Draft genome sequences of Chryseobacterium sp. MHB01, Rhodococcus qingshengii MHB02, and Agrobacterium tumefaciens MHB03 isolated from superabsorbent polymer granules inoculated with an arbuscular mycorrhizal fungus.

Chryseobacterium sp. MHB01, Rhodococcus qingshengii MHB02, and Agrobacterium tumefaciens MHB03 were isolated from superabsorbent polymer granules cultured with an arbuscular mycorrhizal fungus. Whole-genome sequencing of these three strains revealed genome sizes of 4.57 Mb, 7.13 Mb, and 5.49 Mb with G + C contents of 36.9%, 62.5%, and 58.2%, respectively.

Rhizophagus irregularis, the model fungus in arbuscular mycorrhiza research, forms dimorphic spores.

Rhizophagus irregularis is the model species for arbuscular mycorrhizal fungi (AMF) research and the most widely propagated species for commercial plant biostimulants. Using asymbiotic and symbiotic cultivation systems initiated from single spores, advanced microscopy, Sanger sequencing of the glomalin gene, and PacBio sequencing of the partial 45S rRNA gene, we show that four strains of R. irregularis produce spores of two distinct morphotypes, one corresponding to the morphotype described in the R. irregularis protologue and the other having the phenotype of R. fasciculatus. The two spore morphs are easily distinguished by spore colour, thickness of the subtending hypha, thickness of the second wall layer, lamination of the innermost layer, and the dextrinoid reaction of the two outer spore wall layers to Melzer's reagent. The glomalin gene of the two spore morphs is identical and that of the PacBio sequences of the partial SSU-ITS-LSU region (2780 bp) obtained from single spores of the R. cf fasciculatus morphotype has a median pairwise similarity of 99.8% (SD = 0.005%) to the rDNA ribotypes of R. irregularis DAOM 197198. Based on these results, we conclude that the model AMF species R. irregularis is dimorphic, which has caused taxonomic confusion in culture collections and possibly in AMF research.

Mode of action of potential biocontrol agents against Fusarium species and Cochliobolus sativus.

The antagonistic activities of three potential biocontrol agents (BCAs), Clonostachys miodochialis, C. rosea, and Minimedusa polyspora, were tested in vitro against the target fungi Fusarium acuminatum, F. avenaceum, F. equiseti, F. graminearum, and Cochliobolus sativus. In vitro dual-culture assay revealed that the BCAs were able to inhibit the mycelial growth of most of the target fungi, with the most common mode of action being mycoparasitism. After 5 d of incubation, depending on the host-parasite interface, biotrophic mycoparasitism was observed via attachment structures on F. avenaceum, F. equiseti, and C. sativus. Haustoria and appressoria were formed by C. miodochialis and M. polyspora on F. avenaceum and C. sativus, respectively. Clamp-like structures were also produced by both BCAs, depending on the host fungus. Clonostachys rosea formed only contact points on its hosts. Coiling structure was observed with C. rosea and M. polyspora occasionally in pure culture but more abundantly in the dual-culture assay. After an additional 3 to 5 d of incubation, the BCAs started damaging their host cells. Asexual fruiting bodies of the BCAs spread necrotrophically on F. acuminatum and F. graminearum and began destroying their mycelia after the initial 5 d of incubation. Furthermore, mycelial dissolution of F. acuminatum by an excreted substance was observed at a distance before direct contact with C. miodochialis. The diffuse metabolite assay revealed that the highest inhibition of the three BCAs was on C. sativus and their least effectiveness was observed on F. graminearum. Overall, these results provide evidence that C. miodochialis, C. rosea, and M. polyspora are potential candidates for biological control of the tested target fungi. This constitutes the first report that these three BCAs are able to establish an initial biotrophic relationship followed by a necrotrophic lifestyle. In addition, C. miodochialis has not previously been reported as a BCA.

Axenic growth of the arbuscular mycorrhizal fungus Rhizophagus irregularis and growth stimulation by coculture with plant growth-promoting rhizobacteria.

