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13 pages, 973 KB  
Article
Fine Mapping of McFFFN4.1 and Identification of a Candidate Gene and KASP Marker for the First Female Flower Node in Bitter Gourd
by Xiaoxi Liu, Ming He, Yangyi Zheng, Jianning Luo, Junxing Li, Hao Gong, Haibin Wu, Gangjun Zhao, Liting Deng, Xueting Wang, Chengcheng Feng and Xiaoming Zheng
Horticulturae 2026, 12(5), 520; https://doi.org/10.3390/horticulturae12050520 (registering DOI) - 23 Apr 2026
Abstract
The first female flower node (FFFN) is a crucial trait affecting earliness and yield in bitter gourd (Momordica charantia L.). To identify the genetic locus and candidate gene controlling FFFN, we performed phenotypic and genetic analyses using two parental lines, ‘M144’ (average [...] Read more.
The first female flower node (FFFN) is a crucial trait affecting earliness and yield in bitter gourd (Momordica charantia L.). To identify the genetic locus and candidate gene controlling FFFN, we performed phenotypic and genetic analyses using two parental lines, ‘M144’ (average FFFN: 6.3 ± 2.0) and ‘K55’ (average FFFN: 22.0 ± 4.5), along with their F1 hybrid and an F2 population consisting of 317 individuals. The results show that the low FFFN trait was incompletely dominant over the high FFFN trait. Using BSA-seq, we mapped a FFFN locus to an interval of 18.8–22.5 Mb on chromosome 4. Fine mapping with KASP markers narrowed the McFFFN4.1 to a 73.05 kb interval between markers 25QP334 and 26QP20, which contained seven predicted genes. Transcriptome analysis revealed that only Moc04g29650, which is annotated as cytochrome b-c1 complex subunit Rieske, was differentially expressed between the parents within this mapping interval. Sequence comparison identified a single SNP (C > A) in the promoter region of Moc04g29650, which was located within a putative YAB1/FIL-binding motif. Given the known role of FILAMENTOUS FLOWER (FIL) in regulating floral transition in Arabidopsis thaliana, Moc04g29650 is proposed as the most likely candidate gene for McFFFN4.1. The KASP marker 26QP20, located near Moc04g29650, showed the strongest association with FFFN in the F2 population, with a maximum LOD score of 5.45, and thus represents a valuable tool for marker-assisted selection (MAS) breeding in bitter gourd. This study lays a foundation for cloning McFFFN4.1 and genetically improving early maturity in bitter gourd. Full article
10 pages, 3576 KB  
Article
Effects of Single-Nucleotide Polymorphisms on the Estimated Breeding Values for Feet in Holstein-Friesian Cows in Hungary
by Attila Zsolnai, László Bognár, Szabolcs Albin Bene, Zsolt Jenő Kőrösi, László Rózsa, Ferenc Szabó and István Anton
Animals 2026, 16(9), 1299; https://doi.org/10.3390/ani16091299 - 23 Apr 2026
Abstract
The aim of this study was to identify SNPs in the cattle genome associated with estimated breeding values of feet (EBVfeet) in Holstein-Friesian (HF) cows in Hungary. Foot health is of major importance in dairy cattle industry whereas claw disorders are [...] Read more.
The aim of this study was to identify SNPs in the cattle genome associated with estimated breeding values of feet (EBVfeet) in Holstein-Friesian (HF) cows in Hungary. Foot health is of major importance in dairy cattle industry whereas claw disorders are leading to lameness and thus result in low fertility rates and productivity. Genotyping was performed using the EuroG_MDv4 microarray platform. The final database comprised 2963 animals and 59,151 SNPs. EBVfeet values have been divided into high and low groups. All calculations regarding the genetic differentiation (genome-wide and locus-specific) between high- and low-value groups for EBVfeet, linear regression, and haplotype association tests have been performed with the SNP and Variation Suite software. Thirty-nine SNPs associated with EBVfeet were determined on BTAs 3, 7, 8, 15, 21, and X. The maximum values of the identified SNPs were 0.22 for Fst_marker, 23.1 for the −log10(p) of the linear regression, and 26.3 for the −log10(p) of the haplotype association tests on BTA 3. The closest genes to SNPs associated with estimated breeding values for feet (EBVfeet) are mainly associated with tissue structure, immune response, metabolism, growth, development, transport and signaling. Our results could add additional information to the genetic programs focusing on the improvement of foot health in HF cattle. Full article
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18 pages, 10656 KB  
Article
An NS1-F161L Substitution Determines Host-Driven Virulence Enhancement of H5N6 Avian Influenza Virus in Ducks
by Yuwei Wu, Zhifan Li, Nuo Xu, Zijun Lu, Yurui Dong, Kunlin Li, Ying Bian, Chenzhi Huo, Tao Qin, Sujuan Chen, Hui Yang, Daxin Peng and Xiufan Liu
Viruses 2026, 18(5), 488; https://doi.org/10.3390/v18050488 - 23 Apr 2026
Abstract
H5 subtype avian influenza virus (AIV) can infect both chickens and ducks, leading to substantial economic losses. Nevertheless, certain strains cause silent infections in ducks. In this study, a goose-origin clade 2.3.4.4h H5N6 AIV was isolated, which caused high mortality in mixed-gender white [...] Read more.
