtained from 39 cloned ITS products from the 16 isolates showed high sequence and length polymorphisms within and between isolates. However, in a phylogenetic analysis, they formed a cluster distinct from those of other Pythium species. Additional sequencing of two nuclear genes (elongation factor 1 alpha and beta-tubulin) and one mitochondrial gene (nadh1) revealed high levels of heterozygosity as well as polymorphism within and between isolates, with some isolates possessing two or more alleles for each of the nuclear genes. In contrast to the observed variation in the ITS and other gene areas, all isolates were phenotypically similar. Pythium mercuriale sp. nov. (Pythiaceae) is characterized by forming thin-walled chlamydospores, subglobose to obovoid, papillate sporangia proliferating internally and smooth-walled oogonia surrounded by multiple antheridia. Maximum likelihood phylogenetic analyses based on both ITS and beta-tubulin sequence data place P. mercuriale in a clade between Pythium and Phytophthora.

A new species of Pythium with ornamented oogonia: morphology, taxonomy, internal transcribed spacer region of its ribosomal RNA, and its comparison with related species.

Pythium spiculum sp. nov. was isolated from soil samples taken in a vineyard in the Burgundian region of France and from different locations in Spain and Portugal. The oomycete has spiny oogonia and does not sporulate readily. It resembles Pythium mamillatum Meurs, but has its own distinguishing characteristics. It also exhibits sickle-shaped as well as spherical appressoria which at times are associated with sex organs like those found in Pythium abappressorium Paulitz and Pythium contiguanum Paul. Sequencing of the internal transcribed spacer region of its nuclear ribosomal DNA and a close look at its morphological characters have now enabled us to describe it as a new species. The internal transcribed spacer region of its rRNA gene sequence is comprised of 945 bases. This oomycete is closely related to the members that form ornamented or spiny oogonia like Pythiummamillatum, Pythium spinosum and Pythium irregulare but also with those producing smooth-walled oogonia like Pythium paroecandrum, Pythium sylvaticum and Pythium cylindrosporum. Taxonomic description of this new species, its comparison with related oomycetes, the sequence of the internal transcribed spacer region of its rRNA gene and the phylogenetic tree, are given here.

Pythium sterilum sp. nov. isolated from Poland, Spain and France: its morphology and molecular phylogenetic position.

In a survey of Phytophthora species associated with forest decline in Spain, Poland and France, we found three Pythium isolates, which have been characterized with internal transcribed spacer rRNA gene sequences and with classical morphological descriptors for Pythium spp. These isolates showed unique internal transcribed spacer sequences, different enough from those of any described species to justify new species status. These three distinct isolates failed to produce any sex organs with an entirely asexual reproduction and were found to represent a new species for which the name Pythium sterilum is proposed. This paper describes and illustrates the morphology of P. sterilum and presents its taxonomic position and relationships with other, related Pythium species belonging to clade K.

Phytophthora inundata sp. nov., a part heterothallic pathogen of trees and shrubs in wet or flooded soils.

A Phytophthora pathogen of trees and shrubs previously designated Phytophthora sp. O-group is formally named as P. inundata sp. nov. P. inundata falls within the P. gonapodyides-P. megasperma major ITS Clade 6, its present nearest known relative being P. humicola. It has non-papillate sporangia, fairly large oogonia (average ca 40 microns) with thick walled oospores, amphigynous antheridia, a distinctive colony type, a high optimum temperature for growth of 28-30 degrees C, fast growth at the optimum, and a high upper temperature limit for growth of ca 35-37 degrees. A study of the breeding system of eight P. inundata isolates showed them to be classically heterothallic with A1 and A2 compatibility types. However some P. inundata A1 x A2 combinations failed to mate even though the same isolates mated successfully with P. drechsleri testers. Others were 'silent' A1s or A2s, unable to produce their own gametangia but able to induce gametangial formation in the opposite sexual compatibility type of another species. This indicates a partial breakdown of the sexual mechanism in the species. Two isolates (one A1 and one A2) were unpredictably and chimaerically self-fertile, suggesting A1 + A2 chromosomal heteroploidy. The association of P. inundata with ponds and rivers and with root and collar roots of trees and shrubs after flooding is discussed.

Identity and Pathogenicity of Two Phytophthora Taxa Associated with a New Root Disease of Olive Trees.

Fifty-two Phytophthora isolates from necrotic roots of olives were characterized. Colony morphologies on carrot-agar medium led us to separate them into two groups: A (36 isolates) and B (16 isolates). The optimum growth temperature for Group A was about 21°C, with slow growth at 30°C. In contrast, Group B isolates had an optimum temperature for growth of 26°C, and grew rapidly at 30°C. Growth rates, sporangial and oogonial characteristics of the Group A isolates conformed to P. megasperma "BHR-type" sensu stricto. This designation was supported by a sequence analysis of their ITS rDNA regions. Colony patterns, sporangial characteristics and temperature-growth relationships of the Group B isolates conformed closely to those of the 'O-group' taxon of Phytophthora. They also conformed to this unusual taxon in their ITS sequence. In addition, Group B isolates were either entirely self-sterile, self-sterile A1s or weakly self-fertile. Pathogenicity tests showed that both taxa were highly aggressive on roots of olive trees. The association of flooding with Phytophthora infection indicates that the previously reported high sensitivity of olive to root asphyxiation may be more properly regarded as root-rot caused by Phytophthora spp.