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"Won, Hari"
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Exploration of the genetic diversity of Avena Fatua L. (wild oat) through genotyping-by-sequencing and SDS-PAGE
by
Seo, Hyemin
,
Lee, Kyung Jun
,
Won, Hari
in
Agricultural production
,
Agricultural research
,
Agriculture
2025
Background
The consumption of oats has rapidly increased due to their exceptional nutritional value. However, concerns over genetic erosion have emerged as oat breeding programs rely on a highly limited genetic pool. This study aimed to expand the genetic diversity pool of oats by collecting wild oat (
Avena fatua
L.) populations in South Korea and assessing their genetic diversity and seed storage protein patterns.
Results
A total of 237
A. fatua
individuals were collected in 2022 from eight regions in the southwestern coastal areas of South Korea. Genetic diversity and seed storage protein patterns were analyzed using genotyping-by-sequencing (GBS) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The GBS analysis identified 20,836 single-nucleotide polymorphisms (SNPs). An analysis of molecular variance (AMOVA) based on regional populations revealed that 40.9% of the genetic variation was attributed to differences among populations, while 59.1% was within populations, indicating high genetic differentiation within regional populations. Subsequent population structure analysis and discriminant analysis of principal components (DAPC) both stated the formation of two distinct genetic groups, with an AMOVA value of 70.9% between the groups, suggesting a high level of genetic variation. Pairwise
F
ST
analysis was conducted to compare the genetic differentiation between two populations, revealing that Jindo and Jangheung exhibited the highest level of genetic differentiation (
F
ST
= 0.795) among the geographic groups. Seed storage proteins were analyzed using SDS-PAGE, and the patterns were grouped using k-means clustering. A comparison between the groups based on protein patterns and those based on genetic variation revealed no significant correlation.
Conclusion
This study provides data on the genetic diversity of
A. fatua
, a wild relative of cultivated oats, aimed at expanding the genetic pool of oats for future breeding programs. These findings are expected to be a foundational resource for oat breeding and genetic improvement efforts.
Journal Article
Microbiome of wild oysters: assessing diversity and detecting potential pathogens using a metabarcoding approach
2025
Background
The Pacific oyster (
Crassostrea gigas
) and the hooded oyster (
Saccostrea cucullate
) are ecologically and economically important species in the northwestern Pacific Ocean. However, sustainable oyster farming faces challenges from pathogenic diseases and environmental changes. Understanding microbial diversity in oysters is essential for managing pathogens, maintaining healthy microbial communities, and addressing microbial imbalances. This study aimed to investigate the diversity and composition of bacterial and protist communities in wild oysters from South Korea (
C. gigas
), Taiwan, and the Philippines (
S. cucullata
) using a metabarcoding approach.
Results
Gill tissue and environmental DNA (eDNA) samples were analyzed to assess microbial community structure across species and regions. Bacterial richness significantly exceeded protist richness in all samples. Bacterial diversity was negatively correlated with sea surface temperature and positively correlated with latitude, indicating that temperature is a key driver of regional variation in bacterial community composition. Several potentially pathogenic protist and bacterial taxa were detected, including
Perkinsus marinus
,
Bonamia ostreae
,
Haplosporidium costale
, and
Vibrio bathopelagicus
. Notably,
P. marinus
and
B. ostreae
were identified in
S. cucullata
from Taiwan, while
H. costale
was detected in
C. gigas
from South Korea. Most pathogens occurred at low infection intensities and without clinical signs of disease. Challenges in detecting protist diversity due to sequencing depth and host-derived nontarget amplification were also noted, highlighting the importance of using protist-specific primers.
Conclusions
This study demonstrates the utility of metabarcoding for characterizing microbial communities and detecting pathogens in two oyster species,
C. gigas
and
S. cucullate
, across geographically distinct regions. Bacterial richness was influenced by environmental gradients such as sea surface temperature and latitude, while several key protist pathogens (
P. marinus
,
B. ostreae
,
H. costale
) were detected at low infection intensities in asymptomatic oysters. Limitations in protist detection due to sequencing depth and host amplification highlight the need for optimized primers and complementary approaches such as multiomics and microscopy. These findings provide a foundation for understanding host–microbe–environment interactions. Specifically, our results can inform targeted monitoring programs for the early detection of pathogens and guide selective breeding initiatives for disease-resistant oyster strains, thereby enhancing the long-term resilience of oyster aquaculture under changing environmental conditions.
