BMC Genomics最新文献

{"title":"A tritrophic plant-insect-pathogen system used to develop a closely linked Rag2 and Rsv1-h recombinant haplotype in double-resistant soybean germplasm.","authors":"Luis G Posadas, Edson Ll Baldin, Lia Marchi-Werle, Tiffany M Heng-Moss, Scott Speck, Robert M Stupar, Kent M Eskridge, George L Graef","doi":"10.1186/s12864-025-11686-8","DOIUrl":"https://doi.org/10.1186/s12864-025-11686-8","url":null,"abstract":"<p><strong>Background: </strong>The colocalization of two resistance (R) genes on chromosome 13 of soybean (Glycine max (L.) Merrill) that confer resistance against the soybean aphid (Aphis glycines) and soybean mosaic virus (SMV) gives rise to a very unique R-avr tritrophic incompatible interaction system that goes across biological kingdoms. In this tritrophic system, the insect is the only natural vector of the virus and soybean is a host-plant for both pests/pathogen. The almost unavoidable co-evolution of pathogen-vector with that of the R-genes in soybean plants through an endless arms race to avoid each other's defense-attack mechanisms raises interesting questions. The objectives of this work were to (i) develop double-resistant recombinant inbred lines (RILs) with a Rag2-Rsv1-h gene haplotype in coupling phase using resistance alleles from two different genetic sources (PI 243540 (Rag2) and Suweon 97 (Rsv1-h)), (ii) confirm phenotypically the resistant reaction against both pests in double-resistant RILs, and (iii) dissect the Rag2-Rsv1-h region with molecular markers and investigate the potential for structural variation.</p><p><strong>Results: </strong>We observed a recombination event in identified double-resistant F<sub>3:5</sub> RILs in a region of chromosome 13 ca. 21 kb long (between positions 30,297,227 and 30,318,949 in Wm82.a2.v1) that lies between the reported locations of the Rsv1-h and Rag2 genes (29,815,463--29,912,369 and 30,412,581--30,466,533 intervals, respectively, based on Wm82.a2.v1), indicating the double-resistant haplotype is in coupling phase. The tight LD estimates obtained between haplotype markers underscored the physical proximity of the two resistance genes. Only 10 recombinant haplotype classes (excluding double heterozygotes) were observed among the 51 that were possible with a four loci haplotype. The 10 recombinant classes represented 15 out of 192 screened individuals. A joint SMV-aphid phenotypic greenhouse screen allowed us to identify the best aphid biotype 1 and SMV-G1, double resistant haplotype class in recombinant progeny. Our molecular marker results agree with previous fine-mapping reports and preclude the presence of resistance genes other than Rag2 and Rsv1-h in double-resistant RILs. A comparative genomic hybridization analysis revealed no obvious structural variants in the region.</p><p><strong>Conclusions: </strong>To our knowledge, this is the first report of double-resistant Rag2-Rsv1-h soybean RILs that used a plant-insect-pathogen tritrophic system for germplasm enhancement. The co-occurrence of Rag and Rsv genes in a region that clusters resistance genes on chromosome 13 may be a unique feature of domesticated soybean. The recombinant genotypes will be useful in breeding to develop soybean cultivars with resistance to both the vector and the virus. The parental and recombinant genotypes may be helpful in future studies to elucidate interesting evolutionary questions regarding vector, ","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"539"},"PeriodicalIF":3.5,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144156872","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A probabilistic approach to visualize the effect of missing data on PCA in ancient human genomics.","authors":"Susanne Zabel, Samira Breitling, Cosimo Posth, Kay Nieselt","doi":"10.1186/s12864-025-11728-1","DOIUrl":"https://doi.org/10.1186/s12864-025-11728-1","url":null,"abstract":"<p><strong>Background: </strong>Principal Component Analysis (PCA) is widely used in population genetics to visualize genetic relationships and population structures. In ancient genomics, genotype information may in parts remain unresolved due to the low abundance and degraded quality of ancient DNA. While methods like SmartPCA allow the projection of ancient samples despite missing data, they do not quantify projection uncertainty. The reliability of PCA projections for often very sparse ancient genotype samples is not well understood. Ignoring this uncertainty may lead to overconfident conclusions about the observed genetic relationships and population structure.