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This research aimed to investigate the impacts of a combination of rice husk biochar and organic fertilizer on the physical and chemical properties of soil, the population of soil bacteria, the relative chlorophyll content of leaves, the development of soybean root nodules, and yield components under strongly acid soil conditions. A greenhouse and pot experiment was designed using a randomize complete block design with factorial 2 × 3 treatments and three replications. The experimental treatments comprised two rates of biochar (35 and 70 g/pot) and three rates of organic fertilizer (70, 105, and 140 g/pot). After 100 days of amendment of strongly acidic soils, the results showed that application of treatments B35F70 and B70F140 increased soil pH by 16.80% compared to the control group (CK). On the other hand, treatments B35F140 and B70F105 resulted in an increase of soil electrical conductivity by 66.67% compared to CK. In addition, after 100 days of amendment with treatments B35F105, B35F105, B35F140, B70F105, B70F70, B70F70, and B35F140, organic matter, available phosphorous (P), potassium (K), calcium (Ca), magnesium (Mg), copper (Cu), and zinc (Zn), organic matter, available phosphorous (P), potassium (K), calcium (Ca), magnesium (Mg), copper (Cu), and zinc (Zn), significantly increased when compared to the control group (CK). Treatment B35F140 increased relative leaf chlorophyll content and soybean seed weight per plant by 60.76% and 100.56%, respectively when compared to the CK. Furthermore, treatment B35F70 produced 125% more root nodules than CK. Moreover, each amended strongly acid soil resulted with a significant upsurge in total soil bacteria compared to the CK. Overall, statistics proved that a combination of biochar and organic fertilizer improved soil properties and soybean agronomic attributes. [ABSTRACT FROM AUTHOR]

Efficient and sustainable food production is crucial in global agricultural development. Overuse of chemical fertilizers leads to soil acidification, destruction of soil properties, and harm to soil micro-organisms. Plant growth-promoting rhizobacteria (PGPR) have emerged as a solution, enhancing soil fertility and crop yields while reducing chemical fertilizer dependency and disease occurrence. In this study, Bacillus strains KHC2, KHH5, and KHH13, isolated from organic rice field soils in Taiwan, were identified through molecular techniques as B. velezensis (KHC2, KHH13) and B. amyloliquefaciens (KHH5). The strains exhibited various hydrolytic enzymes (including protease, cellulase, amylase, and lecithinase), with KHH13 showing the highest phosphate solubilization (2186.1 µg mL−1 day−1) and indole-3-acetic acid (IAA) production (63.067 ± 0.595 ppm mL−1). These properties indicate KHH13's potential as a bio-enhancer for plant growth. Therefore, we hypothesized that KHH13 can enhance plant growth and control soil-borne diseases. A greenhouse experiment demonstrated that KHH13, KHC2, and KHH5 effectively promoted the growth of red lettuce, with KHH13 showing superior efficacy. The study also found KHH13's treatment enhanced the growth of various vegetables, including tomato, cucumber, and red lettuce. In terms of disease control, KHH13 significantly reduced Fusarium wilt in cucumbers, as evidenced by the reduction in disease index from 74.33% to 41.67% after KHH13 treatment. The treatment group displayed better plant growth, including plant height and fresh weight, compared to the control group in the greenhouse experiment. Furthermore, oral and pulmonary acute toxicity analysis in rats showed no adverse effects on rat weight or mortality, indicating KHH13's safety for mammalian use. These findings suggest B. velezensis KHH13 as a safe, effective, and sustainable biological agent for enhancing vegetable growth and controlling soil-borne diseases, with potential applications in sustainable agriculture. [ABSTRACT FROM AUTHOR]

Melon (Cucumis melo L.) is a global commercial crop that is sensitive to seed-borne wilt infections caused by Fusarium oxysporum f. sp. melonis (Fom). To address the challenge of detecting Fom contamination, we designed a probe-based real-time PCR method, TDCP2, in combination with rapid or column-based DNA extraction protocols to develop reliable molecular detection methods. Utilizing TDCP2, the detection rate reached 100% for both artificially Fom-inoculated (0.25–25%) and pod-inoculated melon seeds in conjunction with DNA samples from either the rapid or column-based extraction protocol. We performed analyses of precision, recall, and F1 scores, achieving a maximum F1 score of 1 with TDCP2, which highlights the robustness of the method. Additionally, intraday and interday assays were performed, which revealed the high reproducibility and stability of column-based DNA extraction protocols combined with TDCP2. These metrics confirm the reliability of our developed protocols, setting a foundation for future enhancements in seed pathology diagnostics and potentially broadening their applicability across various Fom infection levels. In the future, we hope that these methods will reduce food loss by improving the control and management of melon diseases. [ABSTRACT FROM AUTHOR]

