Peanut (Arachis hypogaea), a globally important oilseed crop, exhibits contrasting growth habits between wild species (prostrate) and cultivated varieties (erect or spreading), but the underlying mechanism is unclear. In this study, we performed quantitative trait locus (QTL) mapping of two recombinant inbred line (RIL) populations of peanut using SNP arrays and bulk segregant analysis, which identified qGH15 on chromosome 15 as a major QTL regulating growth habit. Fine mapping using KASP markers narrowed the candidate region to a 151-kb interval, while analysis of a residual heterozygous line (RHL) further delimited qGH15 to a 38-kb interval containing a single candidate gene, which we designated as AhGH15. Genotyping of a natural population revealed multiple types of polymorphisms in this gene associated with the erect habit in cultivated varieties. Phylogenetic and pedigree analyses demonstrated that these polymorphisms were recurrently and convergently selected during peanut domestication and breeding, with modern hybridization accelerating their dissemination. Transcriptome deep sequencing and gene co-expression analysis via WGCNA revealed polymorphism-specific patterns of transcriptome regulation, with co-expressed gene modules differentially active between accessions with erect or prostrate growth habits. These findings establish AhGH15 as a key determinant of peanut growth habit and highlight its complex selection history during peanut improvement.

Asparagi Radix (Asparagus cochinchinensis) is a commonly used Traditional Chinese Medicine and functional food, but it has been subjected to various types of adulteration because of the highly similar morphological characters among related species. To address this, we sequenced the complete chloroplast genomes of A. cochinchinensis and five reported adulterant species (A. filicinus, A. lycopodineus, A. subscandens, A. taliensis, and A. meioclados), among the chloroplast genome of A. subscandens was sequenced and assembled for the first time. Comparative analysis of the chloroplast genomes revealed some highly variable hotspot regions suitable for DNA mini-barcode authentication. Using high-resolution melting (HRM) analysis, we developed species-specific melting profiles from two selected mini-barcodes (i.e., trnD-trnY and rbcL-accD), which clearly distinguished genuine Asparagi Radix from adulterants. These mini-barcodes were applied to test nine commercial products. Eight samples matched the melting profiles and temperature of A. cochinchinensis, while one was identified as adulterated. Further, ITS2 sequencing revealed that this adulterant exhibited superimposed chromatograms, and cloning-based sequencing confirmed the presence of two distinct ITS2 sequences with high similarity to A. cochinchinensis and A. taliensis, suggesting a possible hybrid origin. This study demonstrates that chloroplast genome-derived mini-barcodes combined with HRM provide an efficient and reliable method for authenticating Asparagi Radix.

Asparagi radix (Asparagus cochinchinensis) is a commonly used traditional Chinese medicine and functional food. It has been subjected to various types of adulteration because of the high similarity of morphological characters among its related species. We sequenced the complete chloroplast genomes of A. cochinchinensis and five reported adulterant species (Asparagus filicinus, Asparagus lycopodineus, Asparagus subscandens, Asparagus taliensis, and Asparagus meioclados) to address the above issue. Notably, the chloroplast genome of A. subscandens was sequenced and assembled for the first time. The comparative analysis of the chloroplast genomes revealed several highly variable hotspot regions suitable for DNA mini-barcode authentication. By using high-resolution melting (HRM) analysis, we developed species-specific melting profiles from two selected mini-barcodes (i.e., trnD–trnY and rbcL–accD), which clearly distinguished genuine Asparagi radix from adulterants. These mini-barcodes were applied to test nine commercial products. The melting profiles and temperatures of eight samples matched those of A. cochinchinensis, whereas one sample was identified as adulterated. Furthermore, ITS2 sequencing revealed that this adulterant exhibited superimposed chromatograms, and cloning-based sequencing confirmed the presence of two distinct ITS2 sequences with high similarity to A. cochinchinensis and A. taliensis, suggesting a possible hybrid origin. Our study demonstrates that chloroplast genome–derived mini-barcodes combined with HRM analysis provide an efficient and reliable method for authenticating Asparagi radix.

