First Report of Anthracnose Leaf Spot Caused by Colletotrichum americae-borealis on Potentilla anserina in China.

Potentilla anserina L. has an abundance of bioactive compounds and is widely recognized for its diverse applications in traditional medicine and as a food. In August 2023, typical symptoms of anthracnose were observed in 80% of P. anserina plants in Harbin, China. Symptoms, characterized by reddish-brown spots, tend to occur more frequently on leaves closer to the ground. They initially appeared as oval or irregular circles, measuring 1 to 3 mm in diameter, and later merged into larger patches surrounded by chlorotic areas on the leaves. Twenty leaves exhibiting characteristic symptoms were sampled. Each leaf was sectioned into 5×5 mm pieces at the interface between the diseased and healthy tissues. The sections were disinfected sequentially with 75% ethanol for 30 s, followed by 1% NaClO for 2 min, rinsed three times in sterilized distilled water. Post air-drying, samples were cultured on potato dextrose agar (PDA) plates and incubated at 26°C in the dark for 5 d, yielding nine morphologically similar single-spore isolates (JTC1 to JTC9). The colonies initially displayed gray aerial mycelia, becoming pale brown, accompanied by numerous black microsclerotia. The acervuli appeared black, protruded from the surface of the medium, and were adorned with dark brown setae. Setae (n=50) ranged from 58.4 to 188.2 µm in length, appearing dark brown to black, with smooth walls, rounded tips, swollen bases, and containing 1 to 4 septa. The conidia were hyaline, aseptate, cylindrical to spindle-shaped, with blunt and rounded ends, measuring 13.7 to 18.3 µm in length and 3.4 to 4.3 µm in width (n=50). Morphological analysis indicated a close affinity with Colletotrichum americae-borealis (Damm et al. 2014). For molecular identification, genomic DNA was extracted from three representative isolates (JTC1, JTC2, and JTC3).The ITS, HIS3，GAPDH, and ACT genes were amplified and sequenced using the primers described previously by Damm et al. (2014). The sequences were submitted to GenBank (ITS: PP338190 to PP338192; HIS3: PP355770 to PP355772; GAPDH: PP355773 to PP355775; ACT: PP355776 to PP355778). BLAST analysis showed 99 to 100% identity with C. americae-borealis type strain CBS 136232 (GenBank accessions: KM105224, KM105364, KM105579, and, KM105434, respectively). Multigene phylogenetic analysis positioned the three isolates close to C. americae-borealis. Pathogenicity tests were performed twice on 6-week-old P. anserina seedlings (cv. Qinghai Juema 1) in a greenhouse. A conidial suspension of the JTC1 isolate (1×105 conidia/ml) was sprayed applied to ten pots, each containing two seedlings, and the plants in the control pots were sprayed with sterile distilled water. Two weeks after inoculation under greenhouse conditions (26/22°C day/night temperature, 12-hour photoperiod, 90% relative humidity), the inoculated seedlings exhibited brown spots and necrotic lesions similar to those observed in the field, C. americae-borealis was successfully reisolated from these symptomatic tissues. To the best of our knowledge, this is the first report of C. americae-borealis causing leaf spot on P. anserina in China. Anthracnose caused by C. americae-borealis is associated with leaf spot disease in oats (Wang et al. 2022), alfalfa (Li et al. 2021), and licorice (Lyu et al.2020). However, C. americae-borealis poses a significant threat to P. anserina in China as well, highlighting the urgent need to develop effective disease management strategies.

Genome-wide identification and characterization of NAC transcription factor family members in Trifolium pratense and expression analysis under lead stress.

BACKGROUND: The NAC TF family is widely involved in plant responses to various types of stress. Red clover (Trifolium pratense) is a high-quality legume, and the study of NAC genes in red clover has not been comprehensive. The aim of this study was to analyze the NAC gene family of red clover at the whole-genome level and explore its potential role in the Pb stress response. RESULTS: In this study, 72 TpNAC genes were identified from red clover; collinearity analysis showed that there were 5 pairs of large fragment replicators of TpNAC genes, and red clover was found to be closely related to Medicago truncatula. Interestingly, the TpNAC genes have more homologs in Arabidopsis thaliana than in soybean (Glycine max). There are many elements in the TpNAC genes promoters that respond to stress. Gene expression analysis showed that all the TpNAC genes responded to Pb stress. qRT-PCR showed that the expression levels of TpNAC29 and TpNAC42 were significantly decreased after Pb stress. Protein interaction network analysis showed that 21 TpNACs and 23 other genes participated in the interaction. In addition, the TpNAC proteins had three possible 3D structures, and the secondary structure of these proteins were mainly of other types. These results indicated that most TpNAC members were involved in the regulation of Pb stress in red clover. CONCLUSION: These results suggest that most TpNAC members are involved in the regulation of Pb stress in red clover. TpNAC members play an important role in the response of red clover to Pb stress.