We developed an insulated isothermal PCR (iiPCR) method for the efficient and rapid detection of Fusarium oxysporum (Fo), which is a fungus that infects various hosts and causes severe crop losses. The Fo iiPCR method was sensitive enough to detect up to 100 copies of standard DNA template and 10 fg of Fo genomic DNA. In addition, it could directly detect 1 pg of mycelium and 10 spores of Fo without DNA extraction. Our study compared the performance of Fo iiPCR to that of three published in planta molecular detection methods—conventional PCR, SYBR green-based real-time PCR, and hydrolysis probe-based real-time PCR—in field detection of Fo. All diseased field samples yielded positive detection results with high reproducibility when subjected to an Fo iiPCR test combined with a rapid DNA extraction protocol compared to Fo iiPCR with an automated magnetic bead-based DNA extraction protocol. Intraday and interday assays were performed to ensure the stability of this new rapid detection method. The results of detection of Fo in diseased banana pseudostem samples demonstrated that this new rapid detection method was suitable for field diagnosis of Fusarium wilt and had high F1 scores for detection (the harmonic mean of precision and recall of detection) for all asymptomatic and symptomatic Fo-infected banana samples. In addition, banana samples at four growth stages (seedling, vegetative, flowering and fruiting, and harvesting) with mild symptoms also showed positive detection results. These results indicate that this new rapid detection method is a potentially efficient procedure for on-site detection of Fo. [ABSTRACT FROM AUTHOR]

A new fault detection/location technique with consideration of arcing fault discrimination based on phasor measurement units for extremely high voltage/ultra-high voltage transmission lines is presented in this two-paper set. Part I of this two-paper set is mainly aimed at theory and algorithm derivation. The proposed fault detection technique for both arcing and permanent faults is achieved by a combination of a fault detection index |M| and a fault location index |D|, which are obtained by processing synchronized fundamental phasors. One is to detect the occurrence of a fault and the other is to distinguish between in-zone and out-of-zone faults. Furthermore, for discriminating between arcing and permanent faults, the proposed technique estimates the amplitude of arc voltage by least error squares method through the measured synchronized harmonic phasors caused by the nonlinear are behavior. Then, the discrimination will be achieved by comparing the estimated amplitude of are voltage to a given threshold value. In addition, in order to eliminate the error caused by exponentially decaying dc offset on the computations of fundamental and harmonic phasors, an extended discrete Fourier transform algorithm is also presented. Index Terms--Arcing fault discrimination, extended discrete Fourier transform (EDFT), fault detection, fault location, phasor measurement unit (PMU).

Rhizoctonia solani (Rs), a soil-borne fungal pathogen, can result in rice sheath blight (ShB), which causes yield loss. To prevent outbreaks of ShB and enhance the sustainability of rice production, it is critical to develop a rapid ShB detection method for specific, fast, and on-site disease management. In this study, a reagent for the rapid extraction of this pathogen was developed for on-site detection. The specificity and sensitivity of a novel SMS RS1-F/SMS RS1-R primer set and a ITS1/GMRS-3 reference primer set were tested, while four different extraction protocols for ShB were developed. Moreover, intraday and interday assays were performed to evaluate the reproducibility of the detection methods developed. The results indicated that all of the developed protocols are suitable for use in detecting ShB. In addition, all the samples of infected rice yielded positive Rs detection results when subjected to TaqMan probe-based real-time PCR and SYBR green-based real-time PCR (SMS RS1-F/SMS RS1-R) tests in which automatic magnetic bead-based DNA extraction was performed. These results indicated that the two molecular detection protocols were suitable for the field diagnosis of ShB for all asymptomatic and symptomatic rice samples. [ABSTRACT FROM AUTHOR]

Silver particles were prepared by dewetting Ag films coated on glass using a fiber laser. The size of the particles was controlled in the range of 92 nm–1.2 μm by adjusting the thickness of the Ag film. The structural properties and surface roughness of the particles were evaluated by means of scanning electron microscopy. In addition, the antifungal activity of the Ag particles was examined using spore suspensions of Colletotrichumgloeosporioides. It is shown that particles with a size of 1.2 μm achieved 100% inhibition of conidia growth of C.gloeosporioides after a contact time of just 5 min. Furthermore, the smaller particles also achieved good antifungal activity given a longer contact time. Similar results were observed for spore germination and pathogenicity tests performed on mango fruit and leaves. Overall, the results confirm that Ag particles have an excellent antifungal effect on C. gloeosporioides. [ABSTRACT FROM AUTHOR]

