The association of triglyceride-glucose-waist circumference with metabolic associated fatty liver disease and the severity of liver steatosis and fibrosis in American adults: a population-based study.

BACKGROUND: The global obesity pandemic has led to an alarming rise in the prevalence of metabolic-associated fatty liver disease (MAFLD), making it a substantial clinical and economic burden on society. Early detection and effective treatment of MAFLD are imperative to mitigate its impact. METHODS: This cross-sectional study was conducted involving 4634 adults from the National Health and Nutrition Examination Surveys (NHANES) 2017-2018 cycle. Transient elastography (TE) was used to diagnose MAFLD and assess the extent of liver steatosis and fibrosis. Multivariate logistic regression models were utilized to examine the association between the triglyceride and glucose index-waist circumference (TyG-WC) and the risk of MAFLD, liver fibrosis, and steatosis. RESULTS: A positive association between TyG-WC and MAFLD persisted across all three models: model1: OR = 8.44, 95% CI: 6.85-10.38 (unadjusted), model2: OR = 8.28, 95% CI: 6.53-10.50 (partially adjusted), and model3: OR = 7.98, 95% CI: 4.11-15.46 (fully adjusted). Further investigation through interaction and stratified analysis revealed that this association was more pronounced in the non-obese and Non-Hispanic White persons groups. Moreover, a non-linear relationship analysis unveiled threshold and saturation effects between TyG-WC and MAFLD. Specifically, a TyG-WC value of approximately 600 may represent the threshold effect for MAFLD risk, while 1200 may signify the saturation effect of MAFLD risk. Finally, a robust correlation between TyG-WC and the severity of liver steatosis and fibrosis was found. CONCLUSIONS: The findings suggest that the TyG-WC index exhibits excellent predictive value for MAFLD in the general American population.

Molecular mechanism of TRIM32 in antiviral immunity in rainbow trout (Oncorhynchus mykiss).

TRIM family proteins are widely found in multicellular organisms and are involved in a wide range of life activities, and also act as crucial regulators in the antiviral natural immune response. This study aimed to reveal the molecular mechanism of rainbow trout TRIM protein in the anti-IHNV process. The results demonstrated that 99.1% homology between the rainbow trout and the chinook salmon (Oncorhynchus tshawytscha) TRIM32. When rainbow trout were infected with IHNV, the TRIM32 was highly expressed in the gill, spleen, kidney and blood. Meanwhile, rainbow trout TRIM32 has E3 ubiquitin ligase activity and undergoes K29-linked polyubiquitination modifications dependent on the RING structural domain was determined by immunoprecipitation. TRIM32 could interact with the NV protein of IHNV and degrade NV protein through the ubiquitin-proteasome pathway, and was also able to activate NF-κB transcription, thereby inhibiting the replication of IHNV. Moreover, the results of the animal studies showed that the survival rate of rainbow trout overexpressing TRIM32 was 70.2% which was significantly higher than that of the control group, and stimulating the body to produce high levels of IgM when the host was infected with the virus. In addition, TRIM32 can activate the NF-κB signalling pathway and participate in the antiviral natural immune response. The results of this study will help us to understand the molecular mechanism of TRIM protein resistance in rainbow trout, and provide new ideas for disease resistance breeding, vaccine development and immune formulation development in rainbow trout.

CysB Is a Key Regulator of the Antifungal Activity of Burkholderia pyrrocinia JK-SH007.

Burkholderia pyrrocinia JK-SH007 can effectively control poplar canker caused by pathogenic fungi. Its antifungal mechanism remains to be explored. Here, we characterized the functional role of CysB in B. pyrrocinia JK-SH007. This protein was shown to be responsible for the synthesis of cysteine and the siderophore ornibactin, as well as the antifungal activity of B. pyrrocinia JK-SH007. We found that deletion of the cysB gene reduced the antifungal activity and production of the siderophore ornibactin in B. pyrrocinia JK-SH007. However, supplementation with cysteine largely restored these two abilities in the mutant. Further global transcriptome analysis demonstrated that the amino acid metabolic pathway was significantly affected and that some sRNAs were significantly upregulated and targeted the iron-sulfur metabolic pathway by TargetRNA2 prediction. Therefore, we suggest that, in B. pyrrocinia JK-SH007, CysB can regulate the expression of genes related to Fe-S clusters in the iron-sulfur metabolic pathway to affect the antifungal activity of B. pyrrocinia JK-SH007. These findings provide new insights into the various biological functions regulated by CysB in B. pyrrocinia JK-SH007 and the relationship between iron-sulfur metabolic pathways and fungal inhibitory substances. Additionally, they lay the foundation for further investigation of the main antagonistic substances of B. pyrrocinia JK-SH007.

