Small Additives Make Big Differences: A Review on Advanced Additives for High-Performance Solid-State Li Metal Batteries.

Solid-state lithium (Li) metal batteries, represent a significant advancement in energy storage technology, offering higher energy densities and enhanced safety over traditional Li-ion batteries. However, solid-state electrolytes (SSEs) face critical challenges such as lower ionic conductivity, poor stability at the electrode-electrolyte interface, and dendrite formation, potentially leading to short circuits and battery failure. The introduction of additives into SSEs has emerged as a transformative approach to address these challenges. A small amount of additives, encompassing a range from inorganic and organic materials to nanostructures, effectively improve ionic conductivity, drawing it nearer to that of their liquid counterparts, and strengthen mechanical properties to prevent cracking of SSEs and maintain stable interfaces. Importantly, they also play a critical role in inhibiting the growth of dendritic Li, thereby enhancing the safety and extending the lifespan of the batteries. In this review, we comprehensively explore the wide variety of additives that have been investigated, emphasizing how they can be effectively incorporated into SSEs. By dissecting the operational mechanisms of these additives, the review hopes to provide valuable insights that can help researchers in developing more effective SSEs, leading to the creation of more efficient and reliable solid-state Li metal batteries. This article is protected by copyright. All rights reserved.

Genotyping-by-Sequencing Analysis Reveals Associations between Agronomic and Oil Traits in Gamma Ray-Derived Mutant Rapeseed (Brassica napus L.).

Rapeseed (Brassica napus L.) holds significant commercial value as one of the leading oil crops, with its agronomic features and oil quality being crucial determinants. In this investigation, 73,226 single nucleotide polymorphisms (SNPs) across 95 rapeseed mutant lines induced by gamma rays, alongside the original cultivar ('Tamra'), using genotyping-by-sequencing (GBS) analysis were examined. This study encompassed gene ontology (GO) analysis and a genomewide association study (GWAS), thereby concentrating on agronomic traits (e.g., plant height, ear length, thousand-seed weight, and seed yield) and oil traits (including fatty acid composition and crude fat content). The GO analysis unveiled a multitude of genes with SNP variations associated with cellular processes, intracellular anatomical structures, and organic cyclic compound binding. Through GWAS, we detected 320 significant SNPs linked to both agronomic (104 SNPs) and oil traits (216 SNPs). Notably, two novel candidate genes, Bna.A05p02350D (SFGH) and Bna.C02p22490D (MDN1), are implicated in thousand-seed weight regulation. Additionally, Bna.C03p14350D (EXO70) and Bna.A09p05630D (PI4Kα1) emerged as novel candidate genes associated with erucic acid and crude fat content, respectively. These findings carry implications for identifying superior genotypes for the development of new cultivars. Association studies offer a cost-effective means of screening mutants and selecting elite rapeseed breeding lines, thereby enhancing the commercial viability of this pivotal oil crop.

METTL3-Mediated N6-methyladenosine mRNA Modification and cGAS-STING Pathway Activity in Kidney Fibrosis.

BACKGROUND: Chemical modifications on RNA profoundly impact RNA function and regulation. m6A, the most abundant RNA modification in eukaryotes, plays a pivotal role in diverse cellular processes and disease mechanisms. However, its importance is understudied in human chronic kidney disease (CKD) samples regarding its influence on pathological mechanisms. METHODS: LC-MS/MS and Methylated RNA Immunoprecipitation (MeRIP) sequencing were utilized to examine alterations in m6A levels and patterns in CKD samples. Overexpression of the m6A writer METTL3 in cultured kidney tubular cells was performed to confirm the impact of m6A in tubular cells and explore the biological functions of m6A modification on target genes. Additionally, tubule-specific deletion of Mettl3 (Ksp-Cre Mettl3f/f) mice and the use of anti-sense oligonucleotides inhibiting Mettl3 expression were utilized to reduce m6A modification in an animal kidney disease model. RESULTS: By examining 127 human CKD samples, we observed a significant increase in m6A modification and METTL3 expression in diseased kidneys. Epitranscriptomic analysis unveiled an enrichment of m6A modifications in transcripts associated with the activation of inflammatory signaling pathways, particularly the cGAS-STING pathway. m6A hypermethylation increased mRNA stability in cGAS and STING1, as well as elevated the expression of key proteins within the cGAS-STING pathway. Both the tubule-specific deletion of Mettl3 and the use of anti-sense oligonucleotides to inhibit Mettl3 expression protected mice from inflammation, reduced cytokine expression, decreased immune cell recruitment, and attenuated kidney fibrosis. CONCLUSIONS: Our research revealed heightened METTL3-mediated m6A modification in fibrotic kidneys, particularly enriching the cGAS-STING pathway. This hypermethylation increased mRNA stability for cGAS and STING1, leading to sterile inflammation and fibrosis.