Banana (Musa sp.) is cultivated worldwide and is one of the most popular fruits. The soil-borne fungal disease Fusarium wilt of banana (FWB), commonly known as Panama disease, is caused by Fusarium oxysporum f. sp. cubense (Foc) and is a highly lethal vascular fungal disease in banana plants. Raman spectroscopy, an emerging laser-based technology based on Raman scattering, has been used for the qualitative characterization of biological tissues such as foodborne pathogens, cancer cells, and melamine. In this study, we describe a Raman spectroscopic technique that could potentially be used as a method for diagnosing FWB. To that end, the Raman fingerprints of Foc (including mycelia and conidia) and Foc-infected banana pseudostems with varying levels of symptoms were determined. Our results showed that eight, eleven, and eleven characteristic surface-enhanced Raman spectroscopy peaks were observed in the mycelia, microconidia, and macroconidia of Foc, respectively. In addition, we constructed the Raman spectroscopic fingerprints of banana pseudostem samples with varying levels of symptoms in order to be able to differentiate Foc-infected bananas from healthy bananas. The rate at which FWB was detected in asymptomatic Foc-infected samples by using the spectral method was 76.2%, which was comparable to the rates previously reported for other FWB detection methods based on real-time PCR assays, suggesting that the spectral method described herein could potentially serve as an alternative tool for detecting FWB in fields. As such, we hope that the developed spectral method will open up new possibilities for the on-site diagnosis of FWB. [ABSTRACT FROM AUTHOR]

Part I of this paper set sets forth theory and algorithms for adaptive fault detection/location technique, which is based on Phasor Measurement Unit (PMU). This paper is Part II of this paper set. A new timing device named 'Global Synchronism Clock Generator, GSCG' including its hardware and software design is described in this paper. Experimental results show that the synchronized error of rising edge between the two GSCGs clock is well within 1 ps when the clock frequency is below 2.499 MHz. The measurement results between Chung-Jeng and Chang-Te 161 kV substations of Taiwan Power company by PMU equipped with GSCG is presented and the accuracy for estimating parameters of line is verified. The new developed DFT based method (termed as Smart Discrete Fourier Transform, SDFT) and line parameter estimation algorithm are combined with PMU configuration to form the adaptive fault detector/locator system. Simulation results have shown that SDFT method can extract exact phasors in the presence of frequency deviation and harmonics. The parameter estimation algorithm can also trace exact parameters very well. The SDFT method and parameter estimation algorithm can achieve accuracies of up to 99.999% and 99.99%, respectively. The EMTP is used to simulate a 345 kV transmission line of Taipower System. Results have shown that the proposed technique yields correct results independent of fault types and is insensitive to the variation of source impedance, fault impedance, and line loading. The accuracy of fault location estimation achieved can be up to 99.9% for many simulated cases. The proposed technique will be very suitable for implementation in an integrated digital protection and control system for transmission substations. Index Terms--Discrete Fourier transforms (DFT), fault detection/location index, phasor measurement unit (PMU).

An adaptive fault detection/location technique based on Phasor Measurement Unit (PMU) for an EHV/UHV transmission line is presented in this two paper set. This paper is Part I of this set. A fault detection/location index in terms of Clarke components of the synchronized voltage and current phasors is derived. The line parameter estimation algorithm is also developed to solve the uncertainty of parameters caused by aging of transmission lines. This paper also proposes a new Discrete Fourier Transform (DFT) based algorithm (termed as Smart Discrete Fourier Transform, SDFT) to eliminate system noise and measurement errors such that extremely accurate fundamental frequency components can be extracted for calculation of fault detection/location index. The EMTP was used to simulate a high voltage transmission line with faults at various locations. To simulate errors involved in measurements, Gaussian-type noise has been added to the raw output data generated by EMTP. Results have shown that the new DFT based method can extract exact phasors in the presence of frequency deviation and harmonics. The parameter estimation algorithm can also trace exact parameters very well. The accuracy of both new DFT based method and parameter estimation algorithm can achieve even up to 99.999% and 99.99% respectively, and will be presented in Part H. The accuracy of fault location estimation by the proposed technique can achieve even up to 99.9 % in the performance evaluation, which is also presented in Part II. Index Terms--Fault Detection/Location Index, Discrete Fourier Transforms (DFT), Phasor Measurement Unit (PMU).