Genome-wide association analysis reveals a novel QTL CsPC1 for pericarp color in cucumber.

BACKGROUND: Cucumber is an important melon crop in the world, with different pericarp colors. However, the candidate genes and the underlying genetic mechanism for such an important trait in cucumber are unknown. In this study, a locus controlling pericarp color was found on chromosome 3 of cucumber genome. RESULTS: In this study, the light green inbred line G35 and the dark green inbred line Q51 were crossed to produce one F2 population. Consequently, we identified a major locus CsPC1 (Pericarp color 1). Next, we mapped the CsPC1 locus to a 94-kb region chromosome 3 which contains 15 genes. Among these genes, Csa3G912920, which encodes a GATA transcription factor, was expressed at a higher level in the pericarp of the NIL-1334 line (with light-green pericarp) than in that of the NIL-1325 line (with dark-green pericarp). This study provides a new allele for the improvement of cucumber pericarp color. CONCLUSION: A major QTL that controls pericarp color in cucumber, CsPC1, was identified in a 94-kb region that harbors the strong candidate gene CsGATA1.

Fine mapping of a novel QTL CsFSG1 for fruit skin gloss in cucumber (Cucumis sativus L.).

Cucumber (Cucumis sativus L.) is an important vegetable crop that is popular with many people. Peel gloss is a highly valued external quality trait that affects the market value of cucumbers, and it directly influences the purchasing psychology of consumers. Nonetheless, the candidate genes and underlying genetic mechanism for this important cucumber trait are not well understood. In this study, we successfully mapped a fruit skin gloss QTL interval to chromosome 3 (26.04-26.14 Mb) using BSA and GWAS methods. Among the eleven candidate genes in the interval, the cytochrome P450 family gene CsCYP86B1 was identified as the candidate for control of fruit skin gloss in cucumber. The expression of CsCYP86B1 in 0-day fruit skin was significantly lower in the low-gloss isogenic line NIL-1334 than in the high-gloss isogenic line NIL-1325. Our findings provide new insights for improving fruit skin gloss in cucumber breeding. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-022-01291-y.

Association of glycated hemoglobin with non-alcoholic fatty liver disease patients and the severity of liver steatosis and fibrosis measured by transient elastography in adults without diabetes.

BACKGROUND: Type 2 diabetes mellitus (T2DM) is a well-known independent risk factor for non-alcoholic fatty liver disease (NAFLD). However, research exploring the association between blood glucose management and the risk of NAFLD status in subjects without diabetes was insufficient. This study aimed to explore the association of glycated hemoglobin (HbA1c) with NAFLD status and the severity of liver steatosis and fibrosis in non-diabetic people. METHODS: A cross-sectional analysis was conducted on 2998 non-diabetic American adults using data from the National Health and Nutrition Examination Survey (NHANES) 2017-2018 cycle. We used multivariable logistic regression models to evaluate the association between HbA1c and NAFLD status and the severity of liver steatosis and fibrosis. Interaction and stratified analyses were additionally performed. RESULTS: The multivariate regression analyses showed that HbA1c was associated independently with NAFLD status in all the models (model1: OR = 2.834, 95%CI: 2.321, 3.461; model 2: OR = 2.900, 95%CI: 2.312, 3.637 and model 3: OR = 1.664, 95%CI: 1.284, 2.156). We further performed the interaction and stratified analyses and discovered a significant interaction between HbA1c and BMI (Pinteraction < 0.05). Finally, a robust link was shown between HbA1c level and the severity of liver steatosis, which was mainly significant in the prediabetes group, while the correlation was not significant in HbA1c level and severity of liver fibrosis after controlling for all the potential confounders. CONCLUSIONS: We concluded that HbA1c level was positively correlated to the risk of developing NAFLD in a large non-diabetic American population. Moreover, HbA1c level was associated with the severity of liver steatosis in subjects with prediabetes, suggesting that routine screening for HbA1c among individuals with prediabetes is necessary.

Inhibitory Effects of Phenazine Compounds and Volatile Organic Compounds Produced by Pseudomonas aurantiaca ST-TJ4 Against Phytophthora cinnamomi.

Phytophthora cinnamomi is an important plant pathogen that is widely distributed worldwide and has caused serious ecological damage and significant economic losses in forests and plantations in many countries. The use of plant growth-promoting rhizobacteria is an effective and environmentally friendly strategy for controlling diseases caused by P. cinnamomi. In this study, we investigated the antagonistic mechanism of Pseudomonas aurantiaca ST-TJ4 against P. cinnamomi through different antagonistic approaches, observations of mycelial morphology, study of mycelial metabolism, and identification of antagonistic substances. The results showed that Pseudomonas aurantiaca ST-TJ4 was able to significantly inhibit mycelial growth, causing mycelial deformation and disrupting internal cell structures. Additionally, pathogen cell membranes were damaged by ST-TJ4, and mycelial cell content synthesis was disrupted. Ultraperformance liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry analyses showed that phenazine compounds and 2-undecanone were the main antagonistic components. The ammonia produced by the ST-TJ4 strain also contributed to the inhibition of the growth of P. cinnamomi. In conclusion, our results confirm that Pseudomonas aurantiaca ST-TJ4 can inhibit P. cinnamomi through multiple mechanisms and can be used as a biological control agent for various plant diseases caused by P. cinnamomi.

