α1-COP modulates plasmodesmata function through sphingolipid enzyme regulation.

Callose, a ß-1,3-glucan plant cell wall polymer, regulates symplasmic channel size at plasmodesmata (PD) and plays a crucial role in a variety of plant processes. However, elucidating the molecular mechanism of PD callose homeostasis is limited. We screened and identified an Arabidopsis mutant plant with excessive callose deposition at PD and found that the mutated gene was α1-COP, a member of the coat protein I (COPI) coatomer complex. We report that loss of function of α1-COP elevates the callose accumulation at PD by affecting subcellular protein localization of callose degradation enzyme PdBG2. This process is linked to the functions of ERH1, an inositol phosphoryl ceramide synthase, and glucosylceramide synthase through physical interactions with the α1-COP protein. Additionally, the loss of function of α1-COP alters the subcellular localization of ERH1 and GCS proteins, resulting in a reduction of GlcCers and GlcHCers molecules, which are key sphingolipid (SL) species for lipid raft formation. Our findings suggest that α1-COP protein, together with SL modifiers controlling lipid raft compositions, regulates the subcellular localization of GPI-anchored PDBG2 proteins, and hence the callose turnover at PD and symplasmic movement of biomolecules. Our findings provide the first key clue to link the COPI-mediated intracellular trafficking pathway to the callose-mediated intercellular signaling pathway through PD.

Antifungal Properties of Streptomyces bacillaris S8 for Biological Control Applications.

Soybean (Glycine max), a crucial global crop, experiences yearly yield reduction due to diseases such as anthracnose (Colletotrichum truncatum) and root rot (Fusarium spp.). The use of fungicides, which have traditionally been employed to control these phytopathogens, is now facing challenges due to the emergence of fungicide-resistant strains. Streptomyces bacillaris S8 strain S8 is previously known to produce valinomycin t through a nonribosomal peptide synthetase (NRPS) pathway. The objective of this study was to evaluate the antifungal activity of S. bacillaris S8 against C. truncatum and Fusarium sp., assessing its efficacy against soybean pathogens. The results indicate that strain S8 effectively controlled both above-ground and underground soybean diseases, using the NRPS and NRPS-related compound, suggesting its potential as a biological control in plant-microbe interactions. These findings underscore the pivotal role of the stain S8 in fostering healthy soybean microbial communities and emphasize the significance of microbiota structure studies in unveiling potent biocontrol agents.

In Vivo Human Lacrimal Gland Imaging Using an Ultrasound Biomicroscopy.

PURPOSE: In the present study, we introduce human lacrimal gland imaging using an ultrasound biomicroscopy (UBM) with a soft cover and show their findings. METHODS: The representative UBM findings of palpebral lobes in seven subjects (four with non-Sjögren dry eye syndrome, one with Sjögren syndrome, and two healthy subjects) were described in this study. To prolapse the palpebral lobe, the examiner pulled the temporal part of the upper eyelid in the superotemporal direction and directed the subject to look in the inferonasal direction. We scanned the palpebral lobes longitudinally and transversely using UBM. We used an Aviso UBM with a 50 MHz linear probe and ClearScan. RESULTS: In UBM of two healthy subjects, the echogenicity of the lacrimal gland was lower than that of the sclera and homogeneous. But the parenchyma of a patient with Sjögren dry eye syndrome was quite inhomogeneous compared to the healthy subjects. In two patients with dry eye syndrome, we were able to observe some lobules in the parenchyma. We could find excretory ducts running parallel at the surface of the longitudinal section in some subjects. In the longitudinal UBM scan of a subject, we observed a tubular structure at a depth of 1,500 µm that was considered a blood vessel. It ran from the superonasal to the inferotemporal direction. In a subject, we observed a large cyst beneath the conjunctiva. CONCLUSIONS: Lacrimal gland imaging using UBM has both advantages of optical coherence tomography and sonography, and could be useful for evaluating dry eye syndrome.

Ocular parameters associated with visual performance of enhanced monofocal intraocular lens.