Instantaneous Thermal Energy for Swift Synthesis of Single-Atom Catalysts for Unparalleled Performance in Metal-Air Batteries and Fuel Cells.

Based on experimental and computational evidence, phthalocyanine (Pc) compounds in the form of quaternary-bound metal-nitrogen (N) atoms are the most effective catalysts for oxygen reduction reaction (ORR). However, the heat treatment process used in their synthesis may compromise the ideal structure, causing the agglomeration of transition metals. To overcome this issue, a novel method is developed for synthesizing iron (Fe) single-atom catalysts with ideal structures supported by thermally exfoliated graphene oxide (GO). This is achieved through a short heat treatment of only 2.5 min involving FePc and N, N-dimethylformamide in the presence of GO. According to the synthesis mechanism revealed by this study, carbon monoxide acts as a strong linker between the single Fe atoms and graphene. It facilitates the formation of a structure containing oxygen species between FeN4 and graphene, which provides high activity and stability for the ORR. These catalysts possess an enormous number of active sites and exhibit enhanced activity toward the alkaline ORR. They demonstrate excellent performance when applied to real electrochemical devices, such as zinc-air batteries and anion exchange membrane fuel cells. It is expected that the instantaneous heat treatment method developed in this study will aid in the development of high-performing single-atom catalysts.

Assessment of Attractant Combinations for the Management of Red Palm Weevils (Rhynchophorus ferrugineus) in the United Arab Emirates.

This study examined red palm weevil ecology in the United Arab Emirates to develop effective food baits, pheromone, and eco-friendly trapping methods. Three phases of investigation were conducted (from June to December 2023) on date palm farms in Ras Al Khaimah and Abu Dhabi. The first two phases, each 15 days long, were conducted in Ras Al Khaimah, whereas the third phase, 18 days long, was conducted in Abu Dhabi. Chemical attractants, such as existing pheromones and ethyl acetate, a newly synthesized ferruginol pheromone, and food baits, such as original dates, date paste, coconut water, and date palm syrup, were used to attract the weevils. Multi-funnel traps containing various attractant mixes were tested. The main activity of the red palm weevils was observed from 3:00 to 6:00 a.m., with 85.72 ± 3.39% being captured during this period, coinciding with cooler temperatures. When pheromones were added to the food bait, the capture rate increased by 6.95 ± 1.81 times. Combining food bait, ethyl acetate, and pheromones improved the capture rates by 3.14 ± 0.69 times compared to pheromones alone. The newly synthesized pheromone achieved capture rates 2.69 ± 0.07 times higher than those of the commercially available pheromone, confirming its suitability as a red palm weevil attractant.

Chromatin accessibility analysis and architectural profiling of human kidneys reveal key cell types and a regulator of diabetic kidney disease.