</p><p><strong>Results: </strong>This study systematically investigates the impact of missing loci on PCA projections using both simulated and real ancient human genotype data. Through extensive simulations with high-coverage ancient samples, we demonstrate that increasing levels of missing data can lead to less accurate SmartPCA projections, highlighting the importance of considering uncertainty when interpreting PCA results from ancient samples. To address this, we developed a probabilistic framework to quantify the uncertainty in PCA projections due to missing data. By applying our methodology to modern and ancient West Eurasian genotype samples from the Allen Ancient DNA Resource database, we could show a high concordance between our predicted projection and empirically derived distributions. Applying this framework to real-world data, we demonstrate its utility in predicting and visualizing embedding uncertainties for ancient samples of varying SNP coverages.</p><p><strong>Conclusion: </strong>Our results emphasize the importance of accounting for projection uncertainty in ancient population studies. We therefore make our probabilistic model available through TrustPCA, a user-friendly web tool that provides researchers with uncertainty estimates alongside PCA projections, facilitating data exploration in ancient human genomic studies and enhancing transparency in data quality reporting.</p>","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"537"},"PeriodicalIF":3.5,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144156892","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of functional markers for peroxidase activity genes TaPod-7AS and TaPod-4AL in wheat.","authors":"Xinyuan Liu, Yukun Cheng, Hongwei Geng, Yi Ren, Junmei Cao, Bin Bai, Yating Xue","doi":"10.1186/s12864-025-11597-8","DOIUrl":"https://doi.org/10.1186/s12864-025-11597-8","url":null,"abstract":"<p><p>Grain peroxidase (POD) has a pivotal role in determining wheat flour color and end-use products. High POD activity in wheat grains can improve the whiteness and commercial value of flour. The POD genes TaPod-7AS and TaPod-4AL have high effects on POD activity in wheat. Function marker for POD genes can be used to improve flour color attributes accurately and efficiently in wheat molecular breeding. Here, wheat POD genes TaPod-7AS and TaPod-4AL were sequenced and characterized, gene expression analysis indicated that TaPod-7AS, TaPod-4AL, TaPod-D1 genes were highly expressed at 21 days after flowering. Function markers POD-7AS1 and POD-7AS2 were developed for TaPod-7AS, amplifying 424-bp and 464-bp PCR fragments in the cultivars with low and high POD activities, respectively. The markers POD-4AL1 and POD-4AL2 for TaPod-4AL amplified 541-bp and 391-bp fragments in the cultivars with low and high POD activities, respectively. The functional markers were validated in a diverse panel of 268 winter wheat cultivars and advanced lines, indicating significant associations between genotypes and POD activity (P < 0.05). The genotypes with favorable alleles TaPod-7ASb/TaPod-4ALb/TaPod-D1b had higher POD activity (749.9 U.g^-1.min^-1) than those with alleles TaPod-7ASa/TaPod-4ALa/TaPod-D1a (mean POD activity 619.4 U.g^-1.min^-1).</p>","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"538"},"PeriodicalIF":3.5,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144156873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptome analysis reveals age-specific growth characteristics in the rumen of Hanwoo (Korean native cattle) steers.","authors":"Yejee Park, Sunsik Jang, Han-Ha Chai, Ju-Whan Son, Dajeong Lim, Woncheoul Park","doi":"10.1186/s12864-025-11712-9","DOIUrl":"https://doi.org/10.1186/s12864-025-11712-9","url":null,"abstract":"<p><strong>Background: </strong>Hanwoo cattle are a Korean breed renowned for their cultural significance and high-quality beef, characterized by low cholesterol and a high unsaturated fat ratio. Their growth is divided into a growing stage focused on development and a fattening stage for marbling. Proper feed management, considering genetic and environmental factors, is vital for maximizing growth potential. The rumen plays a crucial role in digestion and gene expression regulation, with rumen fermentation being central to nutrient absorption and cattle health. In this study, we conduct a transcriptome analysis of the rumen at eight timepoints. Our goal is to identify genetic factors that influence the growth of Hanwoo steers to enhance our understanding of the rumen's functions during Hanwoo growth.