H5 subtype avian influenza virus (AIV) can infect both chickens and ducks, leading to substantial economic losses. Nevertheless, certain strains cause silent infections in ducks. In this study, a goose-origin clade 2.3.4.4h H5N6 AIV was isolated, which caused high mortality in mixed-gender white leghorn chickens but no deaths in mixed-gender mallard ducks. After independent serial in vitro passage in duck embryo fibroblasts (DEFs) and in vivo passage in specific-pathogen-free (SPF) ducks, the DEF-passage 10 (P10) virus induced markedly higher mortality rates and viral loads in SPF ducks compared to the DEF-P1 virus and the original parental virus prior to passage. Similarly, the in vivo-passaged P3 and P4 viruses exhibited significantly higher mortality rates than the P1 virus in SPF ducks, with 100% mortality and markedly increased viral titers in the organs. A whole-genome SNP analysis identified seven high-frequency mutations in the M1, NA and NS1 proteins. The NS1-F161L substitution virus exhibited significantly increased mortality rates, viral loads in multiple tissues, and a robustly induced innate immune response in ducks. Furthermore, dynamic evolutionary variations in the NS1 protein among global H5 avian influenza viruses revealed that the NS1-F161L substitution became dominant in clade 2.3.4.4b viruses in 2021 and subsequent years. Collectively, our findings demonstrate that host-driven adaptation can rapidly increase the pathogenicity of H5N6 AIVs in ducks and identify NS1-F161L as a critical virulence marker. These results offer novel insights relevant to the molecular surveillance, virulence prediction, and risk assessment of circulating H5 AIVs in waterfowl. Full article
(This article belongs to the Special Issue Avian Viruses and Antiviral Immunity)
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13 pages, 1161 KB  
Article
A Quantitative Trait Nucleotide-Based Genomic Selection Strategy for Seed Oil and Protein Content in Soybean
by Guang Li, Huangkai Zhou, Javaid Akhter Bhat, Kuanqiang Tang, Jiantian Leng, Xianzhong Feng, Xiangfeng Wang and Suxin Yang
Plants 2026, 15(9), 1296; https://doi.org/10.3390/plants15091296 - 22 Apr 2026
Abstract
In recent years, genomic selection (GS) has been widely adopted in plant breeding; however, its practical application is constrained by the high cost of genotyping large segregating populations. To address this issue, this study employed a Quantitative Trait Nucleotide (QTN)-assisted GS strategy to [...] Read more.
In recent years, genomic selection (GS) has been widely adopted in plant breeding; however, its practical application is constrained by the high cost of genotyping large segregating populations. To address this issue, this study employed a Quantitative Trait Nucleotide (QTN)-assisted GS strategy to evaluate its efficiency in reducing genotyping costs for soybean seed oil content (OC) and protein content (PC). Based on six multi-parent F4 populations (n = 4404) derived from seven elite soybean cultivars, which were genotyped using a 20K SNP chip, we identified 83 and 110 QTNs that were significantly associated with OC and PC, respectively. Among these loci, 37 and 62 QTNs were specific to OC and PC, respectively. Genomic prediction accuracies were evaluated across different training population (TP) sizes using three marker panels: genome-wide SNPs, all detected QTNs, and trait-specific QTNs. The panel consisting of all detected QTNs exhibited significantly higher prediction accuracy than the other two panels, except for PC when using 90% of the population as the training set. Phenotypic verification of the selected individuals showed that the PC-specific QTN panel yielded higher PC values and increased OC + PC values compared with the other marker panels. These results demonstrate that a small set of QTNs provides a cost-effective approach for genomic selection in practical soybean breeding programs. Full article
(This article belongs to the Special Issue Genetic Improvement of Oilseed Crops)
13 pages, 1237 KB  
Article
Development of a Medium-Density Genotyping Platform to Accelerate Genetic Gain in Fresh Edible Maize
by Jingtao Qu, Diansi Yu, Wei Gu, Yingjie Zhao, Kai Li, Hui Wang, Pingdong Sun, Felix San Vicente, Xuecai Zhang, Ao Zhang, Hongjian Zheng and Yuan Guan
Plants 2026, 15(9), 1288; https://doi.org/10.3390/plants15091288 - 22 Apr 2026
Abstract
Genotyping is a key step in molecular breeding. Due to its cost-effectiveness, accuracy, and flexibility, genotyping by target sequencing (GBTS) has become a preferred technology for medium-density genotyping. In this study, a new GBTS array for fresh edible maize was developed using resequencing [...] Read more.
