An FYVE-Domain-Containing Protein, PsFP1, Is Involved in Vegetative Growth, Oxidative Stress Response and Virulence of Phytophthora sojae.

Proteins that contain the FYVE zinc-finger domain are recruited to PtdIns3P-containing membranes, participating in numerous biological processes such as membrane trafficking, cytoskeletal regulation, and receptor signaling. However, the genome-wide distribution, evolution, and biological functions of FYVE-containing proteins are rarely reported for oomycetes. By genome mining of Phytophthora sojae, two proteins (PsFP1 and PsFP2) with a combination of the FYVE domain and the PX domain (a major phosphoinositide binding module) were found. To clarify the functions of PsFP1 and PsFP2, the CRISPR/Cas9-mediated gene replacement system was used to knock out the two genes respectively. Only heterozygous deletion mutants of PsFP1 were recovered, and the expression level of PsFP1 in the heterozygous knockout transformants was significantly down-regulated. These PsFP1 mutants showed a decrease in mycelial growth and pathogenicity and were more sensitive to hydrogen peroxide. These phenotypes were recovered to the level of wild-type by overexpression PsFP1 gene in the PsFP1 heterozygous knockout transformant. In contrast, deletion of PsFP2 had no significant effect on vegetative growth, asexual and sexual reproduction, pathogenicity, or oxidative stress sensitivity. PsFP1 was primarily localized in vesicle-like structures and both the FYVE and PX domains are important for its localization. Overall, our results indicate that PsFP1 plays an important role in the vegetative growth and virulence of P. sojae.

Characteristics of gut microbiome in patients with pediatric solid tumor.

Background: Pediatric solid tumors are a common malignant disease in children, and more and more studies have proved that there is an inseparable relationship between adult tumors and intestinal microbiome, but the changes in the intestinal microbiota of pediatric solid tumor (PST) patients have been scarcely examined. This study aims to examine the differences in the intestinal microbiota features between patients diagnosed with PST and healthy controls (HCs). Methods: To elucidate the unique characteristics of the gut microbiota in pediatric patients with solid tumors, we recruited 23 PST patients and 20 HCs. A total of 43 stool samples were gathered, and then 16S rRNA sequencing was performed. Results: We noticed a noticeable pattern of elevated diversity in the gut microbiota within the PST groups. The differences in microbial communities among two groups were remarkable, regarding the analysis at the class level, the abundance of Bacilli was markedly increased in PST patients compared to HCs (P < 0.05), regarding the analysis at the genus level, The presence of Enterococcus was significantly higher in PST cases compared to HCs (P < 0.01), while Lachnospiraceae unclassified, Lachnospira, Haemophilus and Colidextribacter in PST cases, the abundance was significantly reduced. (P < 0.05), 6 genera, including Bacilli, Lactobacillales, Enterococcaceae and Morganella, showed a significant enrichment compared to healthy controls, while 10 genera, including Bilophila, Colidextribacter, Pasteurellales, Haemophilus, Lachnospiraceae unclassified, Lachnospira and Fusobacteriales, were significant reduction in the PST groups. Conclusion: Our research conducted the characterization analysis of the gut microbiota in PST patients for the first time. More importantly, there are some notable differences in the gut microbiota between PST patients and healthy controls, which we believe is an interesting finding.

Biological activity and systemic translocation of the new tetrazolyloxime fungicide picarbutrazox against plant-pathogenic oomycetes.

BACKGROUND: Picarbutrazox is a new tetrazolyloxime fungicide discovered in 2014 by Nippon Soda. It is mostly used to protect against Phytophthora spp. and Pythium spp. However, little is known of its inhibition spectrum, protective and curative activity, and systemic translocation in plants. RESULTS: While picarbutrazox did not show obvious antifungal activity, it exhibited significant activity against oomycetes, including Phytophthora spp., Pythium spp. and Phytopythium spp.. The effective concentration for 50% growth inhibition (EC50) values of picarbutrazox against 16 oomycetes ranged from 3.1 × 10-4 and 7.27 × 10-3 µg mL-1. Furthermore, picarbutrazox could markedly inhibited the mycelial development, sporangia production, zoospore release, and cyst germination of Phytophthora capsici, with EC50 values of 1.34 × 10-3, 1.11 × 10-3, 4.85 × 10-3, and 5.88 × 10-2 µg mL-1, respectively. Additionally, under greenhouse conditions, the protective and curative activities of picarbutrazox at 200 mg L-1 (100%, 41.03%) against the P. capsici infection in peppers were higher than those of the reference fungicide dimethomorph at 200 mg L-1 (77.52%, 36.15%). High-performance liquid chromatography analysis confirmed that picarbutrazox showed excellent systemic translocation in pepper plants. CONCLUSION: The results showed that picarbutrazox markedly inhibited the important plant oomycete pathogens including Phytophthora spp., Pythium spp. and Phytopythium spp.. It also displayed excellent protective, curative and systemic translocation activity. Picarbutrazox thus has significant potential for preventing and controlling diseases caused by oomycetes. © 2024 Society of Chemical Industry.