Peanut (Arachis hypogaea L.) is an important oil and edible protein crop. Its fatty acid composition not only influences the quality of peanut oil but also impacts flavor, shelf life, and consumer health. Peanut oil is comprised of approximately 80% oleic acid (C18:1) and linoleic acid (C18:2), 10% palmitic acid (C16:0), and the remaining 10% includes stearic acid (C18:0), arachidic acid (C20:0), gadoleic acid (C20:1), behenic acid (C22:0), and lignoceric acid (C24:0). To unravel the genetic foundation of fatty acid content and delve into QTL localization, high-density SNP microarrays were used to genotype the RIL population of ‘SunOleic 97R’ × ‘NC94022’. A genetic linkage map was constructed with 3,141 SNP markers, covering a total genetic distance of 3,051.81 cM. Sixty quantitative trait loci (QTLs) associated with fatty acids were distributed in 11 linkage groups, with phenotypic variance explained (PVE) ranging from 1.37 to 44.92%. Notably, the QTLs qFAT_A05.1 and qFAT_A08.1 are multiple-effect loci contributing to various fatty acid compositions. Moreover, 15 haplotypes for the QTLs qFAT_A05.1 and qFAT_A08.1 were identified through genotyping 178 peanut germplasms. Haplotype analysis in a natural population confirmed the close relationship of the QTLs with the contents of oil, oleic acid, lignoceric acid, palmitic acid and behenic acid. This study serves as a valuable reference for selecting improved peanut genotypes with superior oil quality and desirable fatty acid composition.

BACKGROUND:Peanut (Arachis hypogaea L.) is a vital global crop, frequently threatened by both abiotic and biotic stresses. Among the most damaging biotic stresses is Tomato spotted wilt virus (TSWV), which causes peanut spotted wilt disease resulting in significant yield loss. Developing TSWV-resistant cultivars is crucial to new cultivar release. Previous studies have used a subset of the "S" recombinant inbred line (RIL) population derived from SunOleic 97R and NC94022 and identified quantitative trait loci (QTLs) for resistance to TSWV. These studies utilized different genotyping techniques and found large consistent genomic regions on chromosome A01. The objective of this study was to fine map the QTL and identify candidate genes using the entire population of 352 RILs and high-density, high-quality peanut SNP arrays. RESULTS:We used both versions of the peanut SNP arrays with five years of disease ratings, and successfully mapped the long-sought peanut spotted wilt disease resistance locus, PSWDR-1. QTL analyses identified two major QTLs, explaining 41.43% and 43.69% of the phenotypic variance within 3.6 cM and 0.28 cM intervals using the peanut Axiom_Arachis-v1 and Axiom_Arachis-v2 SNP arrays, respectively, on chromosome A01. These QTLs corresponded to 295 kb and 235 kb physical intervals. The unique overlap region of these two QTLs was 488 kb. A comparison of the genetic linkage map with the reference genome revealed a 1.3 Mb recombination "cold spot" (11.325-12.646 Mb) with only two recombination events of RIL-S1 and RIL-S17, which displayed contrasting phenotypes. Sequencing of these two recombinants confirmed the cold spot with only five SNPs detected within this region. CONCLUSIONS:This study successfully identified a peanut spotted wilt disease resistance locus, PSWDR-1, on chromosome A01 within a recombination "cold spot". The PSWDR-1 locus contains three candidate genes, a TIR-NBS-LRR gene (Arahy.1PK53M), a glutamate receptor-like gene (Arahy.RI1BYW), and an MLO-like protein (Arahy.FX71XI). These findings provide a foundation for future functional studies to validate the roles of these candidate genes in resistance and application in breeding TSWV-resistant peanut cultivars.

Multiparent advanced generation intercross (MAGIC) populations are a new genetic resource for high-resolution mapping of quantitative traits and as a source of new germplasm or improved cultivars for breeding due to the high level of recombination events in the population. Here, we have developed an eight-founder MAGIC population for peanut (Arachis hypogaea L.) (PeanutMAGIC). Eight diverse founders were intercrossed using a simple funnel mating design to ensure that the MAGIC population would possess equal representation from each founder. This was followed by advancement using small family plot and single-seed descent, resulting in 3187 F2:7 recombinant inbred lines (RILs). The objective of this study was to introduce this PeanutMAGIC as a new resource for genetic and genomic studies. We randomly selected a smaller subset of 310 RILs (MAGIC Core) from PeanutMAGIC and conducted genotyping using whole genome sequencing and phenotyping over two growing seasons for seed and pod traits. Whole genome characterization of the MAGIC Core demonstrated that PeanutMAGIC harbors a balanced and evenly differentiated mosaic of genomic blocks from eight founders, providing unique recombination events for high-resolution mapping of quantitative traits. Using 2-year phenotypic data, we showed that PeanutMAGIC can improve genetic mapping power of a spectrum of qualitative, like seed coat color, to quantitative traits such as pod weight, seed weight, shelling percentage, pod constriction, and pod reticulation. These findings show that the PeanutMAGIC population can be used by the peanut research community as a new resource for genetic and genomic studies and for cultivar improvement.