Genotyping is a key step in molecular breeding. Due to its cost-effectiveness, accuracy, and flexibility, genotyping by target sequencing (GBTS) has become a preferred technology for medium-density genotyping. In this study, a new GBTS array for fresh edible maize was developed using resequencing data from 477 lines. The array contains 5759 SNPs evenly distributed across the maize genome, with average minor allele frequency (MAF) and polymorphism information content (PIC) values of 0.40 and 0.36, respectively. These SNPs are closely associated with 1566 functional genes. Cluster analysis of 198 maize lines based on the GBTS array was consistent with their pedigree relationships. Furthermore, 277 fresh waxy maize lines were genotyped and used for genomic selection analyses of hundred-kernel weight, kernel length, and kernel width. Comparative evaluation of different models indicated that Ridge Regression Best Linear Unbiased Prediction (rrBLUP) was the optimal model, with prediction accuracies of 0.33, 0.64, and 0.36, respectively. Additional analyses using different marker densities based on the rrBLUP model showed that prediction accuracy did not increase when the number of markers exceeded 2000, indicating that this array provides sufficient marker density for genetic analysis and genomic selection. Overall, this array provides a useful tool for genetic studies of fresh edible maize and facilitates the application of genomic selection in breeding programs. Full article
(This article belongs to the Section Plant Genetics, Genomics and Biotechnology)
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44 pages, 491 KB  
Article
Reaction of Wheat to European Virulence Races of Common Bunt (Tilletia spp.) and Mapping Race-Specific Resistance Genes Using SNP Markers
by Anders Borgen and Dennis Kjær Christensen
Plants 2026, 15(8), 1264; https://doi.org/10.3390/plants15081264 - 20 Apr 2026
Viewed by 431
Abstract
Common bunt of wheat (Tilletia spp.) remains a significant threat to wheat production in low-input and organic farming systems, where chemical seed treatments are restricted or avoided. Host resistance represents a key component of sustainable disease control, but its effective deployment requires [...] Read more.
Common bunt of wheat (Tilletia spp.) remains a significant threat to wheat production in low-input and organic farming systems, where chemical seed treatments are restricted or avoided. Host resistance represents a key component of sustainable disease control, but its effective deployment requires detailed knowledge of race-specific virulence and the genetic basis of resistance. In this study, we analysed the reaction of a large and diverse wheat germplasm collection to current European populations of common bunt and mapped the underlying resistance genes using SNP-based approaches. A total of 2731 wheat accessions were phenotyped from 2012 to 2025 using up to 42 purified bunt races with well-defined virulence profiles. Based on phenotypic responses to race-specific resistance patterns, accessions were grouped and compared with established differential lines. A total of 1504 selected accessions were genotyped using Illumina 26k SNP arrays, and resistance loci were identified by genome-wide association studies followed by fine mapping using recombination analysis. All classical Bt resistance genes from Bt1 to Bt10 and Bt13 and BtZ were mapped to defined physical intervals, and the genomic positions of 18 additional race-specific resistance genes were identified in a panel of germplasm. Our results confirm that several historically defined Bt genes, including Bt11 and Bt12, represent multi-gene resistance complexes rather than single loci. Also, genes established as separate genes may possibly be identical, including Bt4 being identical to Bt6, Bt10 being identical to BtZ, and Bt9 possibly being identical to one of the genes in the Bt11 complex. These findings highlight the need for a revised nomenclature of genes and a differential set of varieties. The identified resistance haplotypes provide an improved tool for marker-assisted selection and support the development of wheat cultivars with durable resistance to common bunt. Full article
(This article belongs to the Special Issue Wheat Breeding for Disease Resistance)
20 pages, 1643 KB  
Article
Genome-Wide Association Study and Candidate Gene Identification for Resistance to Bacterial Stem and Root Rot in Sweetpotato
by Xiangsheng Lin, Xiawei Ding, Shixu Zhou, Hongda Zou, Zhangying Wang, Xuelian Liang, Xiangbo Zhang and Lifei Huang
Biology 2026, 15(8), 643; https://doi.org/10.3390/biology15080643 (registering DOI) - 19 Apr 2026
Viewed by 135
Abstract
Bacterial stem and root rot (BSRR), caused by Dickeya dadantii, poses a severe threat to global sweetpotato production, yet the genetic architecture underlying resistance remains elusive. To dissect these mechanisms, we conducted a high-resolution genome-wide association study (GWAS) on 135 diverse accessions, [...] Read more.