Improvement of Sphaeropsis Shoot Blight Disease Resistance by Applying the Ectomycorrhizal Fungus Hymenochaete sp. Rl and Mycorrhizal Helper Bacterium Bacillus pumilus HR10 to Pinus thunbergii.

Ectomycorrhizal fungi (EMFs) form symbioses with plant roots to promote nutrient uptake by plants but it is controversial as to whether they induce disease resistance in plants. Here, we inoculated pine seedlings with Sphaeropsis sapinea, which was presymbiotic with the EMF Hymenochaete sp. Rl, and the mycorrhizal helper bacterium (MHB) Bacillus pumilus HR10, which promotes the formation of Pinus thunbergia-Hymenochaete sp. Rl mycorrhizae. The results showed that inoculation with Hymenochaete sp. Rl, B. pumilus HR10, and the consortium significantly reduced pine shoot blight disease caused by S. sapinea. After inoculation with pathogenic fungi, callose deposition was significantly increased in needles of pine seedlings inoculated with Hymenochaete sp. Rl, B. pumilus HR10, and the consortium, together with an increase in enzymatic and nonenzymatic systemic antioxidant activity as well as early priming for upregulated expression of PR3 and PR5 genes. Our findings suggest that ectomycorrhizal colonization enhances the resistance of pine seedlings to Sphaeropsis shoot blight by triggering a systemic defense response and that interactions between EMFs and MHBs are essential for mycorrhizal-induced disease resistance.

First Report of Botryosphaeria dothidea Causing Leaf Blight on Cornus officinalis in Jiangsu Province, China.

Cornus officinalis Sieb. et Zucc., belonging to the family Cornaceae, is often used as an ornamental plant and is widely distributed in Shandong, Jiangsu, and Zhejiang provinces and other places in China. Since 2020, a new disease with high incidence has been found in Xuanwu Lake Park (32°04'34.53″N 118°48'42.06″E) in Nanjing, Jiangsu Province, China. The symptoms began as small brown lesions formed along the leaf tips, which gradually expanded and became dark brown with a light brown border. A survey of C. officinalis trees in Xuanwu Lake Park showed that approximately 90% of thirty trees were infected, which decreased the ornamental value of C. officinalis. Pieces of leaf tissue (3 to 4 mm²) from the lesion margins were surface sterilized with 75% ethanol for 30 s and 1% NaClO for 90 s. Subsequently, the tissues were rinsed with sterile H2O, placed on potato dextrose agar (PDA) medium and incubated at 25℃ for 5 days. The same fungus was isolated in 90% of the tissues. Pure cultures were obtained by monosporic isolation. A representative isolate, SZY 2-2, was used for morphological and molecular characterization. The colonies were initially white, gradually turning gray green to black with copious gray aerial mycelium after 1 week in culture. Conidia were one-celled, hyaline, smooth, and fusoid to ellipsoid. Conidia measurements were 23.6±1.9×7.2±0.56 µm (n = 50). The morphology of SZY 2-2 matched the description of Botryosphaeria dothidea (Slippers et al. 2004). To verify species identity, the partial sequences of the internal transcribed spacer (ITS) region, translation elongation factor 1 alpha (EF1-a) gene, and beta-tubulin gene (TUB), were amplified from isolate SZY 2-2 with primers ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Koho 1999), and ßt2a/ßt2b (Glass and Donaldson 1995), respectively. The sequences were deposited in GenBank (ON171471 for ITS, ON185540 for EF1-a, and ON185541 for TUB). A BLAST search of GenBank showed that ITS, EF1-a and TUB sequences of SZY 2-2 were similar to those of B. dothidea MN633360 (identity=517/517 bp; 100%), MK783294 (identity=299/299 bp; 100%), and KF005081 (identity=461/461 bp; 100%), respectively. The morphological and molecular results identified the isolate as B. dothidea (Zhai et al. 2014). To fulfill Koch's postulates, a pathogenicity test was conducted using three C. officinalis plants. Five leaves from each tree were wounded and inoculated with mycelial plugs (about 4 mm in diameter) of B. dothidea from a 5-day-old culture grown on PDA, and inoculation with sterile PDA plugs on different leaves of the same tree served as negative controls. The leaves were enclosed in plastic bag along with the branches with a wet cotton ball inside. Sterile H2O2 was sprayed into the plastic bags to keep moisture conditions.Five days later, all inoculated points showed lesions similar to those previously observed in the field, whereas controls were asymptomatic. The pathogen was successfully reisolated from the inoculated symptomatic parts on PDA and had morphology as characterized before, thus fulfilling Koch's postulates. B. dothidea is known as a ubiquitous fungus and operates as both an endophyte and an opportunistic pathogen of trees (Slippers and Wingfield 2007, Zhao et al 2020). Stress factors that predispose trees to disease expression by B. dothidea include drought, defoliation (Theodore et al. 1997), competition, and physical damage (Slippers and Wingfield 2007). This is consistent with the occurrence of the disease in September and association of B. dothidea with the presence of wounds. More investigation is needed to determine the relationship between possible endophytic growth of B. dothidea on C. officinalis and the leaf blight found in Jiangsu Province.