Diabetes is the leading cause of kidney disease that progresses to kidney failure. However, the key molecular and cellular pathways involved in diabetic kidney disease (DKD) pathogenesis are largely unknown. Here, we performed a comparative analysis of adult human kidneys by examining cell type-specific chromatin accessibility by single-nucleus ATAC-seq (snATAC-seq) and analyzing three-dimensional chromatin architecture via high-throughput chromosome conformation capture (Hi-C method) of paired samples. We mapped the cell type-specific and DKD-specific open chromatin landscape and found that genetic variants associated with kidney diseases were significantly enriched in the proximal tubule- (PT) and injured PT-specific open chromatin regions in samples from patients with DKD. BACH1 was identified as a core transcription factor of injured PT cells; its binding target genes were highly associated with fibrosis and inflammation, which were also key features of injured PT cells. Additionally, Hi-C analysis revealed global chromatin architectural changes in DKD, accompanied by changes in local open chromatin patterns. Combining the snATAC-seq and Hi-C data identified direct target genes of BACH1, and indicated that BACH1 binding regions showed increased chromatin contact frequency with promoters of their target genes in DKD. Thus, our multi-omics analysis revealed BACH1 target genes in injured PTs and highlighted the role of BACH1 as a novel regulator of tubular inflammation and fibrosis.

Flexible & Stretchable EMG Sensor for Lower Extremity Amputee.

EMG signals can be widely used for indicators of muscle activity, and it can be used for robot control. However, the practical use of the EMG sensor for the amputee has been limited due to harsh conditions in the socket where strong pressure and friction exist. In this paper, thus we suggested a flexible and stretchable EMG Sensor. It is designed to withstand the pressure of the socket and to be used repeatedly with soft adhesive material. The performance of mechanical and electrical properties is investigated, and the muscle signals are recorded in static and dynamic (jump and gait) conditions. The selectivity of the recorded muscle signals during dorsiflexion and plantar flexion shows better than that of commercial electrodes indicating that it could be used for control of robotic legs in the future.Clinical Relevance- The flexible material and stretchable electrode pattern could be helpful in clinical research for an amputee.

Hemoglobin Subunit Theta 1 Promotes Proliferation by Reducing Reactive Oxygen Species in Lung Adenocarcinoma.

Lung adenocarcinoma is a crucial contributor to cancer-related mortality; however, effective treatments remain challenging. The present study aimed to investigate the role of hemoglobin subunit theta 1 (HBQ1), an α subunit of hemoglobin whose expression has recently been reported in non-erythroid cells, in lung adenocarcinoma. Comparative analysis showed that HBQ1 expression was significantly higher in lung adenocarcinoma tissues compared to normal lung tissues. Moreover, high HBQ1 expression was correlated with unfavorable overall survival and progression-free survival in patients, highlighting its potential as a prognostic marker. Our functional experiments revealed that when overexpressed, HBQ1 acts as an oncogene, enhancing cell proliferation, whereas HBQ1 knockdown inhibits it. Additionally, HBQ1 exhibited antioxidant properties by reducing basal reactive oxygen species levels, playing a crucial role in lung adenocarcinoma progression. These findings emphasize the critical role of HBQ1 in driving tumor growth and progression in lung adenocarcinoma. Our in vivo studies further supported the role of HBQ1 in lung adenocarcinoma. HBQ1 knockdown resulted in the inhibition of lung adenocarcinoma growth, demonstrating the potential of HBQ1 as a therapeutic target. Our findings highlight the importance of HBQ1 in lung adenocarcinoma and suggest its potential as both a diagnostic marker and a molecular target for therapeutic interventions.

AI-Based Geometry Recognition for Evaluating the Feasibility of Intensified Reaction and Separation Systems.

Reactive distillation (RD) provides notable advantages over conventional processes, regarding reduced energy requirements and CO2 emissions. However, as the benefits of RD may not be universally applicable, a comprehensive feasibility assessment is necessary. This study introduced an automated feasibility evaluation procedure for an RD column using an AI-based region recognition approach, reducing the reliance on expert knowledge and heuristics in graphical methods. Through k-means clustering-based image segmentation, topological information on the reaction and separation reachable region was extracted from ternary diagram landscapes. Subsequently, the extracted information was integrated into tray-by-tray calculations to automate the evaluation. This geometric calculation procedure was applied to assess the feasibility of RD columns with different types of reactions. The feasibility results were obtained within seconds, demonstrating the efficiency of the proposed approach. Furthermore, case studies validated the feasibility of the evaluation results for three practical examples using rigorous simulations, confirming its reliability and applicability.