Bacterial stem and root rot (BSRR), caused by Dickeya dadantii, poses a severe threat to global sweetpotato production, yet the genetic architecture underlying resistance remains elusive. To dissect these mechanisms, we conducted a high-resolution genome-wide association study (GWAS) on 135 diverse accessions, integrating two-year field phenotyping with best linear unbiased prediction (BLUP) and 6.8 million single-nucleotide polymorphism (SNP) markers. This approach mapped nine quantitative trait loci (QTLs) exhibiting significant allelic dosage-dependent effects, with the major locus, qBSRR.6.1 was the primary discriminator between resistant and susceptible genotypes. Crucially, transcriptomic profiling within these loci revealed distinct expression patterns: IbTCP5 and IbERF003 (located in qBSRR.5.1 and qBSRR.6.2) were highly expressed in the susceptible cultivar ‘Xinxiang’ but suppressed in the resistant ‘Guangshu87’. Furthermore, BSRR challenge identified IbPUB4, IbKCS5, and IbLig1 as priority candidate genes involved in defense, with expression patterns suggesting roles in ubiquitin-mediated protein turnover, cuticular wax biosynthesis, and DNA repair, respectively. In stark contrast, IbPUB25 was constitutively upregulated in ‘Xinxiang’, potentially acting as a negative regulator of immunity via degradation of target proteins. These findings elucidate the polygenic, dosage-sensitive nature of BSRR resistance and prioritize specific targets for future functional characterization. Pyramiding favorable alleles of positive candidates while silencing potential negative regulators like IbPUB25 offers a promising avenue for developing durable, high-resistance sweetpotato varieties. Full article
(This article belongs to the Section Genetics and Genomics)
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13 pages, 1480 KB  
Article
Genome-Wide Association Study Suggests rrp44 is a Key Regulator of Growth Traits in Channel Catfish (Ictalurus punctatus)
by Shiyong Zhang, Hongyan Liu, Yongqiang Duan, Minghua Wang and Xiaohui Chen
Curr. Issues Mol. Biol. 2026, 48(4), 420; https://doi.org/10.3390/cimb48040420 - 18 Apr 2026
Viewed by 141
Abstract
Understanding the genetic architecture underlying growth variation is central to improving aquaculture species through genomic selection. Here, we performed a genome-wide association study (GWAS) on 303 individuals from a G2 breeding population of channel catfish (Ictalurus punctatus) using whole-genome resequencing [...] Read more.
Understanding the genetic architecture underlying growth variation is central to improving aquaculture species through genomic selection. Here, we performed a genome-wide association study (GWAS) on 303 individuals from a G2 breeding population of channel catfish (Ictalurus punctatus) using whole-genome resequencing data. After stringent quality control, 5.64 million high-confidence single nucleotide polymorphisms (SNPs) were retained for association analyses of two key growth traits—monthly weight gain (MWG) and body depth (BH). We identified 15 and 28 loci significantly associated with MWG and BH, respectively, with the majority concentrated on chromosome 20. Two SNPs (Chr20:14,657,971 and Chr20:14,658,012) located in exon 9 of the rrp44 gene were significantly associated with both traits. Functional annotation and enrichment analyses revealed that the rrp44 gene, encoding an exoribonuclease subunit of the RNA exosome complex, participates in mitotic spindle regulation and post-transcriptional RNA decay, processes critical for cellular growth and metabolic homeostasis. We propose that rrp44 may influence growth through the modulation of feeding rhythm and circadian regulation, providing a potential molecular basis for growth heterogeneity in channel catfish. These findings enrich our understanding of growth-related genomic variation and offer valuable molecular markers for precision breeding and genetic improvement of catfish. Full article
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
19 pages, 6929 KB  
Article
Genomic Signatures of Somatic Mutation and Selection Shape Distinct Clonal Lineages in Bougainvillea × buttiana ‘Miss Manila’ Bud Sport
by Hongyan Meng, Qun Zhou, Duchao Chen, Bayan Huang, Mingqiong Zheng and Wanqi Zhang
Genes 2026, 17(4), 471; https://doi.org/10.3390/genes17040471 - 17 Apr 2026
Viewed by 216
Abstract
Background/Objectives: Bud sports (somatic mutations) offer a quick way to develop new bougainvillea varieties by altering specific traits while keeping the desirable genetic background of the original cultivar. However, we still lack a comprehensive understanding of their genomic architecture and the molecular [...] Read more.
