Mutations across Diverse Domains of CjXDR1 Lead to Multidrug Resistance in Clarireedia jacksonii.

Recently, Clarireedia jacksonii has emerged as a significant pathogen threatening turfgrass, and its escalating resistance to multiple drugs often undermines field interventions. This study highlighted the critical role of the fungus-specific transcription factor CjXDR1 (formerly ShXDR1) in regulating multidrug resistance (MDR) in C. jacksonii. This was demonstrated through experiments involving CjXDR1-knockout and CjXDR1-complemented strains. Our sequence analysis revealed five mutations in CjXDR1: G445D, K453E, S607F, D676H, and V690A. All five gain-of-function (GOF) mutations were confirmed to directly contribute to MDR against three different classes of fungicides (propiconazole: demethylation inhibitor, boscalid: succinate dehydrogenase inhibitor, and iprodione: dicarboximide) using the genetic transformation system and in vitro fungicide-sensitivity assay. Comparative transcriptome analysis revealed that CjXDR1 and its GOF mutations led to the overexpression of downstream genes encoding a Phase I metabolizing enzyme (CYP68) and two Phase III transporters (CjPDR1 and CjAtrD) previously reported. Knockout mutants of CYP68, CjPDR1, CjAtrD, and double-knockout mutants of CjPDR1 and CjAtrD exhibited increased sensitivity to all three fungicides tested. Among these, the CYP68-knockout mutants displayed the highest sensitivity to propiconazole, while the CjPDR1 knockout mutant exhibited significantly increased sensitivity to all three fungicides. Double-knockout mutants of CjPDR1 and CjAtrD displayed greater sensitivity than the single knockouts. In conclusion, multiple GOF mutants in CjXDR1 contribute to MDR by upregulating the expression of CjPDR1, CjAtrD, and CYP68. This study enhances our understanding of the molecular mechanisms underlying MDR in plant pathogenic fungi, providing valuable insights into GOF mutation structures and advancing the development of antifungal drugs.

[Effect of Modified Xiaoyao Powder on intestinal barrier and intestinal flora in mice with metabolic associated fatty liver disease based on "gut-liver axis"].

This study explores the effects and mechanisms of Modified Xiaoyao Powder on the intestinal barrier and intestinal flora in mice with metabolic associated fatty liver disease(MAFLD) based on the " gut-liver axis". Sixty male C57BL/6 mice were randomly divided into the normal group, model group, bifidobacterium tetrad tablet group(SQ), and Modified Xiaoyao Powder groups with low,medium and high doses(XL, XM, XH), with 10 mice in each group. All the mice were administrated with a high-fat diet to build the MAFLD model except the normal group and then treated with related drugs for 12 weeks. Body mass, liver wet weight, and liver index were detected. Serum aspartate aminotransferase(AST), alanine aminotransferase(ALT), total cholesterol(TC), triacylglycerol(TG), low density lipoprotein cholesterol(LDL-C), high density lipoprotein cholesterol(HDL-C), and lipopolysaccharide(LPS)levels were detected using the biochemical kits. The contents of tumor necrosis factor-α(TNF-α) and interleukin(IL-6) in the liver were tested simultaneously. The morphological changes of the liver and intestine were observed using hematoxylin-eosin(HE) staining and oil red O staining. The goblet cells in the ileum were detected by periodic acid Schiff and alcian blue stain(AB-PAS) staining.The expression of zonula occludens-1(ZO-1), recombinant occludin(occludin), and recombinant claudin 1(claudin-1) in ileum and colon were detected by immunohistochemistry and Western blot. The changes of intestinal flora in mice were analyzed by 16S rRNA gene sequencing. The results showed that compared with the normal group, body weight, liver wet weight and liver index in the model group increased. The contents of TC, TG, ALT, AST, LDL-C, and LPS in the serum of the model group increased, while HDL-C decreased. Meanwhile, the contents of TNF-α and IL-6 in liver tissue increased and liver lipid accumulation increased, indicating successful model induction. Compared with the model group, body weight, liver wet weight, and liver index were decreased in XM,XH groups and SQ group. Serum levels of TC, TG, LDL-C, ALT and AST in XM group and SQ group were significantly decreased,and HDL-C levels were increased. The levels of IL-6, TNF-α in liver tissue and serum LPS in the XL, XM groups and SQ group were significantly decreased. The protein expression of claudin-1, occludin and ZO-1 in XL, XM groups and SQ group were increased. The analysis of intestinal flora showed that compared with the model group, Modified Xiaoyao Powder with a medium dose could significantly improve the richness and diversity of intestinal flora in mice. At the phylum level, the Firmicutes/Bacteroidetes(F/B) ratio decreased; at the genus level, Lactobacillus, Brautella, Bacteroides, and Ackermannia increased, while Prevotella, Desulfovibrio and Turicibacter decreased. The main differential species were Odorbacteraceaeae and Peptostreptococcaceae. In conclusion, Modified Xiaoyao Powder could inhibit inflammation, regulate intestinal flora homeostasis, and promote the repair of the intestinal mucosal barrier in mice with MAFLD.