Identification of Two Fungal Pathogens Responsible for Liriodendron chinense × tulipifera Black Spot and Screening of Trichoderma sp. for Disease Control.

Liriodendron chinense × tulipifera black spot is a newly discovered disease that causes yellowing and early shedding of leaves, affecting the growth of Liriodendron trees, and significantly reducing their ornamental value as a garden species. The pathogen responsible for this disease, and how it can be prevented and controlled, are not clear. In this study, the occurrence of this disease was first investigated according to Koch's postulates, and the primary pathogens causing Liriodendron black spot were determined to be Colletotrichum gloeosporioides and Alternaria alternata. Biocontrol strains antagonistic to these two pathogens were then screened from the leaf microorganisms of L. chinense × tulipifera, and a preliminary investigation of the biological control of Liriodendron black spot was performed. Through the screening of antagonistic microorganisms on the leaf surface of L. chinense × tulipifera, the strain Trichoderma koningiopsis T2, which displayed strong antagonism against C. gloeosporioides and A. alternata, was obtained. The T2 strain could inhibit the growth of the two pathogens via three mechanisms: hyperparasitism, volatile and nonvolatile metabolite production, and environmental acidification. The biocontrol experiments in the greenhouse and field showed that initial spraying with a T. koningiopsis T2 spore suspension followed by the two pathogens resulted in the lowest disease incidence. These results confirmed the black spot pathogens of L. chinense × tulipifera, clarified the antagonistic mechanism of T. koningiopsis T2 against the two pathogens, and provided a theoretical basis and technical support for the biological control of the disease.

First Report of Diaporthe eres Causing Leaf Spot on Pseudocydonia sinensis in China.

Pseudocydonia sinensis is a Chinese ornamental plant with great landscaping value. Its fruit is additionally used for medicinal purposes (Lim 2012). In June 2020, a leaf spot disease was observed in the campus of Nanjing Forestry University (32°04'34.53″N 118°48'42.06″E). The symptoms began with irregular red-brown spots, which gradually enlarged, extended and overlapped, with an incidence of 85% (29/34 trees). Pieces of leaf tissue (3 to 4 mm²) from the lesion margins were surface-sterilized with 75% ethanol for 30 s and 1% NaClO for 90 s. Subsequently, the tissues were rinsed with sterile H2O, placed on potato dextrose agar (PDA) medium and incubated at 25℃ for 5 days. The same fungus was isolated from 90% of tissues. Pure cultures were obtained by monosporic isolation.The representative isolate NJMG 5-7 was used for morphological and molecular characterization. The growing fungal colony on PDA was initially white, but gradually turned grey. Pycnidia formation was observed on PDA supplemented with alfalfa stems. The pycnidia produced two different types of conidia, α and ß, which ooze out in yellow creamy mucilaginous masses. Conidiophores were hyaline, cylindrical and smooth, 16.8 to 33.1 × 1.5 to 2.6 µm (n=30). Conidiogenous cells were 13.6 to 29.3 × 1.5 to 2.7 µm (n=30). The α-conidia were, unicellular, hyaline elliptical or fusiform, bi-guttulate, 6.5 to 9.2 × 1.8 to 3.3 µm (n = 50). The ß-conidia were hyaline, aseptate, without guttules, filiform, curved, with obtuse ends, 12.5 to 25 × 1.0 to 1.8 µm (n = 50). To verify species identity, the partial sequences of the internal transcribed spacer (ITS) region, and calmodulin (CAL), translation elongation factor 1 alpha (EF1-a), and beta-tubulin genes (TUB) were amplified from isolate NJMG 5-7 with primers ITS1/ITS4 (White et al. 1990), CAL-228F/CAL-737R (Carbone & Kohn 1999), EF1-728F/EF1-986R (Carbone and Kohn 1999), and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. The sequences were deposited in GenBank (OP223050 for ITS, OP252809 for CAL, OP252807 for EF1-a, and OP252808 for TUB). A BLAST search of GenBank showed that ITS, CAL, EF1-a and TUB sequences of NJMG 5-7 were similar to those of D. eres CBS 138594 (99% identity), AR5193 (99%), AR5193 (99%) and MG281193 (100%), respectively. The morphological and molecular results identified the isolate as D. eres (Feng et al. 2015). To fulfill Koch's postulates, a pathogenicity test was conducted using three P. sinensis plants. Six leaves from each tree were wounded and inoculated with mycelial plugs (about 4 mm in diameter) of D. eres from a 3-day-old culture grown on PDA. Inoculations with sterile PDA plugs on different leaves of the same tree were used as controls. All inoculated leaves were enclosed in plastic bags together with a wet cotton ball inside. Sterile H2O was sprayed into the plastic bags to keep moisture conditions. Five days later, all inoculated points showed lesions similar to those previously observed in the field, whereas controls were asymptomatic. The pathogen was successfully reisolated from the inoculated symptomatic parts on PDA and identified from its morphology, thus fulfilling Koch's postulates. This fungus can cause a variety of diseases. To our knowledge, this is the first report of D. eres causing leaf spots on P. sinensis in the world. These findings provide a foundation for future studies on the epidemiology and control of this newly emerging disease.