Background/Objectives: Bud sports (somatic mutations) offer a quick way to develop new bougainvillea varieties by altering specific traits while keeping the desirable genetic background of the original cultivar. However, we still lack a comprehensive understanding of their genomic architecture and the molecular mechanisms behind their formation. This study aimed to characterize the population genomic characteristics of bud sports derived from the commercial variety Bougainvillea × buttiana ‘Miss Manila’. Methods: We employed genotyping by sequencing (GBS) on 39 accessions, including 27 bud sports and 12 conventional varieties. Population genomic analyses, such as principal component analysis (PCA), phylogenetic reconstruction, ADMIXTURE, and diversity statistics (π, He, Tajima’s D), were performed on 64,810 high-quality SNPs. Genome-wide scans for differentiation (FST) and selective sweeps (XP-CLR) were also conducted. Results: Bud sports showed significantly lower genetic diversity (π and He) than conventional varieties, which matches their clonal origin. PCA, phylogenetic, and ADMIXTURE analyses (optimal K = 4) revealed clear genetic differentiation and distinct population structures between the two groups. The bud sport population possessed fewer private alleles and a less negative Tajima’s D value. Genomic scans identified regions under selection in bud sports, with functional annotation pointed to genes involved in ubiquitin-mediated proteolysis and RNA transport. Notably, Bou_119143 (UDP-rhamnose rhamnosyltransferase 1) showed a high mutation frequency specifically in bud sports. Conclusions: We provide the first population-genomic evidence that bud sports of ‘Miss Manila’ are genetically distinct clonal lineages, shaped by somatic mutation and selection. These findings support bud sports as efficient sources for germplasm innovation. The identified genomic regions and candidate genes lay a foundation for future marker-assisted selection and molecular breeding in bougainvillea. Full article
(This article belongs to the Topic Genetic Breeding and Biotechnology of Garden Plants)
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20 pages, 1217 KB  
Article
Molecular Labelling Tool for Cereal Genetic Resources Management Derived from Barley and Tetraploid Wheat Genebank-Genomics Projects
by Workie Zegeye, Amanda Burridge, Ajay Siluveru, Simon Orford, Liz Sayers, Richard Goram, Richard Horler, Gary Barker and Noam Chayut
Plants 2026, 15(8), 1219; https://doi.org/10.3390/plants15081219 - 16 Apr 2026
Viewed by 334
Abstract
Globally, 5.94 million accessions are conserved across 867 genebanks, of which 41.5% (2.47 million) are cereal crop accessions. Only a small portion of global germplasm diversity has been marker-genotyped or genome-sequenced. Accurate identification of genebank accessions is essential to improve the efficiency and [...] Read more.
