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Leonurus japonicus Houtt. ( L. japonicus ), as an important plant resource with both ornamental and medicinal value, has now spread worldwide and is widely studied. Currently, its chromosomal genome and chloroplast genome (cpDNA) have been reported, but the mitochondrial genome (mtDNA) has not yet been explored. In this study, we extracted DNA from fresh leaves of L. japonicus and performed sequencing and assembly of its mtDNA using both second-generation and third-generation sequencing technologies. The complete mtDNA of L. japonicus is 382,905 bp in length, with a GC content of 45.13%. This genome includes 15 tRNA genes, 32 protein-coding genes (PCGs), and 4 rRNA genes. In this mtDNA genome, we predicted a total of 480 RNA editing sites among the 32 PCGs. Subsequently, we conducted analyses on repetitive sequences, organelle genome sequence migration, and Relative Synonymous Codon Usage (RSCU). There are 28 homologous sequence fragments between the mtDNA and cpDNA of L. japonicus , which are related to the migration of 10 mtDNA genes. The RSCU analysis predicted 28 high-frequency codons, most of which prefer to end with A/U. Selection pressure analysis indicated that the Ka/Ks ratio for the majority of PCGs is less than 1, suggesting they are highly conserved during evolutionary processes. Phylogenetic results from 24 species indicate that the genera Leonurus and Scutellaria within the Lamiaceae family have the closest relationships. In summary, we have successfully assembled the complete mtDNA of L. japonicus by integrating second-generation and third-generation sequencing data for the first time. Subsequent multi-faceted analyses have allowed us to gain deeper insights into the numerous features of this genome, providing important reference data for the molecular genetics, dynamic evolution, and species identification of this plant. This work promotes the conservation and development of this important resource of medicinal and edible plants.

Background Leonurus japonicus ( L. japonicus ) is a herbaceous flowering plant, widely distributed in Asia. Drought is one of the primary environmental stress factors affecting L. japonicus growth. Previous studies have demonstrated that WRKY transcription factors (TFs) play a crucial role in plant responses to drought stress. So far, there has been no research on the function of WRKY genes in L. japonicus . Results The physiological experiment results showed that drought stress significantly increased the malondialdehyde (MDA), proline, and hydrogen peroxide (H₂O₂) content of L. japonicus . Transcriptome analysis revealed significant changes in the expression levels of the WRKY gene family. Based on bioinformatics analysis, 67 WRKY genes ( LjWRKYs ) were identified in the genome of L. japonicus , with amino acid lengths ranging from 85 to 574. The LjWRKYs can be divided into three subfamilies. Among them, the expression of LjWRKY (1/4/23/44) were significantly up-regulated under drought stress, whereas the expression of LjWRKY (21/25/65) were significantly down-regulated. Additionally, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that after drought stress, differentially expressed genes (DEGs) were enriched in plant hormone signal transduction pathway, the MAPK signaling pathway and biosynthesis of secondary metabolites pathway. In the MAPK pathway, there were 19 DEGs, 9 of which contained W-box regions, suggesting that they may be potential regulatory targets of LjWRKY TFs under drought stress. Conclusion These findings suggested that WRKY gene family may participate in the response to drought stress in L. japonicus . This study provides a scientific basis for the further development and functional validation of the WRKY gene family in L. japonicus. Clinical trial number Not applicable.

Chinese motherwort ( Leonurus japonicus ), a member of Lamiaceae family, is a commonly used medicinal herb for treating obstetrical and gynecological diseases, producing over 280 officinal natural products. Due to limited genomic resources, little progress has been made in deciphering the biosynthetic pathway of valuable natural products in L. japonicus . Here, we de novo assembled the L. japonicus genome using high-coverage ONT long reads and Hi-C reads. The chromosome-level genome assembly contained ten chromosomes representing 99.29% of 489.34 Mb genomic sequence with a contig and scaffold N50 of 7.27 Mb and 50.86 Mb, respectively. Genome validations revealed BUSCO and LAI score of 99.2% and 21.99, respectively, suggesting high quality of genome assembly. Using transcriptomic data from various tissues, 22,531 protein-coding genes were annotated. Phylogenomic analysis of 13 angiosperm plants suggested L. japonicus had 58 expanded gene families functionally enriched in specialized metabolism such as diterpenoid biosynthesis. The genome assembly, annotation, and sequencing data provide resources for the elucidation of biosynthetic pathways behind natural products of pharmaceutical applications in L. japonicus .