In this paper, atmospheric-pressure air and oxygen corona plasma jets under water surface have been used to generate plasma-activated water (PAW) to inactivate colletotrichum gloeosporioides (C. gloeosporioides) suspension. The atmospheric-pressure plasma jets in the water were characterized by measuring: 1) the electrical and optical properties for evaluating the plasma efficiency and 2) the physicochemical properties of PAW, including pH value, oxidation-reduction potential value, and the concentration of hydrogen peroxide, nitrite, and ozone. The aim of this paper is to investigate the effects of long-lived reactive oxygen and nitrogen species (RONS) and environmental factor (acidity) in PAW on fungicidal property. The results of antifungal activity show that the inactivation rate of C. gloeosporioides is higher using air-PAW than oxygen-PAW, and increases with increasing reaction time when preparation time of PAW is fixed as 30 min. This correlates well with measurements of the aqueous-phase long-lived RONS concentration such as nitrate and ozone, suggesting these species play crucial roles in the inactivation process. [ABSTRACT FROM AUTHOR]

This study developed a novel and inexpensive detection method based on a TaqMan probe-based insulated isothermal polymerase chain reaction (iiPCR) method for the rapid detection of Panama disease caused by Fusarium oxysporum f. sp. cubense (Foc) race 4, which is currently among the most serious fungal vascular diseases worldwide. By using the portable POCKIT™ device with the novel primer set iiFoc-1/iiFoc-2, the Foc race 4 iiPCR assay (including DNA amplification and signal monitoring) could be completed within one hour. The developed Foc race 4 iiPCR assay is thus a user-friendly and efficient platform designed specifically for the detection of Foc race 4. The detection limit of this optimized Foc iiPCR system was estimated to be 1 copy of the target standard DNA as well as 1 fg of the Foc genomic DNA. This approach can serve as a rapid detection method for in planta detection of Foc race 4 in field-infected banana. It was concluded that this molecular detection procedure based on iiPCR has good potential for use as an efficient detection method. [ABSTRACT FROM AUTHOR]

All plants synthesize multiple families of small heat shock proteins (sHSP). A cDNA clone, Oshsp18.0-CII, encoding an 18.0 kDa class II sHSP was isolated from rice previously. The function of the rice class II sHSP was studied by overproduction of the Oshsp18.0-CII fusion protein in transformed Escherichia coli cells. The results suggest that heterologous expression of the Oshsp18.0-CII fusion protein increases thermotolerance of E. coli cells in vivo and provided thermoprotection to E. coli soluble proteins in vitro. The survival rate of Oshsp 18.0-CII fusion protein-accumulating cells treated at 50°C for 1 h was almost 1000- fold higher than that of the control cells transformed with pET32a expression vector. Overproduction of the Oshsp18.0-CII fusion protein in E. coli also confers tolerance of E. coli cells to ultraviolet (UV) irradiation. The post-UV survival of the Oshsp18.0-CII fusion protein-accumulating cells is about 7.29-fold over that of the control cells transformed with pET32a expression vector when exposed to 1700 μJ of UV. There is almost no post-UV survival (≦0.4%) in the untransformed cells after exposing to 900 μJ UV light. [ABSTRACT FROM AUTHOR]

The theory and algorithms of the proposed technique have been presented in Part I of this two-paper set. In Part II of this two-paper set, the proposed technique is evaluated by considerable simulation cases simulated by the Mattab/Power system Blockset simulator. For the proposed fault detector, the trip time achieved can be up to 3.25 ms and the average value of trip times is about 8 ms for both permanent and arcing faults on transmission lines. For the proposed fault locator, the accuracy can be up to 99.99% and the error does not exceed 0.45%. Moreover, the proposed arcing fault discriminator can discriminate between arcing and permanent faults within four cycles after fault inception. It has proven to be an effective tool to block reclosing on the permanent faults in the computer simulations. The simulation results also demonstrate that the presented extended discrete Fourier transform algorithm eliminates effectively the error caused by exponentially decaying dc offset on fundamental and harmonic phasor computations. Finally, a test case using the real-life measured data proves the feasibility of the proposed technique. [ABSTRACT FROM AUTHOR]

Red crown rot (RCR), caused by the soilborne fungus Calonectria ilicicola , is an emerging soybean disease in Taiwan, and fungicide screening is desired to identify effective management for C. ilicicola . This study screened 11 fungicides, including azoxystrobin, boscalid, cyprodinil, cyprodinil + fludioxonil, difenoconazole, fluopyram, flutolanil, mancozeb, prochloraz, pyraclostrobin, and tebuconazole, for their inhibitory effects on the mycelial growth of 10 C. ilicicola field isolates. Subsequently, a microplate-based high-throughput screening (MHTS) method was established to measure the fungicide sensitivity in a population composed of 80 C. ilicicola isolates to three effective fungicides, cyprodinil + fludioxonil, fluopyram, and tebuconazole. The MHTS was optimized for multiple factors, including the optical scanning pattern, absorption wavelength, conidial concentration, and measurement timing based on the quality controls of Z' factor and the log-phase growth curve. The population mean EC 50 estimated by MHTS were 0.14, 2.34, and 2.46 ppm to cyprodinil + fludioxonil, fluopyram, and tebuconazole, respectively. In addition to the in vitro assessment, fungicide efficacy was evaluated by coating cyprodinil + fludioxonil, fluopyram, or tebuconazole on soybean seeds in the pot assay. The results showed that cyprodinil + fludioxonil significantly reduced both postemergence damping-off and disease severity, while fluopyram and tebuconazole reduced only the postemergence damping-off but not disease severity. Based on the MHTS and the pot assay results, this study demonstrated cyprodinil + fludioxonil to be a potential fungicide to manage soybean RCR., Competing Interests: The author(s) declare no conflict of interest.