Journal Article
Widespread admixture blurs population structure and confounds Lake Trout (Salvelinus namaycush) conservation even in the genomic era
by
Boguski, David A.
,
Fraser, Dylan J.
,
Michaelides, Sozos
in
631/158/672
,
631/208/212/2303
,
Animals
2024
Intraspecific variation is important for species’ long-term persistence in changing environments. Conservation strategies targeting intraspecific variation often rely on the identification of management or policy units below the species level based on biological differences among populations. To identify management units, this paper examines intraspecific divergence of Lake Trout (
Salvelinus namaycush)
in Great Slave Lake (GSL), Canada, using low-coverage whole-genome sequencing data. Specifically, we evaluate genetic differentiation and assess the relationship with morphological, mitochondrial, and putatively adaptive divergence. We show that at least three and up to five genetically distinct Lake Trout populations co-occur in GSL and exhibit differences in spatial distribution and body size, with signatures of selection. However, admixture was widespread (60% of the fish) and population structure was shallow (average
F
ST
= 0.022). These findings highlight that, even in the era of whole genome sequencing, identifying discrete management units to implement conservation efforts and policy can remain challenging in systems where gene flow among genetically and ecologically distinct populations is ubiquitous. To give more recognition to this complexity, management efforts should also focus on the area where adaptive variation is evident, and evolutionary acts are at play, to better protect species’ resilience and adaptive potential in some natural systems.
Journal Article
Differential patterns of diversity at neutral and adaptive loci in endangered Rhodeus pseudosericeus populations
2021
Given the fact that threatened species are often composed of isolated small populations, spatial continuity or demography of the populations may be major factors that have shaped the species’ genetic diversity. Thus, neutral loci have been the most commonly-used markers in conservation genetics. However, the populations under the influence of different environmental factors may have evolved in response to different selective pressures, which cannot be fully reflected in neutral genetic variation.
Rhodeus pseudosericeus
, a bitterling species (Acheilognathidae; Cypriniformes) endemic to the Korean Peninsula, are only found in some limited areas of three rivers, Daecheon, Han and Muhan, that flow into the west coast. Here, we genotyped 24 microsatellite loci and two loci (DAB1 and DAB3) of MHC class II peptide-binding β1 domain for 222 individuals collected from seven populations. Our microsatellite analysis revealed distinctive differentiation between the populations of Daecheon and Muhan Rivers and the Han River populations, and populations were structured into two subgroups within the Han River. Apparent positive selection signatures were found in the peptide-binding residues (PBRs) of the MHC loci. The allelic distribution of MHC showed a degree of differentiation between the populations of Daecheon and Muhan Rivers and the Han River populations, partially similar to the results obtained for microsatellites, however showed rather complex patterns among populations in the Han River. Considering the apparent differences in the distribution of supertypes obtained based on the physicochemical differences induced by the polymorphisms of these PBRs, the differentiation in DAB1 between the two regional groups may result in the differences in immune function. No differentiation between these two regions was observed in the supertyping of DAB3, probably indicating that only DAB1 was associated with the response to locally specialized antigenic peptides.
Journal Article
The first Chromosomal-level genome assembly of Sageretia thea using Nanopore long reads and Pore-C technology
by
Lee, Kyung Jun
,
Won, Hari
,
Park, Jong-Soo
in
631/208/212/748
,
631/449/2669
,
Chromatin remodeling
2024
Sageretia thea
, a notable species within the mock buckthorn genus, is recognized for its intriguing biogeographical distribution and diverse medicinal properties. Despite this significance, genomic studies on
S. thea
are still in the nascent stages. We present the first chromosome-level genome assembly of
S. thea
that was generated using a combination of Oxford Nanopore long-read and Illumina short-read sequencing technologies complemented by Pore-C chromatin conformation capture. The genome assembly had a size of 197.8 Mb with 12 chromosomal scaffolds and a scaffold N50 length of 15.9 Mb. A total of 25,434 protein-coding genes were identified and functionally annotated, and the gene model indicated 96.5% complete eukaryotic BUSCOs. Additionally, orthologous gene profiling and synteny analysis were performed to elucidate the evolutionary relationships within the Rhamnaceae family and Rosales. This high-quality chromosomal genome is the first genomic view of
S. thea
, which will serve as the basis for future studies on its biological and medicinal properties, and evolutionary history.
