First Report of Leaf Spot Disease Associated with Faba Bean (Vicia faba) Caused by Pestalotiopsis rosea in China.

Faba bean (Vicia faba L.) is a cool-season legume crop, planted worldwide as an essential source of protein-rich foods, vegetables, and animal feeds. In China, the total cultivated area of faba bean in 2019 was 839,618 square hectometers (hm2) and the production was 1,740,945 tons (t) (Ji et al. 2022). In May 2021, a leaf spot disease on faba bean plants with about 80% disease incidence was observed in a 0.3 ha commercial field located at Zunyi City (27°31'43.80″ N, 106°23'34.27″ E), Guizhou Province, China. The leaves of the early affected plants appeared circular dark brown spots, which then rapidly develop into large irregular shaped lesions if conditions remain favorable. Severe infection can result in extensive defoliation of plants and lesions on pods. Symptomatic leaves were collected and cut into small pieces, surface sterilized with 75% ethanol for 30 s followed by 2% NaClO for 1 min, rinsed with sterile distilled water three times, and incubated on PDA plates amended with streptomycin sulfate (0.5 mg/L) at 25°C for 2-4 days. Two purified cultures were obtained through single-spore culture. Colonies on PDA attaining 62 mm diam after 2 weeks, white or pale red, edge undulate, with dense aerial mycelium on the surface, fruiting bodies black to reddish brown. Conidia fusoid to ellipsoid, 4-septate, straight to slightly curved, 18.5-22 × 6-7 µm (av. = 20 × 7 µm, n = 30); basal cell obconic, hyaline, 3.5-5 µm long, with a single appendage, 2.5-6 µm; three median cells doliiform, verruculose, olivaceous with slightly red (second cell from base 4-5 µm long; third cell 4.5-5.5 µm long; fourth cell 3-5.5 µm long); apical cell conical, hyaline, 2.5-4 µm long, with 1-3 tubular appendages, 13-22.5 µm long. The morphological characters of our studied specimens fit well with Pestalotiopsis rosea (Maharachchikumbura et al. 2012). For molecular identification, the internal transcribed spacer (ITS) region, partial ß-tubulin (tub2) and translation elongation factor 1-alpha (tef1-α) genes were amplified and sequenced using primer pairs ITS5/ITS4 (White et al. 1990), T1/Bt-2b (Glass and Donaldson 1995) and EF1-728F/EF-2 (Carbone and Kohn 1999). The DNA sequences of two isolates GUCC 195257 (OP364052, OP391714, OP391713) and GUCC 195258 (OP364053, OP391716, OP391715) were deposited in GenBank. The BLAST searches revealed that these sequences had 99% (537/539 bp), 100% (453/453 bp), 99% (591/593 bp), 99% (537/539 bp), 100% (453/453 bp) and 99% (574/576 bp) nucleotide identity to the ex-type strain of P. rosea (JX399005, JX399036, JX399069), respectively. In addition, multi-locus phylogenetic analysis showed that both isolates clustered with P. rosea with full statistical support. The phylogenetic relationship of Pestalotiopsis species supported the identification of our isolates as P. rosea. In the pathogenicity test, the leaves of ten healthy 2-week-old faba bean plants were spray inoculated with a conidial suspension (1 × 106 conidia/ml) of the two isolates. Another set of five plants that were sprayed with sterilized distilled water served as the controls. Treated plants were kept at 25°C in a greenhouse with a photoperiod of 12 h and 70% relative humidity. After one week, all inoculated leaves showed symptoms similar to those of the infected faba bean observed in the field, whereas controls were symptomless. The pathogenicity test was performed twice with similar results. The fungus was reisolated from the inoculated plants and identified as P. rosea by morphological and molecular evidence, thus confirming Koch's postulates. To our knowledge, this is the first report of P. rosea causing leaf spot on faba bean in the world. Pestalotiopsis species are well-known phytopathogens that can cause a variety of diseases, including leaf spots, chlorosis, and various postharvest (Wang et al. 2019). The results of this study not only contribute to accurately identify this disease in the fields of faba bean production, but also provide an important reference for developing specific control measures.

A protoplast-based transient gene expression assay for the identification of heat and oxidative stress-regulatory genes in perennial ryegrass.