Cadmium (Cd) is a hazardous heavy metal that threatens human health through the consumption of contaminated rice. To mitigate Cd accumulation in rice grains, it is crucial to reduce Cd uptake. Nevertheless, the transcriptional mechanisms governing Cd uptake in rice remain largely unknown. This research identifies the transcription factor OsNAC5 in Oryza sativa as a positive regulator of the Cd transporter gene OsNRAMP1, thereby influencing Cd uptake. OsNAC5 is predominantly expressed in the roots, resides in the nucleus, and is upregulated by Cd-induced hydrogen peroxide (H2O2). Knocking out OsNAC5 results in lower Cd concentrations in both shoots and roots and heightens sensitivity to Cd. The expression of OsNRAMP1, enhanced by Cd stress, is dependent on OsNAC5. OsNAC5 binds to "CATGTG" motifs in the OsNRAMP1 promoter, activating its expression. The loss of OsNAC5 function leads to reduced Cd accumulation in rice grains. Our findings provide insights into the transcriptional regulation of Cd stress response in rice and propose biotechnological strategies to lower Cd uptake in crops.

Abstract Background Aspergillus flavus is an important agricultural and food safety threat due to its production of carcinogenic aflatoxins. It has high level of genetic diversity that is adapted to various environments. Recently, we reported two reference genomes of A. flavus isolates, AF13 (MAT1-2 and highly aflatoxigenic isolate) and NRRL3357 (MAT1-1 and moderate aflatoxin producer). Where, an insertion of 310 kb in AF13 included an aflatoxin producing gene bZIP transcription factor, named atfC. Observations of significant genomic variants between these isolates of contrasting phenotypes prompted an investigation into variation among other agricultural isolates of A. flavus with the goal of discovering novel genes potentially associated with aflatoxin production regulation. Present study was designed with three main objectives: (1) collection of large number of A. flavus isolates from diverse sources including maize plants and field soils; (2) whole genome sequencing of collected isolates and development of a pangenome; and (3) pangenome-wide association study (Pan-GWAS) to identify novel secondary metabolite cluster genes. Results Pangenome analysis of 346 A. flavus isolates identified a total of 17,855 unique orthologous gene clusters, with mere 41% (7,315) core genes and 59% (10,540) accessory genes indicating accumulation of high genomic diversity during domestication. 5,994 orthologous gene clusters in accessory genome not annotated in either the A. flavus AF13 or NRRL3357 reference genomes. Pan-genome wide association analysis of the genomic variations identified 391 significant associated pan-genes associated with aflatoxin production. Interestingly, most of the significantly associated pan-genes (94%; 369 associations) belonged to accessory genome indicating that genome expansion has resulted in the incorporation of new genes associated with aflatoxin and other secondary metabolites. Conclusion In summary, this study provides complete pangenome framework for the species of Aspergillus flavus along with associated genes for pathogen survival and aflatoxin production. The large accessory genome indicated large genome diversity in the species A. flavus, however AflaPan is a closed pangenome represents optimum diversity of species A. flavus. Most importantly, the newly identified aflatoxin producing gene clusters will be a new source for seeking aflatoxin mitigation strategies and needs new attention in research.