A rapid real-time PCR assay for detecting Microdochium paspali causing sparse leaf patch on seashore paspalum and in environmental samples.

BACKGROUND: Sparse leaf patch (SLP) is one of the most significant diseases affecting seashore paspalum (Paspalum vaginatum Sw.), caused by Microdochium paspali. Fast and accurate detection of this pathogen is crucial for effective disease management. However, conventional culture-based methods are time-consuming and often compromised by the presence of other saprophytic or endophytic fungi. RESULTS: In this study, we developed a real-time fluorescent quantitative (q)PCR method based on the internal transcribed spacer (ITS) region of the ribosomal RNA gene to rapidly detect and quantify M. paspali. The qPCR assay demonstrated the ability to detect all 12 tested isolates of M. paspali, with no cross-reactions observed when tested against 30 isolates of other fungal pathogens from turfgrass samples. The detection limit of the qPCR method was as low as 3.65 × 102 copies µL-1 of M. paspali genomic DNA, and the entire detection process could be completed within 1 h. The fluorescence signal was detectable in the leaf tissues of seashore paspalum without apparent disease symptoms as early as 24 h postinoculation with M. paspali. Moreover, the qPCR method successfully detected M. paspali in both asymptomatic and symptomatic turfgrass samples, including leaf, stem, root and rhizosphere soil, indicating that this assay can significantly enhance the detection of M. paspali. CONCLUSION: The study developed a rapid real-time qPCR assay for the detection of M. paspali causing SLP on seashore paspalum and in environmental samples, which has important implications for early warning and management of SLP. © 2024 Society of Chemical Industry.

Probability of rainstorm and flood disasters due to extreme precipitation in Fen River Basin, China.

Analysis of the probability of extreme precipitation events leading to rainstorm and flood disasters can aid in disaster prevention policy development. Using daily precipitation data from 16 meteorological stations from 1960 to 2019, we calculated eight extreme precipitation indices to analyze the spatio-temporal characteristics of extreme precipitation in the Fen River Basin (FRB) through ensemble empirical mode decomposition and Kriging interpolation. Extreme precipitation events and disasters were defined based on a combination of the antecedent precipitation index (API) and extreme precipitation on the event day and classified; extreme precipitation and the API were ranked from small to large and classified into dry, wet, and moderate (mod) precipitation periods, respectively, yielding nine extreme precipitation event categories. The probability of disasters caused by different types of extreme precipitation events was calculated using a binomial distribution. The results are as follows: (1) between 1960 and 2019, except for extreme precipitation period length, which continuously increased, the extreme precipitation indices changed from a downward to an upward trend since the 1980s. All extreme precipitation indices showed similar interannual variation over short periods and different interdecadal variation over long periods. (2) The extreme precipitation indices showed latitudinal and zonal spatial divergence patterns, but different spatial characteristics were observed around the 1980s. (3) More than 70% of extreme precipitation events in the midstream and downstream fell into four categories: "dry-dry," "dry-mod," "mod-dry," and "mod-mod." (4) A single category VII (VIII) extreme precipitation event in the midstream (downstream) had a maximum probability of causing disaster of 14%. When more than four extreme precipitation events occurred in a year, the probability of one disaster was the highest and that of four or more disasters was < 0.1%. The probability of rainstorm and flood disasters increased gradually with increasing frequency of annual extreme precipitation events.

Temporal Changes of Fish Diversity and Driver Factors in a National Nature Reserve, China.

Freshwater-fish diversity declined rapidly due to multiple anthropogenic disturbances. The loss of fish diversity often manifested itself in taxonomic homogenization over time. Knowledge of multi-faceted diversity (i.e., species, functional, and phylogenetic diversity) perspectives is important for biodiversity assessment and conservation planning. Here, we analyzed the change of the species diversity and phylogenetic diversity of fish in 2008 and 2021 as well as explored the driver factors of the biodiversity patterns in the Lushan National Nature Reserve. The results showed that the species diversity and phylogenetic diversity of fish have declined from 2008 to 2021, with five species lost over time. We found an overall homogenization trend in the fish fauna of the study area, with a 4% increase in taxonomic similarity among the rivers. Additionally, we found that community structure of fish was significantly different among the rivers, and environmental filtering was the main contributor to the phylogenetic diversity of fish in 2008 and 2021. This study provides new insight into the patterns and drivers of fish-biodiversity change in the broader Yangtze River basin and informs management efforts.