Identification and characterization of Colletotrichum truncatum and C. destructivum causing stem rot of white clover in China.

White clover (Trifolium repens L.) is an excellent quality forage legume species with superior planting efficiency, which reduces the cost of artificial weeding and nitrogen fertilizer inputs and has feeding and economic value. However, from June to September 2022, severe stem rot affected the yield and quality of white clover crops in Harbin, Heilongjiang Province, China. The aim of this study was to identify the causative agents of the disease. Overall, Colletotrichum truncatum (6 isolates) and C. destructivum (10 isolates) were obtained from rotten white clover stems and identified according to morpho-molecular characteristics and phylogenetic analyses. Pathogenicity tests of the isolates revealed that C. destructivum had a higher pathogenicity to white clover than C. truncatum. In addition, all isolates were highly pathogenic to broad bean, fodder soybean, soybean, pak choi, and chickpea, pathogenic to mint, and did not infect corn, wheat, or cilantro. C. destructivum and C. truncatum isolates were very sensitive to tebuconazole and pyraclostrobin, with EC50 values of 0.54 to 0.70 µg/ml and 0.42 to 0.62 µg/ml, respectively, efficacies ranging between 93.2 to 94.9% and 90.2 to 95.2% at 600 µg/ml and 450 µg/ml, respectively, and EC90 values of 1.88 to 13.36 µg/ml and 1.32 to 23.39 µg/ml, respectively. Therefore, intercropping of host and non-host plants and chemicals can be considered to control stem rot in white clover. These results provide a basis for controlling C. destructivum and C. truncatum in white clover in China.

Identification and Evolutionary Analysis of the Auxin Response Factor (ARF) Family Based on Transcriptome Data from Caucasian Clover and Analysis of Expression Responses to Hormones.

Caucasian clover (Trifolium ambiguum M. Bieb.) is an excellent perennial plant in the legume family Fabaceae, with a well-developed rhizome and strong clonal growth. Auxin is one of the most important phytohormones in plants and plays an important role in plant growth and development. Auxin response factor (ARF) can regulate the expression of auxin-responsive genes, thus participating in multiple pathways of auxin transduction signaling in a synergistic manner. No genomic database has been established for Caucasian clover. In this study, 71 TaARF genes were identified through a transcriptomic database of Caucasian clover rhizome development. Phylogenetic analysis grouped the TaARFs into six (1-6) clades. Thirty TaARFs contained a complete ARF structure, including three relatively conserved regions. Physical and chemical property analysis revealed that TaARFs are unstable and hydrophilic proteins. We also analyzed the expression pattern of TaARFs in different tissues (taproot, horizontal rhizome, swelling of taproot, rhizome bud and rhizome bud tip). Quantitative real-time RT-PCR revealed that all TaARFs were responsive to phytohormones (indole-3-acetic acid, gibberellic acid, abscisic acid and methyl jasmonate) in roots, stems and leaves. These results helped elucidate the role of ARFs in responses to different hormone treatments in Caucasian clover.

Construction of Porous Carbon Nanosheet/Cu2S Composites with Enhanced Potassium Storage.

Porous C nanosheet/Cu2S composites were prepared using a simple self-template method and vulcanization process. The Cu2S nanoparticles with an average diameter of 140 nm are uniformly distributed on porous carbon nanosheets. When used as the anode of a potassium-ion battery, porous C nanosheet/Cu2S composites exhibit good rate performance and cycle performance (363 mAh g-1 at 0.1 A g-1 after 100 cycles; 120 mAh g-1 at 5 A g-1 after 1000 cycles). The excellent electrochemical performance of porous C nanosheet/Cu2S composites can be ascribed to their unique structure, which can restrain the volume change of Cu2S during the charge/discharge processes, increase the contact area between the electrode and the electrolyte, and improve the electron/ionic conductivity of the electrode material.

Effects of different concentrations of biochar amendments and Pb toxicity on rhizosphere soil characteristics and bacterial community of red clover (Trifolium pretense L.).

