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.     

First records of Pythium aquatile and P. macrosporum isolated from soils in Japan

Pythium aquatile and P. macrosporum were isolated from the soil of a cultivated field in Gunma Prefecture and a forest in Nagano Prefecture for the first time in Japan. Their morphological characteristics are described, and their pathogenicity and taxonomy are discussed.     

Flavonoids in roots of white clover: interaction of arbuscular mycorrhizal fungi and a pathogenic fungus

The effects of two arbuscular mycorrhizal fungi (AMF) (Glomus mosseae and G. claroideum) and a pathogenic fungus (Pythium ultimum) on the production of eight flavonoids in roots of two white clover (Trifolium repens L.) cultivars were evaluated. Quantification of AM and pathogenic fungi in the roots showed that the AM symbiosis significantly reduced P. ultimum biomass and in some cases prevented infection. The flavonoid productions in clover roots varied depending on the presence of beneficial and/or pathogenic fungi, fungal isolate or plant cultivar. Only plants colonized with G. claroideum showed detectable concentrations of either coumestrol or kaempferol (cultivar-dependant). In addition, inoculation with G. claroideum resulted in significantly higher concentrations of coumestrol in cv. Sonja and medicarpin in cv. Milo. A low production of coumestrol and kaempferol in mycorrhizal plants may be G. mosseae-specific. Only the concentrations of formononetin and daidzein increased in clover roots in response to infection with P. ultimum. These flavonoids are supposedly stress metabolites, synthesized or produced from glycosides in response to pathogen infection. However, the presence of one or both AMF significantly lowered the formononetin and daidzein concentrations, and overruled the inductive effect of P. ultimum. Therefore the antagonistic action of AM against the pathogen must take place through another mechanism.     

Serological detection of red rot disease initiation stages of microbial pathogen, Pythium porphyrae (Oomycota) on Porphyra yezoensis

Pythium porphyrae (Oomycota), a pathogen causing red rot diseasein Porphyra spp., can at present only be detected when colonizationof the host thallus has already occurred and so it is often too late to takeappropriate disease control measures. The paper presents an account of an effective methdology for early detection of the disease. Since Py.porphyrae zoospores are the primary means of pathogen dispersal,polyclonal antibodies (Pabs) were raised against the surface components ofzoospores and encysted zoospores. Using these Pabs the disease initiationstages of the Pythium porphyrae were detected on the surface of Porphyra thalli by immunofluorescence assay. The specificity of theseantibodies and the efficacy of immunofluorescence assay in the detectionof red rot disease are discussed.     

莴苣叶疫病和茎腐病的病原

2021年3–6月,从甘肃省的生菜(叶用莴苣)叶片和青海省的莴笋(茎用莴苣)茎秆罹病样本上分离得到腐霉属卵菌。通过Koch’s法则明确了分出菌株的致病性。依据形态学和分子生物学特征,将3个菌株鉴定为嗜导管腐霉Pythium tracheiphilum。在核糖体DNA内转录间隔区(rDNA-ITS)、细胞色素氧化酶亚基1(cox1)和核糖体DNA28S大亚基(rDNA-LSU)基因联合系统发育树中,甘肃菌株(LPy-B)和青海菌株(LPy-C和LPy-D)被聚在P. tracheiphilum的不同亚群里,不同菌株的适宜生长温度和产孢特性存在差异。孢子囊顶生、间生或侧生,球形,17.13–53.73μm,或近球形至葫芦状,24.58–56.72×18.62–53.73μm;休止孢球形,6.70–9.68μm;藏卵器光滑,顶生或间生,球形,15.64–23.09μm;每个藏卵器有雄器1–2个,雄器与藏卵器同丝或异丝生;卵孢子满器或近满器,球形,直径13.41–20.11μm,卵孢子壁厚0.74–2.23μm。致病性测定结果表明,除莴苣外,嗜导管腐霉还可侵染菊科的华蒲公英和刺儿菜、十字花科的...     

Interaction of the mycoparasite Pythium oligandrum with other Pythium species

Interactions of Pythium oligandrum and four plant‐pathogenic Pythium spp. (P. ultimum, P. vexans, P. graminicola and P. aphanidermatum,) were studied in vitro by (i) video microscopy of hyphal interactions on water agar films, (ii) counting of host and mycoparasite propagules in different regions of opposing colonies on sunflower‐seed extract agar films and (Hi) ability of P. oligandrum to overgrow plates of potato‐dextrose agar previously colonized by Pythium spp. Pythium oligandrum typically coiled round the hyphae of Pythium hosts and penetrated the host hyphae after approximately 50 min from the hyphal coils, causing disruption of host hyphal tips up to 1.2 mm ahead of contact points. The relative growth rates of mycoparasite and host hyphae, timing of penetration and distance (sub‐apical) at which penetration led to host tip disruption were used to assess the potential of mycoparasitism by P. oligandrum to prevent the growth of Pythium hosts. P. aphanidermatum was unique among the ‘host’ Pythium spp. in being largely unaffected by P. oligandrum and in antagonizing the mycoparasite by coiling and penetrating the mycoparasite hyphae. Other host Pythium spp. apparently differed in susceptibility, the most susceptible being P. vexans and P. ultimum, whereas P. graminicola was more resistant. The results are discussed in relation to the role of P. oligandrum as a biocontrol agent, especially for limiting the ability of other Pythium spp. to increase their propagule populations in crop residues.     