Arbuscular mycorrhizal (AM) fungi are ecologically important for the growth and survival of most vascular plants. These fungi are known as obligate biotrophs that acquire carbon solely from host plants. A 13C-labeling experiment revealed the ability of axenically grown Rhizophagus irregularis DAOM 197198 to derive carbon from axenic culture on a relatively novel medium containing two sources of palmitic acid developed by Ishii (designated IH medium). In a separate experiment, this model fungus grew larger mycelia and produced more daughter spores on the IH medium in the presence of two Variovorax paradoxus strains than in axenic culture. In contrast, a strain of Mycobacterium sp. did not influence the growth of the AM fungus. Rhizophagus irregularis produced branched absorbing structures on the IH medium and, in monoxenic culture with V. paradoxus, sometimes formed densely packed hyphal coils. In this study, we report for the first time the formation of coarse terminal pelotons and of terminal and intercalary very fine (≈ 1 µm diameter) hyphal elongations, which could form daughter spores in the presence of V. paradoxus. This study shows the value of IH medium and certain rhizobacteria in the culture of R. irregularis DAOM 197198 in vitro.

Temporal trends in severe morbidity and mortality associated with ectopic pregnancy requiring hospitalisation in Washington State, USA: a population-based study.

OBJECTIVE: To examine temporal trend in maternal mortality/severe morbidity associated with hospitalisation due to ectopic pregnancy. DESIGN: A population-based observational study. SETTING AND PARTICIPANTS: All women hospitalised for ectopic pregnancy in Washington State, USA, 1987-2014 (n=20 418). The main composite outcome of severe morbidity/mortality included death, sepsis, need for transfusion, hysterectomy and systemic or organ failure, identified by diagnostic and procedure codes from hospitalisation files. Severe morbidity/mortality due to ectopic pregnancy were expressed as incidence ratios among women of reproductive age (15-64 years) and among women hospitalised for ectopic pregnancy. Comparisons were made between 1987-1991 (reference) and 2010-2014 using ratios of incidence ratios (RR) and ratio differences (RD). The Cochran-Armitage test for trend assessed statistical significance; logistic regression was used to obtain adjusted OR (AOR) and 95% CI, adjusted for demographic factors and comorbidity. RESULTS: Hospitalisation for ectopic pregnancy declined from 0.89 to 0.16 per 1000 reproductive age women between 1987-1991 and 2010-2014 (p<0.001). Among reproductive age women, ectopic pregnancy mortality remained stable (0.03 per 100 000); and mortality/severe morbidity increased among women aged 25-34 years (p=0.022). Among women hospitalised for ectopic pregnancy, mortality increased from 0.29 to 1.65 per 1000 between 1987-1991 and 2010-2015 (p=0.06); severe morbidity/mortality increased from 3.85% to 19.63% (RR=5.10, 95% CI 4.36 to 5.98; RD=15.78 per 100 women, 95% CI 13.90 to 17.66; AOR for 1-year change was 1.08, 95% CI 1.07 to 1.08). CONCLUSIONS: Hospitalisation for ectopic pregnancy declined in Washington State, USA, between 1987 and 2014; however, mortality/severe morbidity associated with ectopic pregnancy increased in female population aged 25-34 years.

Endophytic hyphal compartmentalization is required for successful symbiotic Ascomycota association with root cells.

Root endophytic fungi are seen as promising alternatives to replace chemical fertilizers and pesticides in sustainable and organic agriculture systems. Fungal endophytes structure formations play key roles in symbiotic intracellular association with plant-roots. To compare the morphologies of Ascomycete endophytic fungi in wheat, we analyzed growth morphologies during endophytic development of hyphae within the cortex of living vs. dead root cells. Confocal laser scanning microscopy (CLSM) was used to characterize fungal cell morphology within lactofuchsin-stained roots. Cell form regularity Ireg and cell growth direction Idir, indexes were used to quantify changes in fungal morphology. Endophyte fungi in living roots had a variable Ireg and Idir values, low colonization abundance and patchy colonization patterns, whereas the same endophyte species in dead (gamma-irradiated) roots had consistent form of cells and mostly grew parallel to the root axis. Knot, coil and vesicle structures dominated in living roots, as putative symbiotic functional organs. Finally, an increased hypha septation in living roots might indicate local specialization within endophytic Ascomycota. Our results suggested that the applied method could be expanded to other septate fungal symbionts (e.g. Basidiomycota). The latter is discussed in light of our results and other recent discoveries.