Amending soil with biochar can reduce the toxic effects of heavy metals (HM) on plants and the soil. However, the effects of different concentrations of biochar on the properties and microbial activities in lead (Pb)-contaminated soils are unclear. In this study, two Pb concentrations were set (low, 1000 mg/kg; high, 5000 mg/kg), and five corn straw biochar (CSB) concentrations (0, 2.5, 5, 10 and 15%) were used to determine the response of the growth and rhizosphere of red clover (Trifolium pretense L.) (in terms of soil properties and bacteria) to CSB and Pb application. The results showed that 5% CSB better alleviated the toxicity of Pb on the shoot length of red clover, the biomass increased by 74.55 and 197.76% respectively and reduced the enrichment factor (BCF) and transport factor (TF) of red clover. Pb toxicity reduced soil nutrients, catalase (CAT), acid phosphatase (ACP) and urease activity, while the addition of CSB increased soil pH, soil organic matter (SOM) content and soil enzyme activity. 16S rDNA amplicon sequencing analysis showed that Pb toxicity reduced the diversity of rhizosphere bacteria in red clover and reduced the relative abundance of plant growth-promoting rhizobacteria such as Gemmatimonas, Devosia and Bryobacter. Spearman correlation analysis showed that the addition of alkaline CSB restored the relative abundance of rhizobacteria positively correlated with pH, such as Chitinophaga, Sphingomonas, Devosia and Pseudomonas, and thus restored the rhizosphere soil environment. This study demonstrates that 5% CSB can better alleviate the toxicity of Pb to red clover and soil. We also provide a theoretical basis for the subsequent use of beneficial bacteria to regulate the repair efficiency of red clover.

Trumpet-Like ZnS@C Composite for High-Performance Potassium Ion Battery Anode.

ZnS has acquired increasing attention for high-performance PIBs anode because of its remarkable theoretical capacity, and redox reversibility for conversion reaction. However, the larger volume variation and delayed reaction kinetics for the ZnS in the discharge/charge processes lead to pulverization and severe capacity degradation. Herein, the trumpet-like ZnS@C composite was synthesized by template method by using sodium citrate as carbon source followed by vulcanization process. As potassium ion batteries (PIB) anode, ZnS@C composite exhibits good rate performance and long life (stable reversible capacity of 107.8 mAh/g over 2000 charge-discharge cycles at 5 A/g and high reversible capacity of 310 mAh/g at 0.1 A/g). The outstanding electrochemical performance of the ZnS@C composite is ascribed to its unique structure, which can mitigate the volume expansion of ZnS in the charge discharge process, expand the contact area between the electrode and electrolyte, and improve the conductivity of electrode materials by the introduction of carbon layer. This method of synthesizing trumpet-like ZnS@C composite provides an important strategy for obtaining potassium ion batteries anode with long cycle.

Fabrication of Nb2 O5 /Carbon Submicrostructures for Advanced Lithium-Ion Battery Anodes.

Nb2 O5 possesses superior fast Li+ storage capability for LIB anodes, benefiting from its fast pseudocapacitive behavior and low volumetric change within the cycling processes. However, the poor electric conductivity for Nb2 O5 restricts its reaction kinetics and rate property. Herein, Nb2 O5 /carbon (C) submicrostructures are fabricated by solvothermal method followed by calcination process. The Nb2 O5 /C submicrostructures exhibit outstanding rate behavior and cyclic performance (332 (194) mAh g-1 after 1000 cycles at 1 (5) A g-1 ). The superior electrochemical property is attributed to the distinctive structure for Nb2 O5 /C submicrostructures, in which Nb2 O5 nanoparticles uniformly distributed within Nb2 O5 /C composite can protect Nb2 O5 nanoparticles from agglomeration, and the porous carbon matrix can enhance electron/ion conductivity. This work furnishes a novel strategy for fabricating Nb2 O5 /C submicrostructures with superior Li+ storage performance, which can be potentially used to design other metal oxide/C submicrostructures for second battery anode.

Construction of Fe7Se8@Carbon nanotubes with enhanced sodium/potassium storage.

The Fe7Se8@Carbon (C) nanotubes are successfully synthesized using Fe3O4@C nanotubes as sacrificial templates. Fe7Se8@C nanotubes exhibit excellent rate behaviour and maintainable capacity (319 mAh g-1 at 2 A g-1 upon 720 cycles), when utilized as SIBs anode. Moreover, for PIBs anode, Fe7Se8@C nanotubes also exhibit outstanding rate behaviour and maintainable capacity 222 mAh g-1 at 2 A g-1 upon 500 cycles). The superior electrochemical performance of Fe7Se8@C nanotubes is ascribed to the unique structure, where the hierarchical hollow tubular characteristic of Fe7Se8@C composites can mitigate the volume expansion of Fe7Se8 and supply effective transmission paths for both Na+(K+) and electrons within repeated cycle processes, and additionally, N-doped carbon layer can further protect the integrality of Fe7Se8@C nanotubes from destruction within the cycle processes, and enhance electronic conductivity.