BACKGROUND: An enhanced monofocal intraocular lenses (IOLs) (Tecnis Eyhance ICB00 and Tecnis Eyhance Toric DIU) has been developed to enhance intermediate vision while avoiding the disadvantages of multifocal IOLs. Although many studies have demonstrated the improvement of intermediate visual acuity with enhanced monofocal IOLs, it is not known specifically for which patients these IOLs should be recommended or avoided. In this study, we aim to find out which ocular parameters affect vision performance and photic phenomenon of ICB00 or DIU at different distances. METHODS: Patients who underwent cataract surgery with ICB00 or DIU, performed by a single surgeon, were included. Before surgery, the patients' age, gender, axial length, anterior chamber depth, spherical aberration Z (4,0), vertical coma, horizontal coma, angle kappa (κ), angle alpha (α), and other ocular parameters were investigated. One month after surgery, uncorrected near visual acuity (UNVA at 40 cm), uncorrected intermediate visual acuity (UIVA at 66 cm), uncorrected distance logMAR visual acuity (UDVA), IOL decentration, and quality of vision (QoV) questionnaires were conducted. RESULTS: A total of 43 patients (58 eyes) were included. The results of the univariate linear regression analyses showed a negative correlation between spherical aberration and logMAR UNVA and UIVA (p = 0.003, ß=-0.51 and p = 0.018, ß=-0.23, respectively) and a positive correlation between angle α and logMAR UIVA (p = 0.036, ß = 0.19). Deeper anterior chamber depth (ACD) was associated with poorer total QoV (p = 0.018, ß = 14.43), particularly in glare, halo, blur, and fluctuation perception. A higher degree of IOL decentration tended to decrease UNVA and UIVA (Pearson correlation coefficient, r = 0.336 and r = 0.221, respectively); however, no significant effect was observed on UDVA (Pearson correlation coefficient, r = 0.042). CONCLUSIONS: In enhanced monofocal IOLs, a higher level of spherical aberration is associated with better performance in UNVA and UIVA, whereas a larger angle α has a negative impact. A deeper ACD negatively affects the QoV.

Categorization of Meibomian Gland Dysfunction Using Lipid Layer Thickness and Meibomian Gland Dropout in Dry Eye Patients: A Retrospective Study.

PURPOSE: In the present study, we determined the prevalence of obstructive meibomian gland dysfunction (MGD), hyposecretory MGD, grossly normal MG, and hypersecretory MGD in patients with dry eye syndrome using lipid layer thickness (LLT) and MG dropout. METHODS: Eighty-eight patients with dry eye syndrome were included in the study. Patients were categorized into four groups according to the LLT and weighted total meiboscore. The proportion of patients in each group was calculated. The age, sex, Ocular Surface Disease Index, LLT, Schirmer, tear film breakup time, cornea stain, weighted total meiboscore, expressibility, and quality of meibum were compared between the four groups. RESULTS: Fifteen eyes (17.0%) had obstructive MGD, two eyes (2.3%) had hyposecretory MGD, 40 eyes (45.5%) had grossly normal MG, and 17 eyes (19.3%) had hypersecretory MGD. The obstructive MGD group was younger than the grossly normal MG group. In obstructive MGD, the ratio of men to women was higher than that of the other groups. However, Ocular Surface Disease Index, Schirmer, tear film breakup time, and corneal stain did not show statistically significant differences between the four groups. The meibum expressibility of the hyposecretoy MGD group was worse than those of the other groups. The meibum expressibility of the hyposecretoy MGD group was poor than those of the obstructive and hypersecretory MGD group. CONCLUSIONS: This categorization was expected to help determine the best treatment method for dry eye syndrome, according to the MG status.

Unraveling the Role of Cytochrome P450 as a Key Regulator Lantipeptide Production in Streptomyces globisporus.