A bivalent form of a RBD-specific synthetic antibody effectively neutralizes SARS-CoV-2 variants.

Coronavirus Disease 2019 (COVID-19) pandemic is severely impacting the world, and tremendous efforts have been made to deal with it. Despite many advances in vaccines and therapeutics, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants remains an intractable challenge. We present a bivalent Receptor Binding Domain (RBD)-specific synthetic antibody, specific for the RBD of wild-type (lineage A), developed from a non-antibody protein scaffold composed of LRR (Leucine-rich repeat) modules through phage display. We further reinforced the unique feature of the synthetic antibody by constructing a tandem dimeric form. The resulting bivalent form showed a broader neutralizing activity against the variants. The in vivo neutralizing efficacy of the bivalent synthetic antibody was confirmed using a human ACE2-expressing mouse model that significantly alleviated viral titer and lung infection. The present approach can be used to develop a synthetic antibody showing a broader neutralizing activity against a multitude of SARS-CoV-2 variants.

Engineering of a Fluorescent Protein for a Sensing of an Intrinsically Disordered Protein through Transition in the Chromophore State.

Intrinsically disordered proteins (IDPs) not only play important roles in biological processes but are also linked with the pathogenesis of various human diseases. Specific and reliable sensing of IDPs is crucial for exploring their roles but remains elusive due to structural plasticity. Here, we present the development of a new type of fluorescent protein for the ratiometric sensing and tracking of an IDP. A ß-strand of green fluorescent protein (GFP) was truncated, and the resulting GFP was further engineered to undergo the transition in the absorption maximum upon binding of a target motif within amyloid-ß (Aß) as a model IDP through rational design and directed evolution. Spectroscopic and structural analyses of the engineered truncated GFP demonstrated that a shift in the absorption maximum is driven by the change in the chromophore state from an anionic (460 nm) state into a neutral (390 nm) state as the Aß binds, allowing a ratiometric detection of Aß. The utility of the developed GFP was shown by the efficient and specific detection of an Aß and the tracking of its conformational change and localization in astrocytes.

3D-Printed Thermoplastic Polyurethane Electrodes for Customizable, Flexible Lithium-Ion Batteries with an Ultra-Long Lifetime.

3D printing technology has demonstrated great potential in fabricating flexible and customizable high-performance batteries, which are highly desired in the forthcoming intelligent and ubiquitous energy era. However, a significant performance gap, especially in cycling stability, still exists between the 3D-printed and conventional electrodes, seriously limiting the practical applications of 3D-printed batteries. Here, for the first time, a series of thermoplastic polyurethane (TPU)-based 3D-printed electrodes is developed via fused deposition modeling for flexible and customizable high-performance lithium-ion batteries. The TPU-based electrode filaments in kilogram order are prepared via a facile extrusion method. As a result, the electrodes are well-printed with high dimensional accuracy, flexibility, and mechanical stability. Notably, 3D-printed TPU-LFP electrodes exhibit a capacity retention of 100% after 300 cycles at 1C, which is among the best cycling performance of all the reported 3D-printed electrodes. Such excellent performance is associated with the superb stress cushioning properties of the TPU-based electrodes that can accommodate the volume change during the cycling and thus significantly prevent the collapse of 3D-printed electrode structures. The findings not only provide a new avenue to achieve customizable and flexible batteries but also guide a promising way to erase the performance gap between 3D-printed and conventional lithium-ion batteries.

Hybrid Artificial Solid Electrolyte Interphase with Dendrite-Free Lithium Deposition and High Ion Transport Kinetics.