Identification of candidate genes and molecular markers for late leaf spot (LLS) disease resistance in peanut (Arachis hypogaea) has been a focus of molecular breeding for the U.S. industry-funded peanut genome project. Efforts have been hindered by limited mapping resolution due to low levels of genetic recombination and marker density available in traditional biparental mapping populations. To address this, a multi-parental nested association mapping population has been genotyped with the peanut 58K single-nucleotide polymorphism (SNP) array and phenotyped for LLS severity in the field for 3 years. Joint linkage-based quantitative trait locus (QTL) mapping identified nine QTLs for LLS resistance with significant phenotypic variance explained up to 47.7%. A genome-wide association study identified 13 SNPs consistently associated with LLS resistance. Two genomic regions harboring the consistent QTLs and SNPs were identified from 1,336 to 1,520 kb (184 kb) on chromosome B02 and from 1,026.9 to 1,793.2 kb (767 kb) on chromosome B03, designated as peanut LLS resistance loci, PLLSR-1 and PLLSR-2, respectively. PLLSR-1 contains 10 nucleotide-binding site leucine-rich repeat disease resistance genes. A nucleotide-binding site leucine-rich repeat disease resistance gene, Arahy.VKVT6A, was also identified on homoeologous chromosome A02. PLLSR-2 contains five significant SNPs associated with five different genes encoding callose synthase, pollen defective in guidance protein, pentatricopeptide repeat, acyl-activating enzyme, and C2 GRAM domains-containing protein. This study highlights the power of multi-parent populations such as nested association mapping for genetic mapping and marker-trait association studies in peanuts. Validation of these two LLS resistance loci will be needed for marker-assisted breeding.

Light-harvesting chlorophyll a/b binding proteins (Lhcb) play crucial roles in plant growth, development, and the response to abiotic stress in higher plants. Previous studies have reported that Lhcb genes were involved in the phytochrome regulation and responded to different light and temperature conditions in Poaceae (such as maize). However, the evolution and functions of Lhcb genes remains poorly characterized in important Rosaceae species. In this investigation, we conducted a genome-wide analysis and identified a total of 212 Lhcb genes across nine Rosaceae species. Specifically, we found 23 Lhcb genes in Fragaria vesca, 20 in Prunus armeniaca, 33 in Malus domestica ‘Gala’, 21 in Prunus persica, 33 in Rosa chinensis, 29 in Pyrus bretschneideri, 18 in Rubus occidentalis, 20 in Prunus mume, and 15 in Prunus salicina. Phylogenetic analysis revealed that the Lhcb gene family could be classified into seven major subfamilies, with members of each subfamily sharing similar conserved motifs. And, the functions of each subfamily was predicted based on the previous reports from other species. The Lhcb proteins were highly conserved within their respective subfamilies, suggesting similar functions. Interestingly, we observed similar peaks in Ks values (0.1–0.2) for Lhcb genes in apple and pear, indicating a recent whole genome duplication event (about 30 to 45 million years ago). Additionally, a few Lhcb genes underwent tandem duplication and were located across all chromosomes of nine species of Rosaceae. Furthermore, the analysis of the cis-acting elements in the 2000 bp promoter region upstream of the pear Lhcb gene revealed four main categories: light response correlation, stress response correlation, hormone response correlation, and plant growth. Quantitative expression analysis demonstrated that Lhcb genes exhibited tissue-specific expression patterns and responded differently to low-temperature stress in Rosaceae species. These findings shed light on the evolution and phylogeny of Lhcb genes in Rosaceae and highlight the critical role of Lhcb in pear’s response to low temperatures. The results obtained provide valuable insights for further investigations into the functions of Lhcb genes in Rosaceae, and these functional genes will be used for further fruit tree breeding and improvement to cope with the current climate changes.

乡村旅游发展对实现生计可持续、促进城乡和区域共同富裕能够发挥建构性和工具性作用.选择2010-2020年中国30个省域的面板数据样本,采用双变量空间自相关检验和空间面板计量模型等研究方法,从整体与区域视角分析了中国乡村旅游发展促进农村可持续生计的空间溢出效应及其作用机制.研究表明:中国乡村旅游发展和农村可持续生计在时间上表现出高值省域的优势锁定,且均呈现由东南向西北递减的空间分异格局,在空间上二者之间具有显著的空间依赖与关联特征;乡村旅游发展能够通过内外部响应双重作用机制发挥其对传统生计替代效应的适应性,扩大增权效应、增强内源性发展动力实现本地区农村生计可持续,并通过空间溢出效应带动邻接地区农村生计可持续发展;乡村旅游发展在生计脆弱性、生计资本、生计恢复力和生计结果的多维要素作用下影响农村可持续生计,呈现出直接效应"由西部向中部和东部递减"、空间溢出效应"由中部向西部和东部递减"的区域异质性.