This study examines the production of five phenolic acids (chlorogenic acid, neochlorogenic acid, ferulic acid, caffeic acid and p-coumaric acid) following over-expression of AtPAP1 transcription factor by four transgenic root clones of Leonurus sibiricus after Agrobacterium rhizogenes transformation. The AtPAP1 expression level was estimated by quantitative real-time PCR. High levels of phenolic acids were found in the transgenic roots of L. sibiricus and were determined by high-performance liquid chromatography–mass spectrometry analysis. Additionally, transgenic roots showed antimicrobial potential and cytotoxic activity on glioma cells in IV grade. Our results suggest that L. sibiricus transformed roots with AtPAP1 gene over-expression may represent a potential source of phenolic acids.

Leonurus cardiaca is well known for its medicinal importance. In this investigation, genotypic characterization of this species from six eco-geographical regions of Iran was evaluated by four molecular techniques (AFLP, RAPD, ISSR and IRAP). A total of 899 polymorphic fragments were detected by used molecular markers (AFLP = 356, RAPD = 325, ISSR = 113 and IRAP = 105) with an overall average polymorphism of 81.24 %. Genetic variation calculated using Shannon’s Information index (I) and Nei’s gene diversity index (H) showed high genetic diversity in studied germplasm. Also, analysis of molecular variance showed high genetic variation among (55 %) and within populations (45 %). UPGMA dendrogram constructed from combined data of molecular markers distinguished studied populations in accordance with the results obtained by each marker which all individuals were clearly differentiated into two major clusters. The correlation coefficients were statistically significant for all marker systems with the highest correlation between similarity matrixes of RAPD and ISSR markers (r = 0.82). The present results have an important implication for L. cardiaca germplasm characterization, improvement, and conservation. Furthermore, the characterized individuals exhibited a great deal of molecular variation and they seem to have a rich gene pool for breeding programs.

LJAMP1 is a small antimicrobial protein purified previously from the seeds of motherwort, and it is expressed preferentially in seeds. A 794-bp upstream sequence of the ATG start codon was isolated using a genome walking method and cloned into the upstream of the β-glucuronidase (GUS) reporter gene to determine the GUS tissue-specific expression pattern. The transgenic tobacco showed that pLJAMP1 promoter derived GUS reporter gene special expression in pollen, achene and seed. The analysis of cis-acting elements also revealed pLJAMP1 promoter contained pollen and seed related transcriptional control elements.

Background Leonurus japonicus , a significant medicinal plant known for its therapeutic effects on gynecological and cardiovascular diseases, has genetic diversity that forms the basis for germplasm preservation and utilization in medicine. Despite its economic value, limited research has focused on its genetic diversity and divergence. Results The avg. nucleotide diversity of 59 accessions from China were 0.00029 and hotspot regions in petN-psbM and rpl32-trnL (UAG) spacers, which can be used for genotype discrimination. These accessions divided into four clades with significant divergence. The four subclades, which split at approximately 7.36 Ma, were likely influenced by the Hengduan Mountains uplift and global temperature drop. The initial divergence gave rise to Clade D, with a crown age estimated at 4.27 Ma, followed by Clade C, with a crown age estimated at 3.39 Ma. The four clades were not showed a clear spatial distribution. Suitable climatic conditions for the species were identified, including warmest quarter precipitation 433.20 mm ~ 1,524.07 mm, driest month precipitation > 12.06 mm, and coldest month min temp > -4.34 °C. The high suitability distribution showed contraction in LIG to LGM, followed by expansion from LGM to present. The Hengduan Mountains acted as a glacial refuge for the species during climate changes. Conclusions Our findings reflected a clear phylogenetic relationships and divergence within species L. japonicus and the identified hotspot regions could facilitate the genotype discrimination. The divergence time estimation and suitable area simulation revealed evolution dynamics of this species and may propose conservation suggestions and exploitation approaches in the future.