Brown blight, a destructive foliar disease of tea, has become a highly limiting factor for tea cultivation in Taiwan. To understand the population composition of the causal agents ( Colletotrichum spp.), the fungal diversity in the main tea-growing regions all over Taiwan was surveyed from 2017 to 2019. A collection of 139 Colletotrichum isolates was obtained from 14 tea cultivars in 86 tea plantations. Phylogenic analysis using the ribosomal internal transcribed spacer, glutamine synthetase gene, Apn2-Mat1-2 intergenic spacer, β-tubulin, actin, calmodulin, and glyceraldehyde-3-phosphate dehydrogenase genes together with morphological characterization revealed three species associated with brown blight of tea; namely, Colletotrichum camelliae (95.6% of all isolates), C. fructicola (3.7%), and C. aenigma (0.7%). This is the first report of C. aenigma in Taiwan. The optimal growth temperatures were 25°C for C. camelliae and 25 and 30°C for C. fructicola and C. aenigma . Although C. fructicola and C. aenigma were more adapted to high temperature, C. camelliae was the most pathogenic across different temperatures. Regardless of whether spore suspensions or mycelial discs were used, significantly larger lesions and higher disease incidences were observed for wounded than for nonwounded inoculation and for the third and fourth leaves than for the fifth leaves. Wounded inoculation of detached third and fourth tea leaves with mycelial discs was found to be a reliable and efficient method for assessing the pathogenicity of Colletotrichum spp. within 4 days. Preventive application of fungicides or biocontrol agents immediately after tea pruning and at a young leaf stage would help control the disease.

The infection of Macrophomina phaseolina often results in a grayish appearance with numerous survival structures, microsclerotia, on the plant surface. Past works have studied the development of fungal survival structures, sclerotia and microsclerotia, in the Leotiomycetes and Sordariomycetes. However, M. phaseolina belongs to the Dothideomycetes, and it remains unclear whether the mechanism of microsclerotia formation remains conserved among these phylogenetic clades. This study applied RNA-sequencing (RNA-Seq) to profile gene expressions at four stages of microsclerotia formation, and the results suggested that reactive oxygen species (ROS)-related functions were significantly different between the microsclerotia stages and the hyphal stage. Microsclerotia formation was reduced in the plates amended with antioxidants such as ascorbic acid, dithiothreitol (DTT), and glutathione. Surprisingly, DTT drastically scavenged H 2 O 2 , but the microsclerotia amount remained similar to the treatment of ascorbic acid and glutathione that both did not completely eliminate H 2 O 2 . This observation suggested the importance of [Formula: see text] over H 2 O 2 in initiating microsclerotia formation. To further validate this hypothesis, the superoxide dismutase 1 (SOD1) inhibitor diethyldithiocarbamate trihydrate (DETC) and H 2 O 2 were tested. The addition of DETC resulted in the accumulation of endogenous [Formula: see text] and more microsclerotia formation, but the treatment of H 2 O 2 did not. The expression of SOD1 genes were also found to be upregulated in the hyphae to the microsclerotia stage, which suggested a higher endogenous [Formula: see text] stress presented in these stages. In summary, this study not only showed that the ROS stimulation remained conserved for initiating microsclerotia formation of M. phaseolina but also highlighted the importance of [Formula: see text] in initiating the hyphal differentiation to microsclerotia formation. IMPORTANCE Reactive oxygen species (ROS) have been proposed as the key stimulus for sclerotia development by studying fungal systems such as Sclerotinia sclerotiorum, and the theory has been adapted for microsclerotia development in Verticillium dahliae and Nomuraea rileyi. While many studies agreed on the association between (micro)sclerotia development and the ROS pathway, which ROS type, superoxide ([Formula: see text]) or hydrogen peroxide (H 2 O 2 ), plays a major role in initiating hyphal differentiation to the (micro)sclerotia formation remains controversial, and literature supporting either [Formula: see text] or H 2 O 2 can be found. This study confirmed the association between ROS and microsclerotia formation for the charcoal rot fungus Macrophomina phaseolina. Moreover, the accumulation of [Formula: see text] but not H 2 O 2 was found to induce higher density of microsclerotia. By integrating transcriptomic and phenotypic assays, this study presented the first conclusive case for M. phaseolina that [Formula: see text] is the main ROS stimulus in determining the amount of microsclerotia formation.