BACKGROUND: With the accumulating omics data, an efficient and time-saving transient assay to express target genes is desired. Mesophyll protoplasts, maintaining most stress-physiological responses and cellular activities as intact plants, offer an alternative transient assay to study target genes' effects on heat and oxidative stress responses. RESULTS: In this study, a perennial ryegrass (Lolium perenne L.) mesophyll protoplast-based assay was established to effectively over- or down-regulate target genes. The relative expression levels of the target genes could be quantified using RT-qPCR, and the effects of heat and H2O2-induced oxidative stress on protoplasts' viability could be quantitatively measured. The practicality of the assay was demonstrated by identifying the potential thermos-sensor genes LpTT3.1/LpTT3.2 in ryegrass that over-expressing these genes significantly altered protoplasts' viability rates after heat stress. CONCLUSION: This protoplast-based rapid stress regulatory gene identification assay was briefed as 'PRIDA' that will complement the stable genetic transformation studies to rapidly identify candidate stress-regulatory genes in perennial ryegrass and other grass species.

Associations between type 1 diabetes and pulmonary tuberculosis: a bidirectional mendelian randomization study.

BACKGROUND: Type 1 diabetes mellitus (T1DM) has been associated with higher pulmonary tuberculosis (PTB) risk in observational studies. However, the causal relationship between them remains unclear. This study aimed to assess the causal effect between T1DM and PTB using bidirectional Mendelian randomization (MR) analysis. METHODS: Single nucleotide polymorphisms (SNPs) of T1DM and PTB were extracted from the public genetic variation summary database. In addition, GWAS data were collected to explore the causal relationship between PTB and relevant clinical traits of T1DM, including glycemic traits, lipids, and obesity. The inverse variance weighting method (IVW), weighted median method, and MR‒Egger regression were used to evaluate the causal relationship. To ensure the stability of the results, sensitivity analyses assess the robustness of the results by estimating heterogeneity and pleiotropy. RESULTS: IVW showed that T1DM increased the risk of PTB (OR = 1.07, 95% CI: 1.03-1.12, P < 0.001), which was similar to the results of MR‒Egger and weighted median analyses. Moreover, we found that high-density lipoprotein cholesterol (HDL-C; OR = 1.28, 95% CI: 1.03-1.59, P = 0.026) was associated with PTB. There was no evidence of an effect of glycemic traits, remaining lipid markers, or obesity on the risk of PTB. In the reverse MR analysis, no causal relationships were detected for PTB on T1DM and its relevant clinical traits. CONCLUSION: This study supported that T1DM and HDL-C were risk factors for PTB. This implies the effective role of treating T1DM and managing HDL-C in reducing the risk of PTB, which provides an essential basis for the prevention and comanagement of concurrent T1DM and PTB in clinical practice.

A crosslinked coral-like Co@CoO/RGO nanohybrid structure with good electromagnetic wave absorption performance.

The combination of magnetic and dielectric materials followed by appropriate structure design is an effective approach to achieve high electromagnetic wave absorption properties. Here, crosslinked Co@CoO/reduced graphene oxide nanohybrids (CCRGO) were fabricated via a simple three-step method. The experimental results show that compared with previous works, the as-prepared CCRGO nanohybrids achieve higher electromagnetic wave absorption and broader effective bandwidth at a lower filler loading. The electromagnetic parameters and electromagnetic wave absorption performance could be apparently adjusted by controlling the adding content of graphene oxide (GO) and the reduction temperature. Among a series of samples, CCRGO3-650 nanohybrid yields the best electromagnetic wave absorption performance benefiting from the proper GO addition and reduction temperature. At a filler loading of 20 wt%, the maximal reflection loss reaches to -64.67 dB at a thickness of 2.53 mm and the effective bandwidth below -10 dB covers the whole X band at a thickness of 2.51 mm. The good performance may be ascribed to the advantages of the dielectric and magnetic component as well as the special crosslinked structure, which triggers a synergistic absorption mechanism including multiple reflection/scattering, interface polarization, dipole polarization, conductive loss, eddy current loss, exchange resonance in the electromagnetic wave dissipation process. The good electromagnetic wave absorption performance affirms the potential application of CCRGO nanohybrids in the field of stealth materials.

Efficacy and safety of Ophiocordyceps sinensis in the treatment of Hashimoto's thyroiditis: a systematic review and meta-analysis.