Characterization of two morphological groups of isolates ofPythium ultimum var.ultimum in a vegetable field

Comparisons were made between two morphological groups ofPythium ultimum var.ultimum strains isolated in a vegetable field in Japan. The groups were distinguished as having smaller or larger sexual organs by the sizes of their antheridia and oogonia. Morphological study indicated that the two groups comprised a single taxon,P. ultimum var.ultimum, by the current taxonomical keys. The smaller group grew faster in the lower temperature range of 4–15°C, whereas the larger group grew faster in the higher temperature range of 25–37°C. Random amplified polymorphic DNA (RAPD) and isozyme analyses revealed genetic dissimilarity between the two groups. Cluster analysis of the isozyme banding patterns with four otherPythium spp. demonstrated that the genetic dissimilarity between the two groups was equivalent to species level. In the field survey, the smaller group was frequently detected in February, May and September but not in July, while the larger group was detected mainly in July and September. The two groups were not distinguishable by their pathogenicity to spinach seedlings.     

Chemical,physical and biological control of carrot seedling diseases

In naturally infested soil containingPythium ultimum, P. acanthicum andPhytophthora megasperma, onlyP. ultimum was associated with root rot and damped-off seedlings. Damping-off was promoted by low soil temperatures and by flooding. Seedling stands were markedly reduced when seed was pre-incubated in soil at 12°C but not at 25°C or 35°C. Dusting carrot seed with metalaxyl significantly increased seedling stands in the field at rates from 1.5–6 g kg⁻¹ seed and in both flooded and unflooded, naturally infested soil at 3.15 g kg⁻¹. In greenhouse experiments using artifically infested soil,P. ultimum andP. paroecandrum caused damping-off of carrot seedlings andRhizoctonia solani reduced root and shoot weights.R. solani caused damping-off in nutrient-enriched soil.P. acanthicum andP. megasperma were not pathogenic to seedlings, although both fungi colonized roots. Soil populations of allPythium spp., particularlyP. ultimum, increased during growth of seedlings and population growth ofP. megasperma was promoted by periodic flooding. Infestation of soil withP. acanthicum did not reduce damping-off of carrot seedlings byP. ultimum orP. paroecandrum, but significantly increased root and shoot weights and decreased root colonization byR. solani P. acanthicum has potential as a biocontrol agent againstR. solani.     

中国腐霉属的生态和分布

腐霉属PytMum真菌全世界已报道约100种。本文记载我国的腐霉45种,以及它们的生态及地理分布。腐霉喜栖息于富含有机质的潮湿土壤里,以菜园土中为最多,其次是大田土、水畔土和陵园土,干旱的沙质土、森林土和含盐碱的沼泽土一般均较少,最贫乏的是草丘土,如草原、牧场、山坡、荒丘及道旁等土壤。在我国的土壤中,以绚丽腐霉P．Pulchrum和瓜果腐霉P. aphanidermatum最为常见,其次是刺腐霉Spinosum、中国腐霉P．Sinense和卡地腐霉P. carolinianum,而顶生腐霉P．Acrogenum、色孢腐霉P．Coloratum、壁台腐霉P connatura等10个种却较罕见,仅出现过一次。腐霉在土壤中的垂直分布以5—10㈨深处为最多。腐霉在土壤中的活动有着明显的季节性变化：春季最多,秋季次之,夏季最少。在我国的30个省市自治区中,北京发现的腐霉最多,已报道22种;河北、广东、云南省次之,各有13种;台湾省已记录12个种;山西省、甘肃省和新疆维吾尔自治区均仅发现一个种;而青海省、宁夏回族自治区和西藏自治区迄今尚未见有腐霉报道。     

Analogous wheat root rhizosphere microbial successions in field and greenhouse trials in the presence of biocontrol agents Paenibacillus peoriae SP9 and Streptomyces fulvissimus FU14

Two Pythium-infested soils were used to compare the wheat root and rhizosphere soil microbial communities from plants grown in the field or in greenhouse trials and their stability in the presence of biocontrol agents. Bacteria showed the highest diversity at early stages of wheat growth in both field and greenhouse trials, while fungal diversity increased later on, at 12 weeks of the crop cycle. The microbial communities were stable in roots and rhizosphere samples across both soil types used in this study. Such stability was also observed irrespective of the cultivation system (field or greenhouse) or addition of biocontrol coatings to wheat seeds to control Pythium disease (in this study soil infected with Pythium sp. clade F was tested). In greenhouse plant roots, Archaeorhizomyces, Debaryomyces, Delftia, and unclassified Pseudeurotiaceae were significantly reduced when compared to plant roots obtained from the field trials. Some operational taxonomic units (OTUs) represented genetic determinants clearly transmitted vertically by seed endophytes (specific OTUs were found in plant roots) and the plant microbiota was enriched over time by OTUs from the rhizosphere soil. This study provided key information regarding the microbial communities associated with wheat roots and rhizosphere soils at different stages of plant growth and the role that Paenibacillus and Streptomyces strains play as biocontrol agents in supporting plant growth in infested soils.