Jabuticaba (Plinia cauliflora (Mart.) Kausel) was originated from Brazil (Lorenzi 2000). The production of jabuticaba is growing globally as its value in the food and pharmaceutical industries (Benvenutti 2021). In August 2019, jabuticaba plants with symptoms of leaf blight were observed in the field at the Meinong of Kaohsiung City, Taiwan. Disease incidence was 40%. Symptoms first presented as small, water-soaked lesions on young leaves, and then dark brown lesions of 1-3 cm in diameter on mature leaves. Six symptomatic leaves were collected from 6 jabuticaba plants for verifying the causal agents. Tissues (5 × 5 mm2) were cut from the margin of symptomatic leaf. Samples were sterilized in 1% sodium hypochlorite for 60 s, rinsed with sterile distilled water three times and then placed in 1% water agar in the dark for 5 days at room temperature. Resultant fungal colonies were purified by subculturing fungal hyphal tips on potato dextrose agar in a growth chamber (28°C, 12 h photoperiod) until fungal conidia appeared. The fungi initially produced white, cottony, aerial mycelium, after which concentric black conidiomata appeared on the plates after 7 days of incubation. The 5-celled conidia were fusiform to ellipsoid, straight to slightly curved, with sizes of 24.00-44.00 µm × 6.00-13.00 µm (avg. size, 32.00 × 9.37 µm, n = 120). The apical and basal cells were hyaline, and 3 median cells were pale brown and versicolorous. Conidia had 2-3 apical appendages and a conical basal cell with a truncate base. Based on the characteristics, which were common among isolates from diseased samples, the causal pathogen was identified as Neopestalotiopsis sp. (Solarte et al. 2018). Internal transcribed spacer (ITS), translation elongation factor 1α (TEF), β-tubulin, and large ribosomal subunit (LSU) DNA sequences were obtained from these isolates and deposited in GenBank (MN723897, ITS; MN813055, TEF; MN813054, β-tubulin; MN860104, LSU). Sequences demonstrated high sequence identity with those of Neopestalotiopsis formicarum ex-type cultures CBS 362.72 (Maharachchikumbura et al. 2014): 99.44% for ITS (KM199358), 99.38% for TEF (KM199517), 98.86% for β-tubulin (KM199455), and 100.00% for LSU (KM116248). The phylogenetic relationship in Neopestalotiopsis species supported the identification of our isolates as N. formicarum. Three independent 3-isolate inoculation experiments were performed to fit Koch's postulates. Surface-sterilized leaves on live plants were punctured with a needle and inoculated with 5 µL of conidial suspension (1 × 105 conidia/mL). Inoculated plants were kept in a growth chamber (25°C, 70% relative humidity) for 7 days. Control plants were inoculated with sterile distilled water and kept under the same conditions. Inoculated leaves developed brown lesions around wounds after seven days. The pathogen was re-isolated from diseased plants, following the steps used for the original procedure, with identical characteristics as the initial isolates. This is the first report of leaf brown blight caused by N. formicarum on jabuticaba in Taiwan. N. formicarum was recently considered as a new threat to jabuticaba (Gualberto et al. 2021). In addition, it has a broad host range on many tropical crops, such as guarana and banana (Gualberto et al. 2021). Neopestalotiopsis spp. have been reported to cause important economic fruit diseases (Gualberto et al. 2021). Therefore, N. formicarum may become the potential risk for fruit production of tropical crops.