</p><p><strong>Results: </strong>In the RNA-sequencing analysis of Hanwoo steer rumen, differential gene expression was examined over eight timepoints, highlighting significant genetic changes, particularly between 12 and 26 months. The results of a weighted gene co-expression network analysis were identified and organized into three modules: turquoise, blue, and yellow. The turquoise module, linked to immune response, showed significantly down-regulation in genes at 30 months. The blue module, associated with steroid metabolism, was notably up-regulated at 26 months. The yellow module's genes showed a consistent increase in expression with growth. These modules and their functional annotations provide a deeper understanding of the biological processes during Hanwoo growth, highlighting the intricate relationship between gene expression and cattle development.</p><p><strong>Conclusions: </strong>The growth stages of Hanwoo steers were explored in our investigation utilizing rumen transcriptome data. The rumen plays a critical role in their development, particularly during the growing and fattening stages. Proper feed management, considering the rumen's function, is essential for optimal growth. Transcriptome analysis helps identify genes associated with growth and provides insights for cattle breeding and management practices. Understanding the complex connection between gene expression and Hanwoo development is essential for maximizing productivity and health.</p>","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"530"},"PeriodicalIF":3.5,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-quality assembly of the chromosomal genome for Flemingia macrophylla reveals genomic structural characteristics.","authors":"Ye Ding, Yi Zhao, Yangqin Xie, Fan Wang, Wu Bi, Mengyao Wu, Guilin Zhao, Yun Gong, Wenli Li, Peng Zhang","doi":"10.1186/s12864-025-11705-8","DOIUrl":"https://doi.org/10.1186/s12864-025-11705-8","url":null,"abstract":"<p><p>Flemingia macrophylla, a prominent shrub species within the Fabaceae family, is widely distributed across China and Southeast Asia. In addition to its ecological importance, it possesses notable medicinal value, with its roots traditionally used for treating rheumatism, enhancing blood circulation, and alleviating joint pain. We employed Nanopore sequencing platforms to generate a high-quality reference genome for F. macrophylla, with an assembled genome size of 1.01 Gb and a contig N50 of 59.43 Mb. A total of 33,077 protein-coding genes were predicted, and BUSCO analysis indicated a genome completeness of 99%. Phylogenomic analyses showed that F. macrophylla is most closely related to Cajanus cajan among the sampled taxa, with an estimated divergence time of 13.2-20.0 MYA. Evidence of whole-genome duplication (WGD) events was detected in F. macrophylla, C. cajan, and P. vulgaris, with these species sharing two WGD events. The unique gene families in F. macrophylla are associated with strong resistance to both abiotic and biotic stress, supporting its remarkable ecological adaptability. Furthermore, gene family expansion analysis revealed a significant enrichment of genes related to secondary metabolites biosynthesis, providing a molecular basis for its high medicinal value. In summary, this study provides a foundational genomic resource for F. macrophylla, offering valuable insights into its genetic architecture, evolutionary history, and potential applications in medecine and agriculture. The comprehensive analyses lay the groundwork for future research into the species's medicinal properties and evolutionary biology.</p>","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"535"},"PeriodicalIF":3.5,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149159","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stage-specific transcriptomic analysis reveals insights into the development, reproduction and biological function of allergens in the European house dust mite Dermatophagoides pteronyssinus.","authors":"José Cristian Vidal-Quist, Félix Ortego, Bart N Lambrecht, Stephane Rombauts, Pedro Hernández-Crespo","doi":"10.1186/s12864-025-11703-w","DOIUrl":"https://doi.org/10.1186/s12864-025-11703-w","url":null,"abstract":"<p><strong>Background: </strong>House dust mites (HDMs) such as Dermatophagoides pteronyssinus are major allergy elicitors worldwide, yet their gene expression across developmental stages remains underexplored. Herein, we report a comprehensive RNAseq analysis of larvae, nymphs, and adult males and females, mapped to a recently published high-quality genome with extended functional annotations.