Physiological and Transcriptome Analyses Revealed the Mechanism by Which Deferoxamine Promotes Iron Absorption in Cinnamomum camphora.

Iron deficiency causes chlorosis and growth inhibition in Cinnamomum camphora, an important landscaping tree species. Siderophores produced by plant growth-promoting rhizobacteria have been widely reported to play an indispensable role in plant iron nutrition. However, little to date has been determined about how microbial siderophores promote plant iron absorption. In this study, multidisciplinary approaches, including physiological, biochemical and transcriptome methods, were used to investigate the role of deferoxamine (DFO) in regulating Fe availability in C. camphora seedlings. Our results showed that DFO supplementation significantly increased the Fe2+ content, SPAD value and ferric-chelate reductase (FCR) activity in plants, suggesting its beneficial effect under Fe deficiency. This DFO-driven amelioration of Fe deficiency was further supported by the improvement of photosynthesis. Intriguingly, DFO treatment activated the metabolic pathway of glutathione (GSH) synthesis, and exogenous spraying reduced glutathione and also alleviated chlorosis in C. camphora. In addition, the expression of some Fe acquisition and transport-related genes, including CcbHLH, CcFRO6, CcIRT2, CcNramp5, CcOPT3 and CcVIT4, was significantly upregulated by DFO treatment. Collectively, our data demonstrated an effective, economical and feasible organic iron-complexing agent for iron-deficient camphor trees and provided new insights into the mechanism by which siderophores promote iron absorption in plants.

First Report of leaf spot disease caused by Colletotrichum gloeosporioides on Chaenomeles sinensis in China.

Chaenomeles sinensis is a shrub or small arbor of the genus Chaenomeles in Rosaceae, which is widely planted in China. It is a kind of garden ornamental plant and has high economic value. Since 2020, a leaf disease occurred on the foliage of C. sinensis at the campus of Nanjing Forestry University, Nanjing, China. After investigating, C. sinensis was found with leaf spot disease at a 100% infection rate, which causing gigantic ornamental loss. Leaf spots are round to irregular distributing on the leaves, in addition, the color of spots is brown. There are yellow halos on the edge of the lesion. Small leaf tissues (3 to 4 mm2) from lesion margins were surface sterilized with 75% ethanol for 30s and then rinsed with sterile dH2O for three times. Afterwards, placed on potato dextrose agar (PDA) at 25°C. Pure cultures were obtained by monosporic isolation, and a representative isolate (NJTJ.1) was obtained. When cultured on PDA, the colony of NJTJ.1 was white and cottony. On the reverse side, the color of colony nearly light yellow. The colony were placed in the liquid Carboxymethyl cellulose (CMC) medium. After culturing for 24h in a shaker at 25℃ and 150rmp/min, the spore liquid was taken by us. The conidia were one-celled, straight, hyaline, subcylindrical with rounded ends and measured 15.1 to 23.6× 5.4 to 7.9 µm (n =30). Appressoria were one-celled, brown, thick-walled, ellipsoidal, and measured 7.7 to 13.8 × 6.4 to 10.3 µm (n =30). The morphological characteristics of NJTJ.1 fitted with the description of the Colletotrichhum gloeosporioides complex (Weir et al., 2012). For accurate identification, the internal transcribed spacer (ITS), and the genes encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT) and chitin synthase (CHS-1) were amplified with primers ITS1/ITS4, GDF/GDR, ACT-512F/ACT-783R, and CHS-79F/CHS-345R (Zhu et al, 2019), respectively. The sequences were deposited in GenBank [Accession Nos.MT984264, MW030495 and MW030496 to MW030497 for NJTJ.1]. A Blast search of GenBank showed that ITS, GAPDH, ACT and CHS-1 sequences of NJTJ.1 were 99%, 99%, 100% and 100% identical to those of C. gloeosporioides (MH571757.1 ,KY995355.1 , MN058143.1 and MN313581.1). A neighbor-joining phylogenetic tree was generated by combining all sequenced loci in MEGA7. The isolate NJTJ.1 clustered in the C. gloeosporioides clade with 99% bootstrap support. The pathogenicity of the NJTJ.1 was verified both on detached and living leaves. The detached leaves were inoculated with 5-mm mycelial plugs cut from the edge of 6-day old cultures on PDA and 20 µL of spore suspension (106 conidia/mL) and each treatment had 5 replicates. Controls were treated with sterile dH2O. The inocula were placed at a distance of 2 to 3 cm on the leaves which were wounded with a sterile needle. All of them were placed in 20-cm dishes on wet filter paper at 25°C. After 5 days, all the inoculated points showed lesions which were similar to those outdoor observed. Whereas, controls were asymptomatic.At the same time, the plugs of C. gloeosporioides were inoculated on living leaves.After 7 days, the leaves which were inoculated also appeared lesions. This is the first report of C. gloeosporioides causing leaf blotch on Chaenomeles sinensis in China. These data will help develop effective strategies for managing this disease.