A rapid, simple, and sensitive fluorescent detection method for brown spot of tobacco is established by lambda exonuclease-induced Mg2+-dependent DNAzyme amplification. It contains hybridization of the Alternaria alternata genome and HP1, digestion of the 5'-phosphorylated strand of the hybrid dsDNA by lambda exonuclease, acquisition of complete Mg2+-dependent DNAzyme, cleavage of the substrate modified with FAM and BHQ-1, and fluorescent detection. The proposed assay exhibits good sensitivity (10 pg L-1), selectivity and reproducibility. The method does not require pure DNA and expensive instruments, and can be performed within 2.5 hours. To the best of our knowledge, this is the first report of fluorescent detection of Alternaria alternata and its tobacco field samples. This method can be applied to the rapid and sensitive detection of Alternaria alternata in tobacco and its seedlings, and is particularly important for the green prevention and control of tobacco brown spot disease.

为了明确河川沙塘鳢(Odontobutis potamophila Günther)养殖宜投喂饵料生物的种类和规格,通过室内外试验研究了沙塘鳢对生活习性不同8种饵料生物的种类选择,对不同规格赤眼鳟(Squaliobarbus curriculus Richardson)的选择,以及其不同生长阶段对饵料鱼大小的选择.研究结果表明,沙塘鳢对麦鲮(Cirrhinus mrigala Hamilton)的选择指数显著大于其对剩余7种饵料生物的选择指数;在无仿真水草条件下,沙塘鳢对小规格饵料生物的选择指数显著大于对大、中规格饵料生物的选择指数;在仿真水草条件下,沙塘鳢对小规格饵料生物的选择指数显著大于其对大规格饵料生物的选择指数.在不同生长阶段中,沙塘鳢全长与其捕食的饵料鱼全长呈现正相关关系;沙塘鳢捕食的饵料鱼PPR值为0.23—0.73,均值为0.49±0.1(均值±标准差);随沙塘鳢规格的增加其捕食饵料鱼的PPR值降低,捕食饵料鱼规格分布小于环境中饵料鱼规格分布.综上,沙塘鳢养殖宜投喂饵料鱼麦鲮,饵料鱼PPR值宜为0.38—0.6.

Introduction: Tirbanibulin is a novel inhibitor of tubulin polymerization developed as a topical formulation for AK that has shown to be safe and superior to vehicle in AK clearance at 2 months after 5-days once-daily self-application.

Vitamin A is essential for brain function, in addition to its important roles in vision, immunity, and reproduction. Previous studies have shown that retinoic acid (RA), the bioactive form of vitamin A, is involved in the regulation of various intracellular responses related to biological rhythms. RA is reported to affect the circadian rhythm by binding to RA receptors, such as receptors in the circadian feedback loops in the mammalian suprachiasmatic nucleus. However, evidence of the impacts of vitamin A deficiency (VAD) on biological rhythms is limited, and most of the related studies were conducted on animals. In this review, we described the physiological functions of biological rhythms and physiological pathways/molecular mechanisms regulating the biological rhythms. We then discussed the current understanding of the associations of VAD with biological rhythm disorders/diseases (sleep disorders, impairments in learning/memory, emotional disorders, and other immune or metabolism diseases) and summarized the currently proposed mechanisms (mainly by retinoid nuclear receptors and related proteins) for the associations. This review may help recognize the role of VAD in biological rhythm disorders and stimulate clinical or epidemiological studies to confirm the findings of related animal studies.

Background Protein post-translational modification (PTM) is a key issue to investigate the mechanism of protein’s function. With the rapid development of proteomics technology, a large amount of protein sequence data has been generated, which highlights the importance of the in-depth study and analysis of PTMs in proteins. Method We proposed a new multi-classification machine learning pipeline MultiLyGAN to identity seven types of lysine modified sites. Using eight different sequential and five structural construction methods, 1497 valid features were remained after the filtering by Pearson correlation coefficient. To solve the data imbalance problem, Conditional Generative Adversarial Network (CGAN) and Conditional Wasserstein Generative Adversarial Network (CWGAN), two influential deep generative methods were leveraged and compared to generate new samples for the types with fewer samples. Finally, random forest algorithm was utilized to predict seven categories. Results In the tenfold cross-validation, accuracy (Acc) and Matthews correlation coefficient (MCC) were 0.8589 and 0.8376, respectively. In the independent test, Acc and MCC were 0.8549 and 0.8330, respectively. The results indicated that CWGAN better solved the existing data imbalance and stabilized the training error. Alternatively, an accumulated feature importance analysis reported that CKSAAP, PWM and structural features were the three most important feature-encoding schemes. MultiLyGAN can be found at https://github.com/Lab-Xu/MultiLyGAN . Conclusions The CWGAN greatly improved the predictive performance in all experiments. Features derived from CKSAAP, PWM and structure schemes are the most informative and had the greatest contribution to the prediction of PTM.