Objective: To evaluate the efficacy and safety of Ophiocordyceps sinensis (OS) preparations for the treatment of Hashimoto's thyroiditis (HT). Methods: We searched eight databases to collect randomized controlled trials (RCTs) of OS combined with a low-iodine diet or levothyroxine for HT. The search period was from inception to June 2023. Meta-analysis was performed using Revman 5.3 software after two evaluators independently screened the literature, extracted data, and evaluated the risk of bias of the included studies. The GRADE system was used to assess the certainty of evidence. Results: A total of 14 RCTs involving 1,014 patients with HT were included. Meta-analysis showed that OS preparations combined with a low-iodine diet were more effective in reducing thyroid peroxidase antibody (TPOAb) [SMD = -3.81, 95% CI (-5.07, -2.54), p < 0.00001] and thyroglobulin antibody (TgAb) [SMD = -4.73, 95% CI (-6.86, -2.61), p < 0.00001] compared to a low-iodine diet. Compared with levothyroxine treatment alone, OS preparations combined with levothyroxine further reduced TPOAb [SMD = -2.04, 95% CI (-2.82, -1.26), p < 0.00001], TgAb [SMD = -2.01, 95% CI (-2.68, -1.33), p < 0.00001], tumor necrosis factor alpha (TNF-α) [SMD = -3.40, 95% CI (-5.66, -1.14), p = 0.003], interleukin-2 (IL-2) [SMD = -2.31, 95% CI (-3.98, -0.65), p = 0.006], and interleukin-6 (IL-6) [MD = -4.16, 95% CI (-6.17, -2.15), p < 0.0001], and elevated free thyroxine (FT4) [SMD = 1.34, 95% CI (0.59, 2.08), p = 0.0004], but no significant effect on free triiodothyronine (FT3) [SMD = 0.83, 95% CI (-0.12, 1.78), p = 0.09] and thyroid stimulating hormone (TSH) [SMD = -0.80, 95% CI (-1.71, 0.11), p = 0.08]. In terms of safety, three studies reported adverse reactions in 10 patients in each of the experimental and control groups. Conclusion: OS preparations in combination with other treatments (low-iodine diet or levothyroxine) may decrease thyroid autoantibodies and inflammatory responses in patients with HT. In HT patients with hypothyroidism, the combination of the OS preparations with levothyroxine also improved FT4. However, the quality of the included studies was generally low. Moreover, the safety of OS preparations remains unclear. Therefore, more high-quality, multicenter, large-sample RCTs are needed in the future to validate the efficacy and safety of OS preparations. Systematic Review Registration: https://www.crd.york.ac.uk/prospero, identifier CRD42023432663.

Efficacy and safety of Keluoxin capsule in combination with Western medicine for diabetic kidney disease: A systematic review and meta-analysis.

Objective: Keluoxin capsule (KLXC) has been widely used in diabetic kidney disease (DKD), but its efficacy and safety have not yet been clarified. A systematic review and meta-analysis were performed to assess the efficacy and safety of KLXC for DKD. Methods: The randomized control trials (RCTs) included KLXC searched from seven major English and Chinese databases up until 3 June 2022. The methodological quality and risk of bias were assessed by version 2 of the Cochrane risk-of-bias tool (RoB 2) for the RCTs from the Cochrane Handbook. The analyses were conducted by RevMan 5.4 and Stata 17.0. Results: A total of 20 trials with 1,500 participants were identified. The meta-analysis showed that KLXC combined with Western medicine was superior to the use of Western medicine alone for DKD which included improvements in the estimated glomerular filtration rate (eGFR) [MD = 3.04, 95% CI (0.30, 5.78), p = 0.03], reduction in microalbuminuria (mALB) [MD = -25.83, 95% CI (-41.20, -10.47), p = 0.001], urinary albumin excretion rate (UAER) [SMD = -0.97, 95% CI (-1.50, -0.44), p = 0.0003], 24-h urine protein (24hUpro) [SMD = -1.31, 95% CI (-1.82, -0.80), p < 0.00001], serum creatinine (Scr) [MD = -11.39, 95% CI (-18.76, -4.02), p = 0.002], blood urea nitrogen (BUN) [MD = -1.28, 95% CI (-1.67, -0.88), p < 0.00001], fasting blood glucose (FBG) [MD = -0.51, 95% CI (-0.90, -0.11), p = 0.01], total cholesterol (TC) [MD = -1.04, 95% CI (-1.40, -0.68), p < 0.00001], triglycerides (TG) [MD = -0.36, 95% CI (-0.50, -0.23), p < 0.00001], and low-density lipoprotein cholesterol (LDL) [MD = -0.39, 95% CI (-0.71, -0.07), p = 0.02]. Results showed no statistically significant difference in glycated hemoglobin (HbA1c) (p = 0.14) or adverse events (p = 0.81) between the two groups. Conclusion: The combination of KLXC and Western medicine had a positive effect on DKD. However, due to the high clinical heterogeneity and low quality of included studies, further standardized, large-scale, rigorously designed RCTs for DKD in the definitive stage are still necessary to achieve more accurate results. Systematic Review Registration: https://inplasy.com/inplasy-2021-11-0067/, identifier INPLASY 2021110067.

A bibliometric analysis of RNA methylation in diabetes mellitus and its complications from 2002 to 2022.