Interfacial issues and dendritic Li deposition in lithium metal batteries (LMBs) hamper the practical application of liquid or solid-state cells. Here, a hybrid solid electrolyte interphase (SEI), based on hydroxyl-functionalized boron nitride (BN) nanosheets and poly(vinyl alcohol), is designed to solve the unstable nature of the Li anode-electrolyte interface. Rather than acquiring a rich Li halide environment through intense electrolyte decomposition, the hybrid SEI effectively regulates electrolyte decomposition and guarantees uniform Li plating via boosting interfacial Li+ ion transport at the interface. The Li+ ion boosting kinetics were deeply analyzed using simulations and spectroscopic analysis. It is revealed that the hydroxyl-functionalized BN can decrease kinetic energy barriers for Li+ ions and strongly holds TFSI- ions, thereby ensuring faster Li+ ion migration between electrodes and electrolytes. Tailoring the interfacial Li+ ion dynamics with hybrid SEI renders the Li transference number enhancement from 0.391 to 0.562 and 0.178 to 0.327 in liquid and solid-state cells, respectively. Moreover, Li symmetric cells with hybrid SEI exhibit an ultrahigh stability over 3500 h at 2 mA cm-2 with 2 mA h cm-2, along with the improved solid-state LMB performances. Our results suggest increasing Li+ ion transport at the interface is an alternative to resolve Li anode issues.

Genome wide association study to detect genetic regions related to isoflavone content in a mutant soybean population derived from radiation breeding.

Isoflavones are major secondary metabolites that are exclusively produced by legumes, including soybean. Soy isoflavones play important roles in human health as well as in the plant defense system. The isoflavone content is influenced by minor-effect quantitative trait loci, which interact with polygenetic and environmental factors. It has been difficult to clarify the regulation of isoflavone biosynthesis because of its complex heritability and the influence of external factors. Here, using a genotype-by-sequencing-based genome-wide association mapping study, 189 mutant soybean genotypes (the mutant diversity pool, MDP) were genotyped on the basis of 25,646 high-quality single nucleotide polymorphisms (SNPs) with minor allele frequency of >0.01 except for missing data. All the accessions were phenotyped by determining the contents of 12 isoflavones in the soybean seeds in two consecutive years (2020 and 2021). Then, quantitative trait nucleotides (QTNs) related to isoflavone contents were identified and validated using multi-locus GWAS models. A total of 112 and 46 QTNs related to isoflavone contents were detected by multiple MLM-based models in 2020 and 2021, respectively. Of these, 12 and 5 QTNs were related to more than two types of isoflavones in 2020 and 2021, respectively. Forty-four QTNs were detected within the 441-Kb physical interval surrounding Gm05:38940662. Of them, four QTNs (Gm05:38936166, Gm05:38936167, Gm05:38940662, and Gm05:38940717) were located at Glyma.05g206900 and Glyma.05g207000, which encode glutathione S-transferase THETA 1 (GmGSTT1), as determined from previous quantitative trait loci annotations and the literature. We detected substantial differences in the transcript levels of GmGSTT1 and two other core genes (IFS1 and IFS2) in the isoflavone biosynthetic pathway between the original cultivar and its mutant. The results of this study provide new information about the factors affecting isoflavone contents in soybean seeds and will be useful for breeding soybean lines with high and stable concentrations of isoflavones.

Identification of Loci Governing Agronomic Traits and Mutation Hotspots via a GBS-Based Genome-Wide Association Study in a Soybean Mutant Diversity Pool.