The Effects of Water Level Fluctuation on Zooplankton Communities in Shahu Lake Based on DNA Metabarcoding and Morphological Methods.

Background: The water level of Poyang Lake (China) fluctuates seasonally. Shahu Lake, a smaller body of water connected to Poyang Lake during the wet season, is separated in the dry season. Due to a special fishing method termed 'lake enclosed in autumn', the water level is lowered and reaches its lowest point in January, which is <0.5 m deep in the middle of the lake. Our research investigated the effect of water level changes on the zooplankton community composition in Shahu Lake. Methods: We used both DNA metabarcoding method (MBC) (18S rRNA gene V4 region) and morphological method (MOI) to track the zooplankton community structure over four seasons in Shahu Lake (China). Results: Totals of 90 and 98 species of zooplankton were detected by MOI and MBC, respectively, with rotifers being the main zooplankton component. The α-diversity index of both methods increased from spring to summer and decreased from summer to autumn, reaching the lowest value in winter. NMDS and a cluster analysis showed that all zooplankton communities detected by MOI and MBC were significantly separated by season. The zooplankton community in winter was separated from that of the other three seasons, but the summer and autumn communities were more similar. Conclusions: Changes in the water level had significant effects on the zooplankton community composition. We found that MBC was more able to detect the differences in the zooplankton composition than MOI. MBC also had more advantages in copepod recognition. In our study, 37 species of copepods were detected by MBC, but only 11 species were detected by MOI. We concluded that MBC should be used to research the seasonal variations of zooplankton.

Refined spatiotemporal analysis of drought characteristics under different characteristic variable matchings: a case study of the middle reaches of the Yellow River basin, China.

The refined assessment of the spatiotemporal characteristics of droughts is of great significance for drought evaluation. Based on monthly precipitation and temperature grid data (1961-2019) in the middle reaches of the Yellow River basin (MYRB), the standardized precipitation evapotranspiration index (SPEI) was calculated at monthly, seasonal, and annual scales. The run theory was used to extract the drought features at the monthly scale, and the spatiotemporal characteristics of different drought levels were analyzed using Mann-Kendall mutation tests and spatial interpolation. The Moran' I was used to analyze the spatial heterogeneity of droughts. The results showed that the drought trend in the MYRB increased from 1961 to 2019, with the SPEI exhibiting an overall decreasing rate of - 0.1145/decade. Decreasing rates were observed in spring (- 0.1356/decade), summer (- 0.0362/decade), and autumn (- 0.0745/decade), whereas an increasing rate was observed in winter (0.0781/decade). Only extreme droughts were long term, with an intensity as low as - 22.29. The highest frequencies were observed for mild-moderate droughts, which mainly showed high-value clusters in the western and central regions. The frequencies of severe-extreme droughts mainly presented low-value clusters in the northern and southwestern areas. The frequencies of mild and severe droughts exhibited significant spatial cluster characteristics, while the drought intensity showed non-significant spatial clusters and a random distribution. The high and low values of drought intensity were mainly clustered in the middle-upper reaches. The research results provide reference for disaster prevention and mitigation, agricultural planning, and water resource allocation in the MYRB.

Effects of Inulin Propionate Ester on Obesity-Related Metabolic Syndrome and Intestinal Microbial Homeostasis in Diet-Induced Obese Mice.

Short-chain fatty acid (SCFA) plays an important role in improving obesity and related metabolic syndrome induced by high-fat diet. We used the prepared inulin propionate ester (IPE) as a system for the targeted release of propionate to the colon to elucidate the role of IPE in regulating obesity and metabolic syndrome, and intestinal microbial homeostasis, in diet-induced obese mice. With this strategy, IPE significantly increased the SCFA contents in the colon and resulted in significant body weight reduction, insulin resistance amelioration, and gastrointestinal hormone (glucagon-like peptide and peptide YY) secretion (P < 0.05). The IPE intervention reduced liver fatty accumulation, which improved obesity-related fatty liver disease (P < 0.05). IPE supplementation increased the richness and diversity of the microbial community and altered bacterial population at both the phylum and family level. Intestinal microbial results showed that the relative abundance of Desulfovibrionaceae and Erysipelotrichaceae, which promote the production of inflammatory factors, was reduced. Our results demonstrate that IPE can be used as an effective strategy for delivering propionate to obese mice colon, which can ameliorate obesity and associated metabolic syndrome and modify intestinal microbial homeostasis.