</p><p><strong>Results: </strong>Analysis of differentially expressed genes (DEG) revealed that female-biased expression was the most prevalent profile (16% of genes), while males exhibited the highest fold-change differences. DEG data, combined with network clustering and functional enrichment analysis, highlighted distinct genes and biological processes for each stage and sex: females showed upregulation of genes related to cell division and oogenesis, with vitellogenins among the most abundant transcripts; males exhibited increased expression of genes encoding putative seminal fluid proteins (e.g. endopeptidases, serpins, antimicrobial peptides), and those involved in reproductive regulation (e.g. testis-specific serine kinases); while juveniles displayed enhanced expression of genes related to energy metabolism and growth. Further analysis of endocrine pathways revealed non-canonic mechanisms compared to insect models, particularly in ecdysteroid and sesquiterpenoid biosynthesis and regulation. Expression patterns in genes involved in cuticle formation were also identified, reflecting their role in developmental transitions and sexual differentiation. Allergen and allergen-related gene expression showed an overall increase in feeding juveniles, as well as sex-biased expression, with Der p 27 upregulated in females. These findings provide insight into the physiological roles of allergens in digestion, immunity, and muscle formation, among other functions. Additionally, seven new horizontally transferred genes, including a DNA-repair photolyase linked to females, and novel multigene families (e.g. 119 male-specific beta-propeller proteins, 70 hypothetical cuticular proteins, 23 tetraspanin-like proteins, 5 female-associated putative odorant-binding proteins) were identified.</p><p><strong>Conclusions: </strong>This study provides the first genome-wide transcriptomic analysis of a HDM across life stages and sexes, expanding our understanding of the molecular mechanisms underlying mite development, sexual reproduction, and allergen expression. The generated data, fully available via supplementary spreadsheet and the ORCAE online platform, provide a valuable foundation for future allergy research and the development of new mite control strategies.</p>","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"527"},"PeriodicalIF":3.5,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149245","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction: Genome-wide identification and expression analysis of the JMJ-C gene family in melon (Cucumis melo L.) reveals their potential role in fruit development.","authors":"Wuyun Jin, Wei Yan, Ming Ma, Agula Hasi, Gen Che","doi":"10.1186/s12864-025-11704-9","DOIUrl":"https://doi.org/10.1186/s12864-025-11704-9","url":null,"abstract":"","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"533"},"PeriodicalIF":3.5,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149229","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative genomic analysis of immune-related genes and chemosensory receptors provides insights into the evolution and adaptation of four major domesticated Asian carps.","authors":"Ling Shi, Judy Kin-Wing Ng, Qing Xiong, Kelvin Fu-Kiu Ao, Soo-Kyung Shin, Cherie Tsz-Yiu Law, Weixue Mu, Guang-Ming Liu, Shitao Rao, Stephen Kwok-Wing Tsui","doi":"10.1186/s12864-025-11719-2","DOIUrl":"https://doi.org/10.1186/s12864-025-11719-2","url":null,"abstract":"<p><strong>Background: </strong>Ctenopharyngodon idella (grass carp), Mylopharyngodon piceus (black carp), Hypophthalmichthys nobilis (bighead carp), and Hypophthalmichthys molitrix (silver carp), collectively known as the four major domesticated Asian carp, are freshwater fish species from the family Cyprinidae and are widely consumed in China. Current studies on these species primarily focus on immune system regulation and the growth and development of individual species. However, in-depth genomic investigations and comprehensive comparative analysis remained limited.</p><p><strong>Methods: </strong>The complete genomes of Ctenopharyngodon idella, Mylopharyngodon piceus and Hypophthalmichthys nobilis were assembled using a hybrid approach that integrated both next- and third-generation sequencing reads, followed by annotation using the MAKER2 pipeline. Based on the high-quality genomes of Ctenopharyngodon idella, Mylopharyngodon piceus Hypophthalmichthys nobilis, and Hypophthalmichthys molitrix, a comparative genomic analysis was conducted using bioinformatic tools to investigate gene family evolution in these four domesticated Asian carp species.</p><p><strong>Results: </strong>High-quality genomes of Ctenopharyngodon idella, Mylopharyngodon piceus, and Hypophthalmichthys nobilis were assembled, achieving over 90% completeness. Immune-related gene families, including MHC class I and NLRC3-like genes, have undergone rapid evolution, with Ctenopharyngodon idella exhibiting significant expansion of NLRC3-like genes. Massive tandem duplication events were identified in trace amine-associated receptors (TAARs), and rapid expansion was observed in TAAR16 and TAAR29. Additionally, a novel TAAR gene cluster was identified in all four Asian carp species. Comparative genomic analysis revealed the expansion of type 1 taste receptor genes, particularly in Ctenopharyngodon idella and Mylopharyngodon piceus.