In this study, we performed a genotyping-by-sequencing analysis and a genome-wide association study of a soybean mutant diversity pool previously constructed by gamma irradiation. A GWAS was conducted to detect significant associations between 37,249 SNPs, 11 agronomic traits, and 6 phytochemical traits. In the merged data set, 66 SNPs on 13 chromosomes were highly associated (FDR p < 0.05) with the following 4 agronomic traits: days of flowering (33 SNPs), flower color (16 SNPs), node number (6 SNPs), and seed coat color (11 SNPs). These results are consistent with the findings of earlier studies on other genetic features (e.g., natural accessions and recombinant inbred lines). Therefore, our observations suggest that the genomic changes in the mutants generated by gamma irradiation occurred at the same loci as the mutations in the natural soybean population. These findings are indicative of the existence of mutation hotspots, or the acceleration of genome evolution in response to high doses of radiation. Moreover, this study demonstrated that the integration of GBS and GWAS to investigate a mutant population derived from gamma irradiation is suitable for dissecting the molecular basis of complex traits in soybeans.

Enhanced half-life and antitumor activity of interleukin-15 through genetic fusion of a serum albumin-specific protein binder.

Human interleukin-15 (hIL-15) has attracted a considerable attention as a promising cancer immunotherapeutic due to its function to directly stimulate the proliferation and cytotoxic activity of NK and T cells. Nevertheless, a relatively short half-life of hIL-15 requires repeated administration and higher doses, causing serious side effects. Here, we demonstrate an enhanced blood half-life and biological activity of hIL-15 through genetic fusion of a human serum albumin-specific protein binder (rHSA). The fusion construct (rHSA-IL15) was observed to maintain respective binding activities for both hIL-15 receptor α and human serum albumin. The rHSA-IL15 led to a significant increase in the secretion of Granzyme B and INF-γ by immune cells compare to free hIL-15, expanding the population of activated T cell subset such as CD4 + T and CD8+ T cells. The terminal half-life of the rHSA-IL15 was prolonged by around a 40-fold in transgenic mice expressing human serum albumin, compared to free hIL-15. The rHSA-IL15 resulted in distinct anti-tumor activities in xenograft SCC (squamous cell carcinoma) mouse and allograft melanoma mouse models through activation of NK and CD8+ T cells. The rHSA-IL15 is expected to be used in cancer immunotherapy, assisting in the development of other cytokines as immunotherapeutic agents with greater efficacy.

Ecological Responses of Nannophya koreana (Odonata: Libellulidae) to Temperature: Following Converse Bergmann's Rule.

Ecological rules such as Bergmann's rule and the temperature-size rule state that body-size decline is a universal response to warm temperatures in both homeotherms and poikilotherms. In the present study, we investigated the biological responses of Nannophya koreana, an endangered dragonfly species in Korea, by comparing body size in two habitats with large differences in water temperature, Mungyong-si (MG, terraced paddy fields) and Muui-do (MU, a mountainous wetland). To conserve the dragonfly populations, the collected larvae were photographed and released, and their head widths and body lengths were measured. There was no difference in the annual mean air temperature and precipitation between the two sites; however, the annual mean water temperature was substantially lower in MU than in MG. There was little difference in larval head width between the two sites; however, body length in the MU population was smaller than that in the MG population. Larval growth rate per 100-degree-days was 0.75 mm for MG and 1.16 for MU. The relationship between temperature and body size of N. koreana larvae showed opposite trends to Bergmann's rule and the temperature-size rule. Since the larval growth period during a year in MU was shorter than that in MG, the MU population potentially exhibits a higher growth rate as a mechanism of compensating for the low water temperature. Our study established the relationship between temperature and body size of N. koreana in two wetlands that had an obvious difference in water temperature despite being geographically close. The results highlight the importance of considering detailed factors such as habitat type when studying the temperature-size responses of organisms.

Computational epitope binning reveals functional equivalence of sequence-divergent paratopes.