Background: RNA methylation has emerged as an active research field in diabetes mellitus (DM) and its complications, while few bibliometric analyses have been performed. We aimed to visualize the hotspots and trends using bibliometric analysis to provide a comprehensive and objective overview of the current search state in this field. Methods: The articles and reviews regarding RNA methylation in DM and its complications were from the Web of Science Core Collection. A retrospective bibliometric analysis and science mapping was performed using the CiteSpace software to plot the knowledge maps and predict the hotspots and trends. Results: Three hundred seventy-five qualified records were retrieved. The annual publications gradually increased over the past 20 years. These publications mainly came from 66 countries led by Canada and 423 institutions. Leiter and Sievenpiper were the most productive authors, and Jenkins ranked first in the cited authors. Diabetes Care was the most co-cited journal. The most common keywords were "Type 2 diabetes", "cardiovascular disease", "diabetes mellitus", and "n 6 methyladenosine". The extracted keywords mainly clustered in "beta-cell function", "type 2 diabetes", "diabetic nephropathy", "aging", and "n6-methyladenosine". N6-methyladenosine (m6A) in DM and its complications were the developing areas of study. Conclusion: Studies on RNA methylation, especially m6A modification, are the current hotspots and the future trends in type 2 diabetes (T2D) and diabetic nephropathy (DN), as well as a frontier field for other complications of DM. Strengthening future cooperation and exchange between countries and institutions is strongly advisable to promote research developments in this field.

Multigene Phylogeny, Diversity and Antimicrobial Potential of Endophytic Sordariomycetes From Rosa roxburghii.

Rosa roxburghii Tratt. is widely applied in food, cosmetics, and traditional medicine, and has been demonstrated to possess diverse bioactivities. Plant endophytic fungi are important microbial resources with great potential for application in many fields. They not only establish mutualistic symbiosis with host plants but also produce a variety of bioactive compounds. Therefore, in the present study, endophytic fungi were isolated from R. roxburghii, the diversity and antimicrobial activities were evaluated. As a result, 242 strains of endophytic Sordariomycetes were successfully isolated. Multigene phylogenetic analyses showed that these isolates included eight orders, 19 families, 33 genera. The dominant genera were Diaporthe (31.4%), Fusarium (14.4%), Chaetomium (7.9%), Dactylonectria (7.0%), Graphium (4.5%), Colletotrichum (4.1%), and Clonostachys (4.1%). For different tissues of R. roxburghii, alpha diversity analysis revealed that the diversity of fungal communities decreased in the order of root, fruit, stem, flower, leaf, and seed, and Clonostachys and Dactylonectria exhibited obvious tissue specificity. Meanwhile, functional annotation of 33 genera indicated that some fungi have multitrophic lifestyles combining endophytic, pathogenic, and saprophytic behavior. Additionally, antimicrobial activities of endophytic Sordariomycetes against Lasiodiplodia theobromae, Botryosphaeria dothidea, Colletotrichum capsici, Pyricularia oryzae, Rhizoctonia solani, Fusarium oxysporum, Pseudomonas syringae, Pantoea agglomerans, Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa were screened. Dual culture test assays showed that there were 40 different endophytic species with strong inhibition of at least one or moderate inhibition of two or more against the 12 tested strains. The results from the filter paper diffusion method suggested that extracellular metabolites may be more advantageous than intracellular metabolites in the development of antimicrobial agents. Eleven isolates with good activities were screened. In particular, Hypomontagnella monticulosa HGUP194009 and Nigrospora sphaerica HGUP191020 have shown promise in both broad-spectrum and intensity. Finally, some fungi that commonly cause disease have been observed to have beneficial biological activities as endophytic fungi. In conclusion, this study showed the species composition, alpha diversity, and lifestyle diversity of endophytic Sordariomycetes from R. roxburghii and demonstrated these isolates are potential sources for exploring antimicrobial agents.

Ascodesmisrosicola sp. nov. and Talaromycesrosarhiza sp. nov., two endophytes from Rosaroxburghii in China.

BACKGROUND: Rosaroxburghii Tratt., a deciduous shrub of the family Rosaceae, is usually used as food and medicinal materials and also cultivated as an ornamental. Plant endophytic fungi are a large class of microbial resources not fully researched, with great potential applications. Two strains of Ascodesmis and Talaromyces were isolated during a survey of biodiversity on endophytic fungi of R.roxburghii in China. Multigene phylogenetic analyses showed that each of the two fungi formed a distinct lineage and separated from known congeneric species and they are proposed as two novel taxa. NEW INFORMATION: Ascodesmisrosicola sp. nov. usually has one or two conspicuous simple or branched ridges extending to the majority of the ascospore surface and remarkably small asci, distinguishing it from the previously-described species in the genus Ascodesmis. Talaromycesrosarhiza sp. nov., of the section Talaromyces, is closely related to T.francoae. It differs from the latter by having both monoverticillate and biverticillate conidiophores, while those of T.francoae are biverticillate. Both novel endophytes are illustrated and described.