(Chinese motherwort) is a medicinal Lamiaceae species renowned for its pharmacological compounds, yet its mitochondrial genome remains unexplored. Elucidating mitogenomic structure and evolution can inform plant genetics, phylogenetics, and molecular breeding. We assembled the complete mitochondrial genome of using a combination of Oxford Nanopore long reads and Illumina short reads. Three assembly strategies- assembly with PMAT and Flye, and hybrid assembly with Unicycler-were integrated and validated via read mapping and comparison to reference mitogenomes ( ). Annotation employed GeSeq, tRNAscan-SE, and manual curation. Repeat elements (SSR, tandem, dispersed) were identified with MISA, TRF, and REPuter; plastid-to-mitochondrion transfers (MTPTs) were detected by BLASTN against the assembled plastome; and RNA editing sites were predicted using Deepred-mt. Phylogenetic and synteny analyses were conducted with IQ-TREE, MAFFT alignments of 24 conserved PCGs, and NGenomeSyn visualization. The circular mitogenome spanned 384,199 bp (45.1% GC) and encoded 35 protein-coding genes, 11 tRNAs, and 3 rRNAs. We detected 241 SSRs, 13 tandem repeats, and 90 dispersed repeats, indicating extensive recombination potential. Thirty-one MTPTs totaling 24,818 bp (6.46% of the mitogenome) were identified. Comparative analyses revealed strong purifying selection (Ka/Ks < 1) across most PCGs, with selective signatures in atp4 and ccmB. Phylogenetic inference placed among Lamiales, closely allied to and . Synteny maps demonstrated frequent genome rearrangements. Deepred-mt predicted 408 C-to-U RNA editing sites, notably in nad4 and , including novel start and stop codons. The mitogenome exhibits marked structural plasticity, reflecting dynamic repeats and organelle-to-organelle DNA transfers. Extensive RNA editing underscores post-transcriptional regulation in mitochondrial function. These findings enrich genomic resources for , support phylogenetic and evolutionary studies in Lamiaceae, and lay groundwork for molecular breeding and conservation strategies targeting mitochondrial traits.

Leonurus sibiricus L. (LS) is a medicinal plant used in East Asia, Europe and the USA. LS is primarily used in the treatment of gynecological diseases, and recent studies have demonstrated that it exerts anti-inflammatory and antioxidant effects. To the best of our knowledge, the present study demonstrated for the first time that LS may promote osteoblast differentiation and suppress osteoclast differentiation in vitro, and that it inhibited lipopolysaccharide (LPS)-induced bone loss in a mouse model. LS was observed to promote the osteoblast differentiation of MC3T3-E1 cells and upregulate the expression of runt-related transcription factor 2 (RUNX2), a key gene involved in osteoblast differentiation. This resulted in the induction of the expression of various osteogenic genes, including alkaline phosphatase (ALP), osteonectin (OSN), osteopontin (OPN), type I collagen (COL1) and bone sialoprotein (BSP). LS was also observed to inhibit osteoclast differentiation and bone resorption. The expression levels of nuclear factor of activated T-cells 1 (NFATc1) and c-Fos were inhibited following LS treatment. NFATc1 and c-Fos are key markers of osteoclast differentiation that inhibit receptor activator of nuclear factor-κB ligand (RANKL)-induced mitogen-activated protein kinase (MAPKs) and nuclear factor (NF)-κB. As a result, LS suppressed the expression of osteoclast-associated genes, such as matrix metallopeptidase-9 (MMP-9), cathepsin K (Ctsk), tartrate-resistant acid phosphatase (TRAP), osteoclast-associated immunoglobulin-like receptor (OSCAR), c-src, c-myc, osteoclast stimulatory transmembrane protein (OC-STAMP) and ATPase H+ transporting V0 subunit d2 (ATP6v0d2). Consistent with the in vitro results, LS inhibited the reduction in bone mineral density and the bone volume/total volume ratio in a mouse model of LPS-induced osteoporosis. These results suggest that LS may be a valuable agent for the treatment of osteoporosis and additional bone metabolic diseases.

Linnaeus 1753, an annual or biennial herb found in northern China, Mongolia, and Russia, typically grows in stony, sandy grasslands, and pine forests. This study sequenced and reported the complete chloroplast genome of for the first time. The entire circular genome measures 151,689 bp in length, with a GC content of 38.4%. A total of 133 genes were annotated, including 88 protein-coding genes, 37 tRNAs, and eight rRNAs. The genome exhibits a typical quadripartite structure, comprising a large single-copy (LSC 82,820 bp) region, a small single-copy (SSC 17,619 bp) region, and a pair of inverted repeat (IR 25,625 bp each) regions. Phylogenetic analysis using the maximum-likelihood method indicates that is most closely related to Houttuyn. This study provides valuable genomic resources for further research on the phylogenetics and biodiversity of the genus Leonurus.