Globally, 5.94 million accessions are conserved across 867 genebanks, of which 41.5% (2.47 million) are cereal crop accessions. Only a small portion of global germplasm diversity has been marker-genotyped or genome-sequenced. Accurate identification of genebank accessions is essential to improve the efficiency and effectiveness of global genebanking. It is crucial for preserving the legacy knowledge associated with the germplasm and for maintaining its value to current plant science and breeding efforts. Existing practices generally fall into two categories: either expensive and complex, or inefficient, labour-intensive, and inaccurate. The first relies on high-resolution genomic sequences or saturated markers, while the second relies on morphological comparisons of regenerated plants with historical records. We propose a genotyping method based on a minimal set of Single Nucleotide Polymorphism (SNP) markers and exemplify its use on a genebank scale. We identified a small, effective set of SNPs that can differentiate between the global diversity of genebank accessions of barley (Hordeum vulgare and Hordeum spontaneum) and tetraploid wheat collections (Triticum turgidum) maintained at the Germplasm Resources National Capability at the John Innes Centre, UK. This approach offers a straightforward, automatable, and inexpensive alternative to traditional genebank crop descriptors used during seed regeneration and distribution. By establishing the minimal genomic resolution needed to distinguish genetically distinct accessions, we show that as few as 24 and 25 carefully chosen SNP markers for barley and durum wheat, respectively, can effectively differentiate individual accessions. Unlike morphology-based identification, which can detect mislabelling or contamination but often cannot prevent or correct such errors, our SNP-based molecular labelling enables error correction and the retrieval of lost germplasm identity. This study highlights how accuracy and reliability in germplasm management can be improved without costly whole-genome sequencing or resource-intensive analysis. We discuss the impact of this method on enhancing quality assurance in genebanks and its broader usefulness for the user community. Full article
(This article belongs to the Section Plant Genetic Resources)
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19 pages, 6663 KB  
Article
Genetic Diversity and Core Germplasm Development in Yunnan Tartary Buckwheat Based on Phenotypic and SNP Data
by Bingxin Zhai, Daowang Sun, Chunyan Huang, Zihan Zhao, Jiaxing Xie, Xin Liu, Wenjie Lu, Guang Wang and Lihua Wang
Plants 2026, 15(8), 1197; https://doi.org/10.3390/plants15081197 - 14 Apr 2026
Viewed by 331
Abstract
Southwest China harbors the world’s richest germplasm resources of Tartary buckwheat (Fagopyrum tataricum). However, their effective utilization is severely constrained by poor management and narrow genetic diversity. Developing a core collection is a key strategy for overcoming these bottlenecks and facilitates [...] Read more.
Southwest China harbors the world’s richest germplasm resources of Tartary buckwheat (Fagopyrum tataricum). However, their effective utilization is severely constrained by poor management and narrow genetic diversity. Developing a core collection is a key strategy for overcoming these bottlenecks and facilitates the efficient conservation and utilization of germplasm resources. Therefore, we aimed to assess the genetic diversity and population structure of a Tartary buckwheat germplasm collection from Yunnan Province and adjacent regions to establish a core collection. Whole-genome resequencing and phenotyping of four key agronomic traits were performed on 313 Tartary buckwheat accessions. Population genetic structure, differentiation, and diversity parameters were analyzed using single-nucleotide polymorphism (SNP) data. We obtained 3,433,676 high-quality SNP markers. The 313 accessions were classified into four ancestral populations and three phylogenetic groups, which revealed the complex patterns of genetic differentiation and gene flow. Phenotypic traits exhibited high genetic diversity, with wide variation ranges in key agronomic traits such as plant height, stem diameter, and branching characteristics, highlighting the richness of the germplasm resources. By integrating the phenotypic and SNP data, we constructed a core collection (Core-Merge) comprising 105 accessions (33.55% of the original collection). Core-Merge showed no significant differences in phenotypic traits compared to the original collection, exhibited a similar distribution in principal coordinate analysis, and demonstrated low kinship among individuals. The collection established in this study, Core-Merge, captures the maximum phenotypic and genotypic variability present in the original germplasm. The core germplasm collection provides a valuable foundation for the efficient conservation of, genetic research on, and molecular breeding of Tartary buckwheat. Full article
(This article belongs to the Section Plant Genetic Resources)
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13 pages, 2433 KB  
Article
Development of a Two-Set Multiplex PCR System for Rapid Discrimination of Seven Commercially Important Cuttlefish Species Using COI-Derived SNP Markers
by Chun Mae Dong, Mi-Nan Lee, Hee Jeong Park, Hyo Sun Jung, Eun Soo Noh, In Joon Hwang, Jung-Ha Kang and Eun-Mi Kim
Fishes 2026, 11(4), 226; https://doi.org/10.3390/fishes11040226 - 12 Apr 2026
Viewed by 284
Abstract
Reliable identification of seafood species is critical for fisheries management and product authentication, especially when morphological characteristics are lost during processing. In this study, a multiplex PCR system was developed to distinguish seven cuttlefish species (six Sepia spp. and Sepiella inermis) commercially [...] Read more.