Oral microbiome characteristics in patients with pediatric solid tumor.

Background: Pediatric solid tumor, the abnormal proliferation of solid tissues in children resulting in the formation of tumors, represent a prevailing malignant ailment among the younger population. Extensive literature highlights the inseparable association linking oral microbiome and adult tumors, but due to differences in age of onset, characteristics of onset, etc., there are many differences between Pediatric solid tumors and adult tumors, and therefore, studying the relationship between Pediatric solid tumor and the oral microbiota is also essential. Methods: To unravel the distinct characteristics of the oral microbiota within Pediatric solid tumor patients, 43 saliva samples, encompassing 23 Pediatric solid tumor patients and 20 healthy controls, were diligently procured. A meticulous screening process ensued, and conducted microbial MiSeq sequencing after screening. Results: We documented the oral microbiome attributes among pediatric diagnosed with solid tumors (PST), and meanwhile, we observed a significant trend of decreased oral microbiota diversity in the pediatric solid tumor group. There were notable disparities in microbial communities observed between the two groups, 18 genera including Veillonellaceae, Firmicutes unclassified, Coriobacteriia, Atopobiaceae, Negativicutes, were significantly enriched in PST patients, while 29 genera, including Gammaproteobacteria, Proteobacteria, Burkholderiales, Neisseriaceae, were dominant in the HCs group. It was found that PST group had 16 gene functions, including Amino acid metabolism, Cysteine and methionine metabolism, Photosynthesis antenna proteins, Arginine and proline metabolism, and Aminoacyl tRNA biosynthesi, were significantly dominant, while 29 gene functions that prevailed in HCs. Conclusion: This study characterized the oral microbiota of Pediatric solid tumor patients for the first time, and importantly, targeted biomarkers of oral microbiota may serve as powerful and non-invasive diagnostic tools for pediatric solid tumor patients.

Multiple point mutations in PsORP1 gene conferring different resistance levels to oxathiapiprolin confirmed using CRISPR-Cas9 in Phytophthora sojae.

BACKGROUND: Oxathiapiprolin is among the first commercial oxysterol-binding protein inhibitors (OSBPIs) developed by DuPont Corporation and shows excellent activity against plant-pathogenic oomycetes. Although more than 21 target site mutations have been identified in insensitive oomycetes, only G770V, G839W, and ΔN837 have been verified to confer oxathiapiprolin resistance in Phytophthora capsici or P. sojae. The effect of other mutations on OSBPIs sensitivity requires urgent investigation. RESULTS: P. sojae transformants containing 16 mutations of PsORP1 were recovered using the CRISPR-Cas9 system. Transformants containing L733W, S768F, S768Y, N837Y, N837F, P861H, L863W, or I877Y showed high oxathiapiprolin resistance, with resistant factors (RFs) > 3000. Point mutations S768K, S768I, G770L, G770P, G770A, ΔG818/F819, N837I, and I877F exhibited low resistance, with RFs < 80. Phenotype assays revealed that the most highly resistant transformants showed enhanced or similar pathogenicity, oospore production, and cyst gemination. However, most transformants displayed decreased sporangia and zoospore production compared with parental wild-type P6497. CONCLUSION: This study demonstrated that L733W, S768F, S768Y, N837Y, N837F, P861H, L863W, and I877Y in PsORP1 confer high oxathiapiprolin resistance in P. sojae.

Sensitivity of Pythium spp. and Phytopythium spp. and tolerance mechanism of Pythium spp. to oxathiapiprolin.

BACKGROUND: Oxathiapiprolin, developed by DuPont, is the only commercial oxysterol-binding protein inhibitor (OSBPI) of oomycete pathogens. Although the activity of oxathiapiprolin on some Pythium spp. and Phytopythium spp. has been reported, it has not been tested on many other species, and little is known about the mechanisms of Pythium spp. that are tolerant to it. RESULTS: Oxathiapiprolin exhibited a strong inhibitory effect on mycelial growth of Phy. litorale, Phy. helicoides and Phy. chamaehyphon, with EC50 values ranging from 0.002 to 0.013 µg mL-1 . It also showed good effectiveness against Py. splendens and two Py. ultimum isolates, with EC50 values ranging from 0.167 to 0.706 µg mL-1 , but showed no activity against 14 other Pythium spp. Oxathiapiprolin provoked a slight upregulation of PuORP1 in Py. ultimum, but it did not lead to PaORP1-1 or PaORP1-2 overexpression in Py. aphanidermatum. Transformation and expression of PuORP1, PaORP1-1 or PaORP1-2 in the sensitive wild-type Phytophthora sojae isolate P6497 confirmed that either the PuORP1, PaORP1-1 or PaORP1-2 was responsible for the observed oxathiapiprolin tolerance. CONCLUSION: This study showed that oxathiapiprolin had excellent activity against Phytopythium spp. but displayed a differentiated activity against different Pythium spp. ORP1s in Pythium spp. are positively related to the tolerance of Pythium species to oxathiapiprolin. © 2020 Society of Chemical Industry.