Calonectria ilicicola (anamorph: Cylindrocladium parasiticum ) is a soilborne plant-pathogenic fungus with a broad host range, and it can cause red crown rot of soybean and Cylindrocladium black rot of peanut, which has become an emerging threat to crop production worldwide. Limited molecular studies have focused on Calonectria ilicicola and one of the possible difficulties is the lack of genomic resources. This study presents the first high quality and near-completed genome of C. ilicicola , using the Oxford Nanopore GridION sequencing platform. A total of 16 contigs were assembled and the genome of C. ilicicola isolate F018 was estimated to have 11 chromosomes. Currently, the C. ilicicola F018 genome represents the most contiguous assembly, which has the lowest contig number and the highest contig N 50 among all Calonectria genome resources. Putative protein-coding sequences and secretory proteins were estimated to be 17,308 and 1,930 in the C. ilicicola F018 genome, respectively; and the prediction was close to other plant-pathogenic fungi, such as Fusarium species, within the Nectriaceae family. The availability of this high-quality genome resource is expected to facilitate research on fungal biology and genetics of C. ilicicola and to support advanced understanding of pathogen virulence and disease management.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Successive cultivation of fungi on artificial media has been reported to cause the sectorization, which leads to degeneration of developmental phenotype, and virulence. Fusarium oxysporum f. sp. niveum (Fon), the causal agent of watermelon Fusarium wilt, forms degenerated sectors after successive cultivation. In the present research, we demonstrated that subculture with aged mycelia increased the incidence of degenerations. To further investigate the differences between the Fon wild type (sporodochial type, ST) and variants (MT: mycelial type and PT: pionnotal type), developmental phenotypes and pathogenicity to watermelon were examined. Results in variants (PT2, PT3, PT11, and MT6) were different from ST with mycelia growth, conidia production and chlamydospore formation. Virulence of degenerated variants on susceptible watermelon Grand Baby (GB) cultivar was determined after inoculation with Fon variants and Fon ST. In root dipping methods, Fon variants showed no significant differences in disease progress compared with ST. Fon variants showed a significant decrease in disease progression compared with ST through infested soil inoculation. The contrasting results of two inoculation methods suggest that the degenerative changes due to repeated successive cultivation may lead to the loss of pathogen virulence-related factors of the early stage of Fon infection process. Therefore, cell wall-degrading enzymes (CWDEs; cellulase, pectinase, and xylanase) activities of different variants were analyzed. All Fon degenerated variants demonstrated significant decreases of CWDEs activities compared with ST. Additionally, transcript levels of 9 virulence-related genes ( fmk1 , fgb1 , pacC , xlnR , pl1 , rho1 , gas1 , wc1 , and fow1 ) were assessed in normal state. The degenerated variants demonstrated a significantly low level of tested virulence-related gene transcripts except for fmk1 , xlnR, and fow1 . In summary, the degeneration of Fon is triggered with successive subculture through aged mycelia. The degeneration showed significant impacts on virulence to watermelon, which was correlated with the reduction of CWDEs activities and declining expression of a set of virulence-related genes.

Tea (Camellia sinensis (L.) O. Kuntze) is a very popular beverage and cash crop that is widely cultivated in tropical and subtropical areas. In November 2017, diseased tea plants that exhibiting brown blight disease were observed in Guanxi Township of Hsinchu County in Taiwan. In the plantation,15% of tea trees (about 4000 plants) had an average of 20% of the leaves with at least one lesion. The symptoms began as small, water-soaked lesions on young leaves and twigs and later became larger, dark brown, necrotic lesions of 1 to 3 cm in diameter on leaves and 2 to 5 cm in length on twigs. Symptomatic leaf tissue (1 cm2) from five samples per sample) was surface sterilized with 1% NaClO (from commercial bleach, Clorox) for 1 min, washed with sterilized water 3 times, plated onto potato dextrose agar (PDA), and incubated under 12h/12h cycles of light and darkness at 25°C until sporulation to determine the causal agent. A fungus was consistently isolated from symptomatic leaf samples (80% isolation rate). The fungus initially produced white-to-gray fluffy aerial hyphae, which subsequently exhibited dark pigmentation. Acervuli and setae were absent. The conidia were hyaline, aseptate, smooth-walled, and cylindrical with obtuse to slightly rounded ends, with sizes of 12.10 to 16.02 × 3.58 to 4.91 (average 13.77 × 4.05, n = 30) μm. The majority had two rounded guttules. The appressoria were brown to dark brown, ovoid and slightly obtuse at the tip in shape, had lengths ranging from 3.59 to 10.31 μm (with an average of 7.18 μm, n = 30), and had diameters of 3.14 to 6.43 μm (with an average of 5.10 μm, n = 30). Morphological characteristics matched the descriptions of Colletotrichum fructicola (Liu et al. 2015; Fuentes-Aragón et al. 2018). The internal transcribed spacer of nuclear ribosomal DNA (ITS), actin (ACT), chitin synthase (CHS-1), and Apn2-Mat1-2 intergenic spacer and partial mating-type Mat1-2 gene (ApMAT) sequences of the isolates were obtained to confirm this identification. The sequences showed close identity with those of C. fructicola ex-type cultures ICMP18581 and CBS 130416 (Weir et al. 2012) of 99.65% for the ITS (JX010165), 99.29% for the ACT (JX009501), and 100.00% for the CHS-1 (JX009866), as well as close identity with the other ex-type culture LF506 (Liu et al. 2015) of 99.59% for the ApMat (KJ954567), supporting the isolate's identification as C. fructicola. The sequences were deposited in GenBank, with the following accession Nos.: MN608177 (ITS), MN393175 (ACT), MT087546 (CHS-1), and MT087542 (ApMAT). Based on morphology and DNA sequence analysis, the associated fungus was identified as C. fructicola. Pathogenicity tests were performed next according to the procedures described in Chen et al. (2017). Healthy leaves on tea plants (Ca. sinensis 'Chin-shin Oolong') were wounded by pinpricking in the middle of each counterpart and inoculated with conidial suspension (1 × 107 conidia/ml, 10 μl). Both non-wounded and wounded healthy leaves were inoculated with the conidial suspension and sterile distilled water (a water control). The tea plants were covered with plastic bags to maintain high relative humidity for two days. One week after inoculation, anthracnose was observed on 40% of inoculated leaves, whereas all the control leaves remained healthy. The fungus was re-isolated from the diseased plants, and identified as C. fructicola by resequencing of the four genes. To the best of our knowledge, this is the first report of anthracnose caused by C. fructicola on tea in Taiwan although the pathogen has been present in China and Indonesia (Wang et al. 2016; Shi et al. 2017; Farr and Rossman, 2020).