The aim of this study was to investigate the regulation of lantipeptide production in Streptomyces globisporus SP6C4, which produces the novel antifungal lantipeptides conprimycin and grisin, and to identify the role of cytochrome P450 (P450) in tis regulation. To investigate the regulation of lantipeptide production, we created gene deletion mutants, including ΔP450, ΔtsrD, ΔlanM, ΔP450ΔtsrD, and ΔP450ΔlanM. These mutants were characterized in terms of their morphology, sporulation, attachment, and antifungal activity against Fusarium oxysporum. The gene deletion mutants showed distinct characteristics compared to the wild-type strain. Among them, the ΔP450ΔlanM double mutant exhibited a recovery of antifungal activity against F. oxysporum, indicating that P450 plays a significant role in regulating lantipeptide production in S. globisporus SP6C4. Our findings highlight the significant role of P450 in the regulation of lantipeptide production and morphological processes in S. globisporus. The results suggest a potential link between P450-mediated metabolic pathways and the regulation of growth and secondary metabolism in SP6C4, thereby highlighting P450 as a putative target for the development of new antifungal agents.

Mycobiota community and fungal species response to development stage and fire blight disease in apples.

Fire blight disease, caused by the bacterial pathogen Erwinia amylovora, has been a significant concern for over 50 countries worldwide. The efficacy of chemical pesticides currently available for disease control is limited. To address this issue, research is being conducted to explore environmentally friendly control methods, particularly biological control using beneficial microorganisms. However, there is limited research on the apple microbiota community and minimal research has been conducted on fungal communities that may exhibit reliable performance in apple trees. Therefore, our objective was to analyze the fungal communities present in apples at different developmental stages and in different tissues, aiming to identify potential biological control agents for fire blight disease. Our findings indicate that the fungal communities present in apple buds, flowers and leaves play an important role in inhibiting the invasion of E. amylovora. Specifically, we propose GS11 and Lipomyces starkeyi as potential keystone taxa that respond to fire blight disease. These findings provide insights into the continuity and discontinuity of fungal community structure in different developmental stages of apples and offer predictions for potential biological control agents for fire blight disease.

Analysis of Endophytic Bacterial Communities and Investigation of Core Taxa in Apple Trees.

ire blight disease, caused by Erwinia amylovora, is a devastating affliction in apple cultivation worldwide. Chemical pesticides have exhibited limited effectiveness in controlling the disease, and biological control options for treating fruit trees are limited. Therefore, a relatively large-scale survey is necessary to develop microbial agents for apple trees. Here we collected healthy apple trees from across the country to identify common and core bacterial taxa. We analyzed the endophytic bacterial communities in leaves and twigs and discovered that the twig bacterial communities were more conserved than those in the leaves, regardless of the origin of the sample. This finding indicates that specific endophytic taxa are consistently present in healthy apple trees and may be involved in vital functions such as disease prevention and growth. Furthermore, we compared the community metabolite pathway expression rates of these endophyte communities with those of E. amylovora infected apple trees and discovered that the endophyte communities in healthy apple trees not only had similar community structures but also similar metabolite pathway expression rates. Additionally, Pseudomonas and Methylobacterium-Methylorobrum were the dominant taxa in all healthy apple trees. Our findings provide valuable insights into the potential roles of endophytes in healthy apple trees and inform the development of strategies for enhancing apple growth and resilience. Moreover, the similarity in cluster structure and pathway analysis between healthy orchards was mutually reinforcing, demonstrating the power of microbiome analysis as a tool for identifying factors that contribute to plant health.

Dynamics of Bacterial Communities by Apple Tissue: Implications for Apple Health.

Herein, we explored the potential of the apple's core microbiota for biological control of Erwinia amylovora, which causes fire blight disease, and analyzed the structure of the apple's bacterial community across different tissues and seasons. Network analysis results showed distinct differences in bacterial community composition between the endosphere and rhizosphere of healthy apples, and eight taxa were identified as negatively correlated with E. amylovora, indicating their potential key role in a new control strategy against the pathogen. This study highlights the critical role of the apple's bacterial community in disease control and provides a new direction for future research in apple production. In addition, the findings suggest that using the composition of the apple's core taxa as a biological control strategy could be an effective alternative to traditional chemical control methods, which have been proven futile and environmentally harmful.

Transition from Ginseng Root Rot Disease-Conducive Soil to -Suppressive Soil Mediated by Pseudomonadaceae.