Salt Tolerance Mechanism and Species Identification of the Plant Rhizosphere Bacterium JYZ-SD2.

A salt-tolerant microbe strain JYZ-SD2 was investigated to develop biological soil amendments to stimulate salix growth and acclimation in costal salt-affected soils. The salt tolerance mechanism of strain JYZ-SD2 was investigated by detecting the salt-tolerant growth characteristics, biofilm formation, ion distribution, secondary metabolites, and zymogram profiling. The strain was identified by physiological and biochemical characteristics (Biolog), 16S rDNA sequencing, and cry1/7/9 gene expressing. With increasing of NaCl concentration, strain JYZ-SD2 adapted to the increased osmotic pressure by prolonging the retardation period, slowing down the growth rate of the logarithmic phase, increasing spo0A gene expression, increasing biofilm formation, reducing Na+ uptake, and changing the expression of metabolites and intracellular soluble proteins. The results showed that strain JYZ-SD2 could be assigned to Bacillus cereus.

First report of leaf spot disease caused by Neopestalotiopsis chrysea on Carya illinoinensis in China.

Pecan (Carya illinoinensis) is important and widely planted nut tree species in Anhui province in China. In a pecan orchard in Anhui, China, 54% of the 1-year-old container-grown seedlings in the nursery developed leaf spots in September 2019. Initially, the brown spots appeared on the leaves. Later, the spots expanded to become brown circles surrounded by a dark brown border. Under severe infection, defoliation occurred and black acervuli were observed on symptomatic leaves. Disease symptoms were not observed on the fruits. To isolate the pathogen, leaf sections (3 to 4 mm) were excised from the margin of the diseased leaf tissues, surface sterilized in 75% alcohol for 30 s and then in 0.1% mercuric chloride for 30s, rinsed three times in sterilized distilled water, plated on potato dextrose agar (PDA) and incubated at 25 °C in the darkness. Pure cultures were obtained by monosporic isolation. The colony of a representative isolate, CZ-4, growing on PDA was circular, white, and cottony, and the surface undulate and pale luteous on PDA. The reverse similar in color. The conidial masses were black and appeared over PDA plates after 14 days. Conidia (15.41-29.48×4.15-7.54µm) (n=50) were fusiform to ellipsoid and four-septate (one basal and one apical cell hyaline, and three brown median cells), with two to three apical appendages. According to colony and conidia morphology, the isolates were identified as Neopestalotiopsis sp. (Maharachchikumbura et al. 2014). Genomic DNA was extracted from single conidial cultures of a representative isolate CZ-4, and the internal transcribed spacer (ITS), translation elongation factor 1-alpha (TEF1), ß-tubulin (TUB) genes were amplified with the primers described by Wang et al. (2019). The obtained sequences showed 95-99% similarity with those from Neopestalotiopsis chrysea accessions in GenBank. The sequences from this isolate were deposited in GenBank under the following accession numbers: ITS, MT459336; TEF1, MT469880; TUB, MT469881. A neighbor-joining phylogenetic tree was generated by combining all sequenced loci in MEGA7. The isolate CZ-4 clustered in the N. chrysea clade with 98% bootstrap support. To test pathogenicity, ten detached healthy leaves and ten one-year-old Carya illinoinensis plants were inoculated with the same pathogens by spraying 50µl of a conidial suspension (1×106 conidia/ml) on both sides of leaves.As a control treatment, ten additional detached leaves and potted seedlings were inoculated with 50µl sterile water. All plants were covered with clear polyethylene bags and incubated in a greenhouse (Center of Co-Innovation, Institute of Forestry) at 23 ± 5 °C, 80% relative humidity, and a 12-h light/dark cycle.The experiment was repeated three times. Seven days after inoculation, the symptoms were similar to those on the original infected plants, whereas the control leaves remained symptomless. N. chrysea was re-isolated from the lesions, morphologically identified, confirming Koch's postulates. To our knowledge, this is the first report of N. chrysea associated with leaf spot disease on C. illinoinensis. This study provides the foundation to further investigate the biology, epidemiology, and management of this disease.