Core-shell InSb-SiO(2) nanoballs/microballs were synthesized on a Si substrate by carbonthermal reactions at a temperature of 900 °C. High-resolution transmission microscopy (HRTEM) images revealed that the surfaces of the InSb nanoballs/microballs were covered by amorphous SiO(2) layers. On the basis of our theoretical calculation, the thermal expansion coefficient (TEC) of the InSb crystals is ten times higher than that of the SiO(2) shell. Therefore, the SiO(2) serves as a constraining shell for the InSb core so that the compressive stress of ∼-94 MPa can accumulate in the InSb core while a tensile stress of 196 MPa forms in the SiO(2) shell. The thermal excitation accumulated compressive stress in the InSb core, causing a partial structural phase transition from a cubic zinc-blende structure to a hexagonal wurtzite structure. Many lattice defects, such as stacking faults and Moiré fringes, have been observed on the surface of the InSb core. In situ temperature-dependent XRD patterns showed that a reversible InSb hexagonal (002) peak appeared and disappeared as the temperature increased and decreased at a transit point of 200 °C, respectively. As the temperature increased, the XRD diffraction peaks of the InSb wurtzite phase shifted significantly to lower angles because of the formation of compressive stress in the InSb nanoballs. The pressure-induced partial structural phase transitions of the nanostructured InSb occurred at -94 MPa of the compressive stress. This is the first report of this value, which is the lowest value in the pressure-induced phase transition of the nanostructure InSb from the cubic zinc-blende structure to the hexagonal wurtzite structure.

Objective: To diagnose and distinguish the swell in the infantile scrotum or testicle by the high frequency ultrasonoscopy so as to find the acoustic imaging characters and clinical application value., Methods: With the high frequency linear-array probe used, the multi-section and two-side contrast scanning checks were conducted on 46 cases of the infantile scrotum or testicle swell and position with abnormal physical signs. The structures of scrotum, testicle, epididymis and spermatic cord as well as the positions, sizes, forms and interior echoes were observed. When it was necessary, the position should be changed, bladder filled up and the groin and two sides of bladder scanned., Results: A total of 45 cases of infantile scrotum or testicle disease were diagnosed by ultrasound, which accorded with the clinical behavior and operation result while 1 case of one-side cryptorchid was not found., Conclusion: It is quick and accurate to diagnose the infantile scrotum or testicle disease by high frequency ultrasonoscopy.

We have identified two azoospermic men with r(Y) in 312 infertile men presenting with non-obstructive azoospermia or oligozoospermia. Their karyotypes were 45,X [9]/46,X, r(Y)(p11q11) [11] (case 1), and 46,X,r(Y)(p11q11) (case 2), respectively. In both cases, the Yp breakpoints were located within the pseudoautosomal region. Both cases had extensive deletions of azoospermia factors (AZFs). Case 1 also had deletion of the putative growth controlling gene (GCY) and the Yq breakpoint was located between sY741 and USP9Y. The Yq breakpoint was located between sY105 and sY109 in case 2. Both cases did not have Turner stigmata except short stature in case 1. By a combination of cytogenetic and molecular genetic tools, we showed r(Y) arose from breakage in both arms of the chromosome with subsequent fusion of two broken ends of the centric fragment to form a continuous ring. Spermatogenic defects in men with r(Y) may result from deletion of Y-linked AZFs combined with synaptic failure., (Copyright 2004 Wiley-Liss, Inc.)