Ginseng is a popular medicinal herb with established therapeutic effects such as cardiovascular disease prevention, anticancer effects, and anti-inflammatory effects. However, the slow growth of ginseng due to soilborne pathogens has been a challenge for establishing new plantations. In this study, we investigated root rot disease associated with the microbiota in a ginseng monoculture model system. Our results showed that a collapse of the early microbiota community inhibiting root rot disease was observed before the disease became severe, and nitrogen fixation was necessary to support the initial microbiota community structure. Furthermore, changes in the nitrogen composition were essential for the suppression of pathogen activity in early monoculture soils. We hypothesize that Pseudomonadaceae, a population built up by aspartic acid, can inhibit the occurrence of root rot disease in ginseng and that specific management practices that maintain a healthy microbiome can be implemented to prevent and mitigate the disease. Our findings provide insights into the potential use of specific members of the microbiota for controlling root rot disease in ginseng cultivation. IMPORTANCE Understanding the initial soil microbiota and community shifts in a monoculture system is critical for developing disease-suppressive soils for crop production. The lack of resistance genes against soilborne pathogens in plants highlights the need for effective management strategies. Our investigation of root rot disease and initial microbiota community shifts in a ginseng monoculture model system provides valuable insight into the development of conducive soil into specific suppressive soil. With a thorough understanding of the microbiota in disease-conducive soil, we can work toward the development of disease-suppressive soil to prevent outbreaks and ensure sustainable crop production.

Endophytic Streptomyces population induced by L-glutamic acid enhances plant resilience to abiotic stresses in tomato.

Endophyte bacteria, which colonize plants including roots, stem, flower, and fruit, it can derive their nutrients from the host, are recognized for their mutualistic relationship with the host plant. They play a critical role in promoting host growth and modulating abiotic stress. Carbon and nitrogen have a significant impact on bacterial population and secondary metabolite production, which are highly specific in various categories such as bacterial growth regulation, anti-compounds production. Application of L-glutamic acid can significantly enhance Streptomyces globisporus population buildup in plants. However, the effectiveness of this population buildup against abiotic stresses such as salinity and drought has not been investigated. Therefore, in this study, we tested the bacteria and their prebiotic activity against salinity and drought stress in tomato plants. Three different amino acids were treated on the tomato plants, and it was observed that L-asparagine and L-proline had a negative effect on plant growth and phenotype, while L-glutamic acid promoted plant growth and increased bacteria population density. The bacteria were found to colonize the rhizosphere and root endosphere, with colonization being promoted by L-glutamic acid. Additionally, Streptomyces was found to have plant growth promotion effects and provided protection against abiotic stresses. Interestingly, L-glutamic acid reduced the damage caused by salinity stress, but not drought stress. These findings suggest that L-glutamic acid plays a role in providing tolerance to salinity stress with the core microbiota, thus the current study demonstrated their prebiotic activity in the agriculture system.

Variations in Kiwifruit Microbiota across Cultivars and Tissues during Developmental Stages.

The plant microbiota plays a crucial role in promoting plant health by facilitating the nutrient acquisition, abiotic stress tolerance, biotic stress resilience, and host immune regulation. Despite decades of research efforts, the precise relationship and function between plants and microorganisms remain unclear. Kiwifruit (Actinidia spp.) is a widely cultivated horticultural crop known for its high vitamin C, potassium, and phytochemical content. In this study, we investigated the microbial communities of kiwifruit across different cultivars (cvs. Deliwoong and Sweetgold) and tissues at various developmental stages. Our results showed that the microbiota community similarity was confirmed between the cultivars using principal coordinates analysis. Network analysis using both degree and eigenvector centrality indicated similar network forms between the cultivars. Furthermore, Streptomycetaceae was identified in the endosphere of cv. Deliwoong by analyzing amplicon sequence variants corresponding to tissues with an eigenvector centrality value of 0.6 or higher. Our findings provide a foundation for maintaining kiwifruit health through the analysis of its microbial community.

Roads to Construct and Re-build Plant Microbiota Community.