The therapeutic efficacy of a protein binder largely depends on two factors: its binding site and its binding affinity. Advances in in vitro library display screening and next-generation sequencing have enabled accelerated development of strong binders, yet identifying their binding sites still remains a major challenge. The differentiation, or "binning", of binders into different groups that recognize distinct binding sites on their target is a promising approach that facilitates high-throughput screening of binders that may show different biological activity. Here we study the extent to which the information contained in the amino acid sequences comprising a set of target-specific binders can be leveraged to bin them, inferring functional equivalence of their binding regions, or paratopes, based directly on comparison of the sequences, their modeled structures, or their modeled interactions. Using a leucine-rich repeat binding scaffold known as a "repebody" as the source of diversity in recognition against interleukin-6 (IL-6), we show that the "Epibin" approach introduced here effectively utilized structural modelling and docking to extract specificity information encoded in the repebody amino acid sequences and thereby successfully recapitulate IL-6 binding competition observed in immunoassays. Furthermore, our computational binning provided a basis for designing in vitro mutagenesis experiments to pinpoint specificity-determining residues. Finally, we demonstrate that the Epibin approach can extend to antibodies, retrospectively comparing its predictions to results from antigen-specific antibody competition studies. The study thus demonstrates the utility of modeling structure and binding from the amino acid sequences of different binders against the same target, and paves the way for larger-scale binning and analysis of entire repertoires.

Mutation effects on FAS1 domain 4 based on structure and solubility.

Mutations in the fasciclin 1 domain 4 (FAS1-4) of transforming growth factor ß-induced protein (TGFBIp) are associated with insoluble extracellular deposits and corneal dystrophies (CDs). The decrease in solubility upon mutation has been implicated in CD; however, the exact molecular mechanisms are not well understood. Here, we performed molecular dynamics simulations followed by solvation thermodynamic analyses of the FAS1-4 domain and its three mutants-R555W, R555Q, and A546T-linked to granular corneal dystrophy type 1, Thiel-Behnke corneal dystrophy and lattice corneal dystrophy, respectively. We found that both R555W and R555Q mutants have less affinity toward solvent water relative to the wild-type protein. In the R555W mutant, a remarkable increase in solvation free energy was observed because of the structural changes near the mutation site. The mutation site W555 is buried in other hydrophobic residues, and R557 simultaneously forms salt bridges with E554 and D561. In the R555Q mutant, the increase in solvation free energy is caused by structural rearrangements far from the mutation site. R558 separately forms salt bridges with D575, E576, and E598. Thus, we thus identified the relationship between the decrease in solubility and conformational changes caused by mutations, which may be useful in designing potential therapeutics and in blocking FAS1 aggregation related to CD.

Analysis of genetic diversity and relationships of Perilla frutescens using novel EST-SSR markers derived from transcriptome between wild-type and mutant Perilla.

BACKGROUND: Perilla frutescens (Lamiaceae) is distributed in East Asia and is classified into var. frutescens and crispa. P. frutescens is multipurpose crop for human health because of a variety of secondary metabolites such as phenolic compound and essential oil. However, a lack of genetic information has hindered the development and utilization of Perilla genotypes. METHODS AND RESULTS: This study was performed to develop expressed sequence tag-simple sequence repeat (EST-SSR) markers from P. frutescens var. crispa (wild type) and Antisperill (a mutant cultivar) and used them to assess the genetic diversity of, and relationships among, 94 P. frutescens genotypes. We obtained 65 Gb of sequence data comprising 632,970 transcripts by de novo RNA-sequencing. Of the 14,780 common SSRs, 102 polymorphic EST-SSRs were selected using in silico polymerase chain reaction (PCR). Overall, successful amplification from 58 EST-SSRs markers revealed remarkable genetic diversity and relationships among 94 P. frutescens genotypes. In total, 268 alleles were identified, with an average of 4.62 alleles per locus (range 2-11 alleles/locus). The average polymorphism information content (PIC) value was 0.50 (range 0.04-0.86). In phylogenetic and population structure analyses, the genotypes formed two major groups: Group I (var. crispa) and Group II (var. frutescens). CONCLUSION: This results suggest that 58 novel EST-SSR markers derived from wild-type cultivar (var. crispa) and its mutant cultivar (Antisperill) have potential uses for population genetics and recombinant inbred line mapping analyses, which will provide comprehensive insights into the genetic diversity and relationship of P. frutescens.