Reliable identification of seafood species is critical for fisheries management and product authentication, especially when morphological characteristics are lost during processing. In this study, a multiplex PCR system was developed to distinguish seven cuttlefish species (six Sepia spp. and Sepiella inermis) commercially distributed in the Korean seafood market. Species identity was first confirmed by amplifying a mitochondrial cytochrome c oxidase subunit I (COI) fragment (~658 bp) using universal primers (LCO1490/HCO2198), showing 99–100% sequence similarity to corresponding GenBank reference sequences. Analysis of genetic variation based on a 530 bp aligned region demonstrated complete interspecific differentiation without shared haplotypes among species. The number of haplotypes per species ranged from 5 to 21, with haplotype diversity values between 0.667 and 1.000. An extended COI fragment (~1200 bp) was further analyzed to identify diagnostic interspecific variation for marker development. Seven diagnostic single-nucleotide polymorphism (SNP) sites were identified and used to design species-specific forward primers with diagnostic nucleotides positioned at the 3′ termini. Distinct amplicons (220–1099 bp) were generated and clearly resolved by agarose gel electrophoresis. Because simultaneous amplification of all seven primer pairs reduced amplification efficiency, the assay was divided into two multiplex sets. Under optimized conditions (56 °C), each species produced a single expected band without cross-amplification. This multiplex PCR system provides a rapid and sequencing-free approach for reliable species discrimination and can be effectively applied to fisheries monitoring and seafood authentication in commercial supply chains. Full article
(This article belongs to the Special Issue Conservation and Population Genetics of Fishes)
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23 pages, 1306 KB  
Review
DNA Mixture Deconvolution: A Four-Strategy Framework from Physical Separation to Database Searching
by Qiang Zhu, Zhigang Mao and Ji Zhang
Genes 2026, 17(4), 434; https://doi.org/10.3390/genes17040434 - 9 Apr 2026
Viewed by 416
Abstract
DNA mixture interpretation remains one of the most technically demanding challenges in forensic genetics. While probabilistic genotyping (PG) systems have substantially advanced likelihood ratio (LR) evaluation, comparatively less attention has been devoted to the systematic reconstruction of contributor genotypes, particularly in no-suspect and [...] Read more.
DNA mixture interpretation remains one of the most technically demanding challenges in forensic genetics. While probabilistic genotyping (PG) systems have substantially advanced likelihood ratio (LR) evaluation, comparatively less attention has been devoted to the systematic reconstruction of contributor genotypes, particularly in no-suspect and database-search contexts. This review synthesizes recent developments in DNA mixture deconvolution through a four-strategy framework: (i) physical and biological separation, (ii) high-information genetic markers, (iii) continuous probabilistic algorithms, and (iv) integration with database searching infrastructures. Upstream approaches, including single-cell isolation and sequencing, reduce mixture complexity at the molecular level. Marker innovations such as microhaplotypes, MiniHaps and DIP-STRs increase per-locus information content and enhance resistance to degradation. Downstream probabilistic models—extended from STRs to SNPs and microhaplotypes—leverage quantitative signal data to infer contributor genotypes, with recent advances in Hamiltonian Monte Carlo, variational inference, and deep learning improving inferential stability and reconstruction accuracy. Importantly, genotype deconvolution and LR evaluation represent mathematically distinct objectives, requiring different validation metrics and potentially separate architectural optimization. The convergence of molecular innovation, algorithmic refinement, and LR-based database searching is progressively transforming mixture interpretation from a purely evidential assessment into an integrated investigative framework. Future progress will depend on standardized marker panels, deconvolution-specific performance metrics, and scalable LR-enabled database infrastructures. Full article
(This article belongs to the Special Issue Advances in Forensic Genetics and DNA)
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21 pages, 17811 KB  
Article
Genome-Wide Association Studies Using Multiple Models Reveal the Genetic Basis of Plant Architecture-Related Traits in Maize
by Beibei Wang, Penghao Wu, Ruotong Wu, Xinru Xie, Zilong Ren, Kaixiang Wang and Jiaojiao Ren
Agronomy 2026, 16(7), 761; https://doi.org/10.3390/agronomy16070761 - 5 Apr 2026
Viewed by 486
Abstract
Plant architecture-related traits are key agronomic traits affecting crop growth and yield. To unravel the genetic architecture of plant height (PH), ear height (EH), tassel length (TL), and tassel primary branch number (TPBN), 379 DH lines derived from 21 maize hybrids were used [...] Read more.