</p><p><strong>Conclusion: </strong>This study has successfully constructed the high-quality genomes of Ctenopharyngodon idella, Mylopharyngodon piceus, and Hypophthalmichthys nobilis. The comparative genomic analysis revealed the evolution of immune-related genes and chemosensory receptors in the four major domesticated Asian carp species. These findings suggested the enhanced immunity and sensory perception in these species, providing valuable insights into their adaptation, survival and reproduction.</p>","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"529"},"PeriodicalIF":3.5,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149225","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transcriptomic analysis of wild Cannabis sativa: insights into tissue- and stage-specific expression and secondary metabolic regulation.","authors":"Jinyuan Hu, Zishi Wang, He Xu, Zhenlong Wang, Ning Li, Rui Feng, Jianyu Yin, Fangru Liu, Baishi Wang","doi":"10.1186/s12864-025-11697-5","DOIUrl":"https://doi.org/10.1186/s12864-025-11697-5","url":null,"abstract":"<p><p>Cannabis sativa is a medicinally and economically significant plant known for its production of cannabinoids, terpenoids, and other secondary metabolites. This study presents a transcriptomic analysis to elucidate tissue-specific expression and regulatory mechanisms across leaves, stems, and roots. A total of 2,530 differentially expressed genes (DEGs) were identified, with key genes such as terpene synthase (TPS) and phenylalanine ammonia-lyase (PAL) exhibiting elevated expression in leaf tissues, emphasizing their roles in terpenoid and phenylpropanoid biosynthesis. Alternative splicing (AS) analysis revealed 8,729 distinct events, dominated by exon skipping, contributing to transcriptomic diversity. Long non-coding RNA (lncRNA) prediction identified 3,245 candidates, many of which displayed tissue-specific expression patterns and co-expression with metabolic genes, suggesting regulatory roles in secondary metabolism. Additionally, 12,314 SNPs and 2,786 INDELs were detected, with notable enrichment in genes associated with secondary metabolite biosynthesis, particularly in leaf tissues. These findings advance the understanding of molecular mechanisms governing secondary metabolism and genetic diversity in C. sativa, providing valuable insights for future metabolic engineering and breeding strategies to enhance cannabinoid production.</p>","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"528"},"PeriodicalIF":3.5,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149190","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Mg²⁺-dependent high-yield method for extracting high-molecular-weight genomic DNA from a single planarian specimen.","authors":"Ao Li, Bingrui Sun, Ying Zhang, Ping Yu, Jicheng Qu, Hongkuan Deng, Qiuxiang Pang, Fengtang Yang","doi":"10.1186/s12864-025-11731-6","DOIUrl":"https://doi.org/10.1186/s12864-025-11731-6","url":null,"abstract":"<p><strong>Background: </strong>The isolation of intact, high-molecular-weight genomic DNA (HMW gDNA) is essential for achieving complete genome assemblies. However, extracting HMW gDNA from a single individual of Dugesia japonica remains a technical challenge using the standard protocol, probably due to the presence of abundant polysaccharides and nucleases.</p><p><strong>Results: </strong>In this study, we have developed a more robust protocol for preparing HMM gDNA, with high yields and quality, from a single D. japonica. The key step in our protocol involves the use of a Mg²⁺-dependent lysis buffer, rather than using metal cation chelation to block the activities of DNase I as in the standard protocol. Using this approach were able to achieve a yield of about 10-15 µg of HWM gDNA per worm. Our method showed species- and region-specific effectiveness, with optimal results observed at 20 mM Mg²⁺ for our local D. japonica specimens. The extracted HMW gDNA is fully compatible with advanced long-read sequencing platforms such as PacBio HiFi and Oxford Nanopore. However, when applied to Schmidtea mediterranea and D. japonica specimens from Beijing, the method was ineffective and led to progressive gDNA degradation.</p><p><strong>Conclusions: </strong>This protocol offers a simple and high-yield solution for isolating HMW gDNA from D. japonica. It also provides an alternative for organisms whose gDNA consistently exhibits unexplained degradation using established protocols.</p>","PeriodicalId":9030,"journal":{"name":"BMC Genomics","volume":"26 1","pages":"532"},"PeriodicalIF":3.5,"publicationDate":"2025-05-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144149213","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}