Plant microbiota has influenced plant growth and physiology significantly. Plant and plant-associated microbes have flexible interactions that respond to changes in environmental conditions. These interactions can be adjusted to suit the requirements of the microbial community or the host physiology. In addition, it can be modified to suit microbiota structure or fixed by the host condition. However, no technology is realized yet to control mechanically manipulated plant microbiota structure. Here, we review step-by-step plant-associated microbial partnership from plant growth-promoting rhizobacteria to the microbiota structural modulation. Glutamic acid enriched the population of Streptomyces, a specific taxon in anthosphere microbiota community. Additionally, the population density of the microbes in the rhizosphere was also a positive response to glutamic acid treatment. Although many types of research are conducted on the structural revealing of plant microbiota, these concepts need to be further understood as to how the plant microbiota clusters are controlled or modulated at the community level. This review suggests that the intrinsic level of glutamic acid in planta is associated with the microbiota composition that the external supply of the biostimulant can modulate.

Microbiota Communities of Healthy and Bacterial Pustule Diseased Soybean.

Soybean is an important source of protein and for a wide range of agricultural, food, and industrial applications. Soybean is being affected by Xanthomonas citri pv. glycines, a causal pathogen of bacterial pustule disease, result in a reduction in yield and quality. Diverse microbial communities of plants are involved in various plant stresses is known. Therefore, we designed to investigate the microbial community differentiation depending on the infection of X. citri pv. glycines. The microbial community's abundance, diversity, and similarity showed a difference between infected and non-infected soybean. Microbiota community analysis, excluding X. citri pv. glycines, revealed that Pseudomonas spp. would increase the population of the infected soybean. Results of DESeq analyses suggested that energy metabolism, secondary metabolite, and TCA cycle metabolism were actively diverse in the non-infected soybeans. Additionally, Streptomyces bacillaris S8, an endophyte microbiota member, was nominated as a key microbe in the healthy soybeans. Genome analysis of S. bacillaris S8 presented that salinomycin may be the critical antibacterial metabolite. Our findings on the composition of soybean microbiota communities and the key strain information will contribute to developing biological control strategies against X. citri pv. glycines.

Glutamic acid reshapes the plant microbiota to protect plants against pathogens.

BACKGROUND: Plants in nature interact with other species, among which are mutualistic microorganisms that affect plant health. The co-existence of microbial symbionts with the host contributes to host fitness in a natural context. In turn, the composition of the plant microbiota responds to the environment and the state of the host, raising the possibility that it can be engineered to benefit the plant. However, technology for engineering the structure of the plant microbiome is not yet available. RESULTS: The loss of diversity and reduction in population density of Streptomyces globisporus SP6C4, a core microbe, was observed coincident with the aging of strawberry plants. Here, we show that glutamic acid reshapes the plant microbial community and enriches populations of Streptomyces, a functional core microbe in the strawberry anthosphere. Similarly, in the tomato rhizosphere, treatment with glutamic acid increased the population sizes of Streptomyces as well as those of Bacillaceae and Burkholderiaceae. At the same time, diseases caused by species of Botrytis and Fusarium were significantly reduced in both habitats. We suggest that glutamic acid directly modulates the composition of the microbiome community. CONCLUSIONS: Much is known about the structure of plant-associated microbial communities, but less is understood about how the community composition and complexity are controlled. Our results demonstrate that the intrinsic level of glutamic acid in planta is associated with the composition of the microbiota, which can be modulated by an external supply of a biostimulant. Video Abstract.

Development of a novel multifocal lens using a polarization directed flat lens: possible candidate for a multifocal intraocular lens.

BACKGROUND: A polarization-directed flat (PDF) lens acts as a converging lens with a focal length (f) > 0 and a diverging lens with f < 0, depending on the polarization state of the incidental light. To produce a multifocal lens with two focal lengths, a PDF and a converging lens having shorter focal length were combined. In this study, we tested a bifocal PDF to determine its potential as a new multifocal intraocular lens (IOL). METHODS: Constructed a multifocal lens with a PDF lens (f = +/- 100 mm) and a converging lens (f = + 25 mm). In an optical bench test, we measured the defocus curve to test the multifocal function. The multifocal function and optical quality of the lens in various situations were tested. An Early Treatment Diabetic Retinopathy Study (ETDRS) chart as a near target and a building as a distant target were photographed using a digital single-lens reflex (DSLR) camera. Both lenses (multifocal and monofocal) were tested under the same conditions. RESULTS: For the 0 D and - 20 D focal points, the multifocal lens showed sharp images in the optical bench test. In the DSLR test using the multifocal lens, the building appeared slightly blurry compared with the results using the monofocal lens. With the multifocal lens, the ETDRS chart's images became blurry as the ETDRS chart's distance decreased, but became very clear again at a certain position. CONCLUSIONS: We confirmed the multifocal function of the multifocal lens using a PDF lens. This lens can be used as a multifocal IOL in the future.