Plant architecture-related traits are key agronomic traits affecting crop growth and yield. To unravel the genetic architecture of plant height (PH), ear height (EH), tassel length (TL), and tassel primary branch number (TPBN), 379 DH lines derived from 21 maize hybrids were used for genome-wide association study (GWAS) and genomic selection (GS) analyses. Although plant architecture-related traits were significantly influenced by genotype and genotype-by-environment interactions, moderate to high broad-sense heritability was observed for PH (81.3%), EH (79.6%), TL (86.4%), and TPBN (82.5%). Using six different models for GWAS, seven unique SNPs on chromosomes 1, 2, and 3 were identified for PH, 92 unique SNPs located on chromosomes 1 to 9 were identified for EH, three unique SNPs on chromosome 6 were detected for TL, and 18 unique SNPs located on chromosomes 1, 4, 5, 8, and 10 were identified for TPBN at the p-value threshold of 7.42 × 10−6. A few hotspot genomic regions conferring plant architecture-related traits were identified, located in bins 2.07, 4.07, 8.03, 6.01, and 10.00. A total of 144 putative candidate genes were identified, which were enriched in endocytosis and lipid biosynthetic process, electron carrier activity, chloroplast stroma, and plastid stroma. The prediction accuracy evaluated through 5-fold cross-validation was 0.44 for PH, 0.43 for EH, 0.31 for TL, and 0.30 for TPBN. When the training population size (TPS) reached 60–70% or marker density (MD) reached 3000, the prediction accuracy tends to stabilize, indicating that the optimum size of TPS and MD were 60–70% and 3000 for GS, respectively. The highest prediction accuracy evaluated by using 30–5000 significant SNPs corresponding to the lowest p-value was 0.70 for PH, 0.85 for EH, 0.58 for TL, and 0.75 for TPBN, with an increase in accuracy of 59.1% to 150.0%. These results demonstrate that integrating GS with a subset of highly significant SNPs can substantially enhance prediction efficiency, thereby facilitating the selection of superior genotypes and accelerating the breeding of maize varieties with optimized plant architecture. This study has further elucidated the genetic basis of maize architecture-related traits and provided valuable information on how to implement GS to breed novel maize varieties with optimized plant types. Full article
(This article belongs to the Section Crop Breeding and Genetics)
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Article
Genetic Variations and Epistatic Interactions for Agronomic and Yield Traits in Winter Wheat Population Derived from ‘TAM 204’ and ‘Iba’ Cultivars
by Yahya Rauf, Jorge Luis Valenzuela-Antelo, Mehmet Dogan, Chenggen Chu, Shannon A. Baker, Jason A. Baker, Daniel Hathcoat, Geraldine Opena, Qingwu Xue, Jackie C. Rudd, Amir M. H. Ibrahim, Junli Zhang and Shuyu Liu
Agronomy 2026, 16(7), 755; https://doi.org/10.3390/agronomy16070755 - 2 Apr 2026
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Abstract
Background: Improving grain yield in wheat remains a top priority, requiring integrated breeding and genetic strategies. This complexity poses a major challenge, driven by quantitative polygenic inheritance, environmental influence, and intricate genetic interactions. We investigated genetic factors and their interactions for agronomic and [...] Read more.
Background: Improving grain yield in wheat remains a top priority, requiring integrated breeding and genetic strategies. This complexity poses a major challenge, driven by quantitative polygenic inheritance, environmental influence, and intricate genetic interactions. We investigated genetic factors and their interactions for agronomic and yield traits in two high-yielding winter wheat cultivars adapted to the US Southern Great Plains. Methods: A bi-parental mapping population consisting of 221 F7 recombinant inbred lines (RIL) derived from ‘TAM 204’ and ‘Iba’ was evaluated for three years in 11 Texas environments. Both parents and RIL population were genotyped on Illumina NovaSeq 6000 and sequences were aligned to IWGSC RefSeq v1.0 using Bowtie2 for SNP calling. For QTL analyses, each trait was analyzed by individual environment, across multiple environments and mega-environments. Results: A total of 86 QTL were mapped for five traits and among them 32 were consistent in more than one environment or analysis. Among consistent QTL, four were pleiotropic to more than one agronomic or yield traits mapped on chromosomes 2B (57.18, 59.47 Mb) and 2D (29.34, 40.64 Mb). The consistent QTL on chromosome 2D (29.34 Mb) was pleiotropic to GYLD, DTH, TW, TKW and explained maximum phenotypic variation for all traits, representing photoperiod gene (Ppd-D1). Another QTL on chromosome 2D (40.64 Mb) was pleiotropic to GYLD and TW and based on the physical position comparisons it likely reflects a unique locus in Iba. The pleiotropic consistent QTL Qgyld.tamu.2B.59 from TAM 204 represents Ppd-B1 gene. Moreover, it is more likely that Qdth.tamu.5B.575 represents the Vrn-B1 gene in Iba. A total of 23 digenic epistatic interactions involved consistent QTL for all traits. Amongst these, epistatic interactions between the consistent QTL on 2B (57.18 Mb) and 2D (29.34 Mb) were observed for GYLD, DTH and TKW. Conclusions: Our findings revealed key allelic diversity and interaction effects in elite wheat cultivars, paving the way for marker development for identified pleiotropic loci and implementation in marker-assisted selection and recombination breeding. Full article
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