Volatiles emitted by Pseudomonas aurantiaca ST-TJ4 trigger systemic plant resistance to Verticillium dahliae.

Verticillium dahliae is among the most devastating fungal pathogens, causing significant economic harm to agriculture and forestry. To address this problem, researchers have focused on eliciting systemic resistance in host plants through utilizing volatile organic compounds (VOCs) produced by biological control agents. Herein, we meticulously measured the quantity of V. dahliae pathogens in plants via RTqPCR, as well as the levels of defensive enzymes and pathogenesis-related (PR) proteins within plants. Finally, the efficacy of VOCs in controlling Verticillium wilt in cotton was evaluated. Following treatment with Pseudomonas aurantiaca ST-TJ4, the expression of specific VdEF1-α genes in cotton decreased significantly. The incidence and disease indices also decreased following VOC treatment. In cotton, the salicylic acid (SA) signal was strongly activated 24 h posttreatment; then, hydrogen peroxide (H2O2) levels increased at 48 h, and peroxidase (POD) and catalase (CAT) activities increased to varying degrees at different time points. The malondialdehyde (MDA) content and electrolyte leakage in cotton treated with VOCs were lower than those in the control group, and the expression levels of chitinase (CHI) and PR genes (PR10 and PR17), increased at various time points under the ST-TJ4 treatment. The activity of phenylalanine ammonia lyase (PAL) enzymes in cotton treated with VOCs was approximately 1.26 times greater than that in control plants at 24 h，while the contents of phenols and flavonoids increased significantly in the later stage. Additionally, 2-undecanone and 1-nonanol can induce a response in plants that enhances disease resistance. Collectively, these findings strongly suggest that VOCs from ST-TJ4 act as elicitors of plant defence and are valuable natural products for controlling Verticillium wilt.

High serum klotho levels are inversely associated with the risk of low muscle mass in middle-aged adults: results from a cross-sectional study.

Objective: Muscle mass gradually declines with advancing age, and as an anti-aging protein, klotho may be associated with muscle mass. This study aims to explore the relationship between klotho levels and muscle mass in the middle-aged population. Methods: Utilizing data from the National Health and Nutrition Examination Survey (NHANES) spanning 2011 to 2018, we conducted a cross-sectional analysis on a cohort of individuals aged 40-59. Weighted multivariable analysis was employed to assess the correlation between klotho and low muscle mass, with stratified and Restricted Cubic Spline (RCS) analyses. Results: The cross-sectional investigation revealed a significant negative correlation between klotho levels and the risk of low muscle mass (Model 3: OR = 0.807, 95% CI: 0.712-0.915). A notable interaction between klotho and sex was observed, with a significant interaction effect (P for interaction = 0.01). The risk association was notably higher in females. The risk association was notably higher in females. Additionally, RCS analysis unveiled a significant linear relationship between klotho and low muscle mass (P for nonlinear = 0.9495, P for overall<0.0001). Conclusion: Our observational analysis revealed a noteworthy inverse relationship between klotho and low muscle mass, particularly prominent among female participants. This discovery provides crucial insights for the development of more effective intervention strategies and offers a new direction for enhancing muscle quality in the middle-aged population.

Higher ultra processed foods intake is associated with low muscle mass in young to middle-aged adults: a cross-sectional NHANES study.