Association between diabetes status and subsequent onset of glaucoma in postmenopausal women.

The purpose of this study was to analyze the risk of glaucoma based on diabetes status using a large nationwide longitudinal cohort of postmenopausal women. This study included 1,372,240 postmenopausal women aged ≥ 40 years who underwent National Health Screening Program in 2009. Subjects were classified into the following 5 categories based on diabetes status: no diabetes, impaired fasting glucose (IFG), new onset diabetes, diabetes treated with oral hypoglycemic medication, and diabetes treated with insulin. Subjects were followed from 2005 through 2018, and hazard ratios of glaucoma onset were calculated for each group. Subgroup analyses of subjects stratified by age, smoking, drinking, hypertension, and dyslipidemia were performed. During the follow up period, 42,058 subjects developed glaucoma. The adjusted hazard ratio was 1.061 (95% CI, 1.036-1.086) in the IFG group, 1.151 (95% CI, 1.086-1.220) in the new onset diabetes group, 1.449 (95% CI, 1.406-1.493) in the diabetes treated with oral hypoglycemic medication group, and 1.884(95% CI, 1.777-1.999) in the diabetes treated with insulin group compared to the no diabetes group. The results were consistent in subgroup analyses after stratifying by age, lifestyle factors (smoking and drinking), and comorbidities (hypertension and dyslipidemia). Diabetes status is associated with increased risk of glaucoma development in postmenopausal women.

A Genome-Wide Analysis of Antibiotic Producing Genes in Streptomyces globisporus SP6C4.

Soil is the major source of plant-associated microbes. Several fungal and bacterial species live within plant tissues. Actinomycetes are well known for producing a variety of antibiotics, and they contribute to improving plant health. In our previous report, Streptomyces globisporus SP6C4 colonized plant tissues and was able to move to other tissues from the initially colonized ones. This strain has excellent antifungal and antibacterial activities and provides a suppressive effect upon various plant diseases. Here, we report the genome-wide analysis of antibiotic producing genes in S. globisporus SP6C4. A total of 15 secondary metabolite biosynthetic gene clusters were predicted using antiSMASH. We used the CRISPR/Cas9 mutagenesis system, and each biosynthetic gene was predicted via protein basic local alignment search tool (BLAST) and rapid annotation using subsystems technology (RAST) server. Three gene clusters were shown to exhibit antifungal or antibacterial activity, viz. cluster 16 (lasso peptide), cluster 17 (thiopeptide-lantipeptide), and cluster 20 (lantipeptide). The results of the current study showed that SP6C4 has a variety of antimicrobial activities, and this strain is beneficial in agriculture.

Comparison of Bacterial Community of Healthy and Erwinia amylovora Infected Apples.

Fire blight disease, caused by Erwinia amylovora, could damage rosaceous plants such as apples, pears, and raspberries. In this study, we designed to understand how E. amylovora affected other bacterial communities on apple rhizosphere; twig and fruit endosphere; and leaf, and fruit episphere. Limited studies on the understanding of the microbial community of apples and changes the community structure by occurrence of the fire blight disease were conducted. As result of these experiments, the infected trees had low species richness and operational taxonomic unit diversity when compared to healthy trees. Rhizospheric bacterial communities were stable regardless of infection. But the communities in endosphere and episphere were significanlty affected by E. amylovora infection. We also found that several metabolic pathways differ significantly between infected and healthy trees. In particular, we observed differences in sugar metabolites. The finding provides that sucrose metabolites are important for colonization of E. amylovora in host tissue. Our results provide fundamental information on the microbial community structures between E. amylovora infected and uninfected trees, which will contribute to developing novel control strategies for the fire blight disease.