Sludge foaming has been a persistent ailment in worldwide municipal sewage treatment plants (MSTPs). In particular, this problem frequently occurs and lasts throughout the winter and spring season in the sub-plateau MSTPs. Aims to explore the causes of sludge foaming under sub-plateau circumstance, the correlation between the structural and functional changes in bacterial communities and sewage properties was conducted using Illumina high-throughput sequencing technology. The results from 16S rRNA gene sequencing shown that the relative abundances of Actinobacteria (39.5 % +/- 4.8 %) and Firmicutes (10.5 % +/- 1.0 %) in the foaming sludge were remarkably higher than that in the non-foaming sludge. The abundances of Candidatus Microthrix parvicella (29.8 % +/- 6.9 %), Trichococcus (6.5 % +/- 1.0 %), and Ornithinibacter (3.5 % +/- 0.6 %) were increased after sludge foaming, and which are considered as the dominated foaming-causative bacterial assemblages by the linear discriminant analysis (LDA). Spearman's correlation analysis and redundancy analysis (RDA) also agreed that lower temperature and dissolved oxygen (DO) levels, higher lipid and protein contents were significantly correlated with sludge foaming. In addition, the predictive functional profiling indicated that the enzymes and their coding genes of the bacterial communities are mainly involved in the fast proliferation and lipid metabolism, and which abundance are highly upregulated during foaming period. These results suggested that the sludge foaming was strongly linked to the Actinobacteria and Firmicutes communities and higher lipid and protein content in the sewage under sub-plateau circumstance. The findings might be important for developing operating strategies to maintain the desired microbial community pattern and improve the performance of MSTPs.

Seed dormancy is an important breeding trait for the development of certain types of peanut cultivars. Peanut cultivars with seed dormancy can inhibit preharvest sprouting in which the sprouting may increase susceptibility to preharvest aflatoxin contamination. The recombinant inbred line (RIL) mapping population derived from a cross of Tifrunner, a dormant Runner type, and GT-C20, a non-dormant Spanish type, were planted in the field for 2 years, and the freshly harvested seeds were used for seed dormancy tests at 7, 14, 21, and 28 days during germination. There were three RILs from 2-year tests with no dormancy (T48, T83, T160) and two lines with strong dormancy (T11, T163). This RIL population was genotyped using peanut SNP array ‘Axiom_Arachis’ 58 K, and two major seed dormancy QTLs were anchored on chromosome A04 and A05 with 43.16% and 51.61% of the phenotype variation explained (PVE), respectively. The QTL mapped on chromosome A05 had been anchored on a physical map interval of 98 kb (157.538–157.636 Mb) from which a possible candidate gene (Arahy.KB746A, ethylene-responsive transcription factor) was identified. Reference to the peanut physical map and flanking sequences, DNA markers can be developed for these two QTLs and used in marker-assisted breeding selection for seed dormancy in peanut.

采用静态急性毒性试验研究了3种常用水产用药物对河川沙塘鳢(Odontobutis potamophila)幼鱼24、48、72 h半数致死质量浓度ρ(LC50)(95％置信区间)以及安全质量浓度(SC).结果表明,10％聚维酮碘对河川沙塘鳢幼鱼24、48、72 h的半数致死质量浓度ρ分别为1466、1148、997.6 mg/L,安全浓度为270.0 mg/L;40％辛硫磷对河川沙塘鳢幼鱼24、48、72 h的半数致死质量浓度ρ分别为4.030、1.207、0.409 mg/L,安全浓度为0.1085 mg/L;4.5％高效氯氰菊酯对河川沙塘鳢幼鱼24、48、72 h的半数致死质量浓度ρ分别为3.007、2.695、0.9740 mg/L,安全浓度为0.7246 mg/L.