Design: Ultra-processed foods (UPFs) have become a pressing global health concern, prompting investigations into their potential association with low muscle mass in adults. Methods: This cross-sectional study analyzed data from 10,255 adults aged 20-59 years who participated in the National Health and Nutritional Examination Survey (NHANES) during cycles spanning from 2011 to 2018. The primary outcome, low muscle mass, was assessed using the Foundation for the National Institutes of Health (FNIH) definition, employing restricted cubic splines and weighted multivariate regression for analysis. Sensitivity analysis incorporated three other prevalent definitions to explore optimal cut points for muscle quality in the context of sarcopenia. Results: The weighted prevalence of low muscle mass was 7.65%. Comparing the percentage of UPFs calories intake between individuals with normal and low muscle mass, the values were found to be similar (55.70 vs. 54.62%). Significantly linear associations were observed between UPFs consumption and low muscle mass (P for non-linear = 0.7915, P for total = 0.0117). Upon full adjustment for potential confounding factors, participants with the highest UPFs intake exhibited a 60% increased risk of low muscle mass (OR = 1.60, 95% CI: 1.13 to 2.26, P for trend = 0.003) and a decrease in ALM/BMI (ß = -0.0176, 95% CI: -0.0274 to -0.0077, P for trend = 0.003). Sensitivity analysis confirmed the consistency of these associations, except for the International Working Group on Sarcopenia (IWGS) definition, where the observed association between the highest quartiles of UPFs (%Kcal) and low muscle mass did not attain statistical significance (OR = 1.35, 95% CI: 0.97 to 1.87, P for trend = 0.082). Conclusion: Our study underscores a significant linear association between higher UPFs consumption and an elevated risk of low muscle mass in adults. These findings emphasize the potential adverse impact of UPFs on muscle health and emphasize the need to address UPFs consumption as a modifiable risk factor in the context of sarcopenia.

Clinical application of nanopore-targeted sequencing technology in bronchoalveolar lavage fluid from patients with pulmonary infections.

The rapid and effective identification of pathogens in patients with pulmonary infections has posed a persistent challenge in medicine, with conventional microbiological tests (CMTs) proving time-consuming and less sensitive, hindering early diagnosis of respiratory infections. While there has been some research on the clinical performance of targeted sequencing technologies, limited focus has been directed toward bronchoalveolar lavage fluid (BALF). This study primarily evaluates the pathogen detection capabilities of nanopore-targeted sequencing (NTS) in BALF, providing a comprehensive analysis. The retrospective study, spanning from January 2022 to November 2023, includes 223 patients exclusively sourced from a single center. We conducted a detailed comparative analysis among NTS, targeted next-generation sequencing (tNGS), and CMTs. Initially, we compared the detection capabilities of NTS and tNGS and found no significant differences in their sensitivity and specificity. Specifically, we observed that the sensitivity of NTS was significantly higher than that of CMTs (74.83% vs 33.11%, P < 0.001). Furthermore, NTS exhibited a higher positivity rate in common pulmonary infections (62.88% vs. 23.48%) and in clinically suspected tuberculosis patients compared to CMTs (87.18% vs. 48.72%). Additionally, NTS showed less susceptibility to antibiotic interference, indicating a more sensitive detection capability, especially in detecting fastidious organisms. It complements GeneXpert in tuberculosis diagnosis and offers excellent advantages in identifying pathogens challenging for CMTs, such as non-tuberculous mycobacteria and viruses. Moreover, NTS significantly shortens the reporting time and is only a quarter of the cost of metagenomic next-generation sequencing. Clearly, NTS can facilitate faster and more cost-effective early diagnosis of respiratory infections.IMPORTANCEThis study holds paramount significance in advancing the field of respiratory infection diagnostics. By assessing the pathogen detection capabilities in bronchoalveolar lavage fluid (BALF) of patients with pulmonary infections, we illuminate the promising potential of nanopore-targeted sequencing (NTS). The findings underscore NTS as a comparable yet distinct alternative to traditional methods like comprehensive conventional microbiological tests (CMTs). Notably, NTS demonstrates a pivotal edge, expanding the spectrum of identified pathogens, particularly excelling in the detection of challenging entities like non-tuberculous mycobacteria and viruses. The study also highlights the complementary role of NTS alongside GeneXpert in the identification of tuberculosis, providing a comprehensive overview of the diagnostic landscape for respiratory infections. This insight carries significant implications for clinicians seeking rapid, cost-effective, and accurate diagnostic tools in the realm of pulmonary infections.

Case Report: A rare case of non-small cell lung cancer with STRN-ALK fusion in a patient in very poor condition treated with first-line ensartinib.

Several cases of STRN-ALK fusion have been reported, and some anaplastic lymphoma kinase (ALK) inhibitors have been shown to be effective for treatment. Nevertheless, no cases of COVID-19 leading to heart failure and respiratory failure have been reported in people older than 70 years treated with ALK inhibitors. The present case report describes a 70-year-old patient with usual chronic obstructive pulmonary disease, diabetes, depression, and carotid plaque disease. Next-generation sequencing of tissue obtained by puncture biopsy revealed a STRN-ALK mutation accompanied by a TP53 mutation. The patient was treated with ensartinib and developed COVID-19 leading to heart failure and respiratory failure; nevertheless, he had a good clinical outcome and exhibited high treatment tolerability.