Journal Article
Evidence of an ancient connectivity and biogeodispersal of a bitterling species, Rhodeus notatus, across the Korean Peninsula
2020
The modern-day distribution of freshwater fishes throughout multiple rivers is likely the result of past migration during times when currently separate drainages were once connected. Here, we used mitochondrial and microsatellite analyses for 248 individuals of
Rhodeus notatus
collected from seven different rivers to obtain better understand historical gene flow of freshwater fish on the Korean Peninsula. Based on our phylogenetic analyses, this Korean species originated through the paleo-Yellow River from China and first colonized near the west coast. These genetic data also provided evidence of estuary coalescences among the rivers flowing to the west and southwest coast on well-developed continental shelf. In addition, the pattern of population structure revealed the biogeodispersal route from the west coast to the south coast. It could be inferred that massive migration was not involved in the formation of southern populations, since the signature of historical genetic drift was clearly observed. Our study is the first genetic attempt to confirm hypotheses describing the migration of freshwater species towards the end of East Asia, which have previously been developed using only geological reasoning.
Journal Article
Polymorphism of MHC class IIB in an acheilognathid species, Rhodeus sinensis shaped by historical selection and recombination
2019
Background Rhodeus sinensis is a bitterling species occurring throughout the numerous freshwater systems on the East Asia. Here, we analyzed the diversity of the MHC class IIB (DAB) genes from this species, which may offer meaningful insights into evolutionary processes in this species as well as other bitterlings. Results Using cDNA and gDNA samples from 50 individuals, we discovered classical 140 allelic sequences that could be allocated into either DAB1 (Rhsi-DAB1) or DAB3 (Rhsi-DAB3). DAB sequences completely lacking the intron, but identical or similar to Rhsi-DAB1, were also discovered from our gDNA samples, and this intron loss likely originated from the retrotransposition events of processed mDNA. The β1 domain was the most polymorphic in both Rhsi-DAB1 and -DAB3. Putative peptide biding residues (PBRs) in Rhsi-DAB1, but not in Rhsi-DAB3, exhibited a significant dN/dS, presumably indicating that different selection pressures have acted on those two DABs. Recombination between different alleles seemed to have contributed to the increase of diversity in Rhsi-DABs. Upon phylogenetic analysis, Rhsi-DAB1 and -DAB3 formed independent clusters. Several alleles from other species of Cypriniformes were embedded in the clade of Rhsi-DAB1, whereas Rhsi-DAB3 clustered with alleles from the wider range of taxa (Cyprinodontiformes), indicating that these two Rhsi-DABs have taken different historical paths. Conclusions A great deal of MHC class IIB allelic diversity was found in R. sinensis, and gene duplication, selection and recombination may have contributed to this diversity. Based on our data, it is presumed that such historical processes have commonly or differently acted on the polymorphism of Rhsi-DAB1 and -DAB3.
Journal Article
Methionine consumption by cancer cells drives a progressive upregulation of PD-1 expression in CD4 T cells
2023
Programmed cell death protein 1 (PD-1), expressed on tumor-infiltrating T cells, is a T cell exhaustion marker. The mechanisms underlying PD-1 upregulation in CD4 T cells remain unknown. Here we develop nutrient-deprived media and a conditional knockout female mouse model to study the mechanism underlying PD-1 upregulation. Reduced methionine increases PD-1 expression on CD4 T cells. The genetic ablation of SLC43A2 in cancer cells restores methionine metabolism in CD4 T cells, increasing the intracellular levels of S-adenosylmethionine and yielding H3K79me2. Reduced H3K79me2 due to methionine deprivation downregulates AMPK, upregulates PD-1 expression and impairs antitumor immunity in CD4 T cells. Methionine supplementation restores H3K79 methylation and AMPK expression, lowering PD-1 levels. AMPK-deficient CD4 T cells exhibit increased endoplasmic reticulum stress and
Xbp1s
transcript levels. Our results demonstrate that AMPK is a methionine-dependent regulator of the epigenetic control of PD-1 expression in CD4 T cells, a metabolic checkpoint for CD4 T cell exhaustion.
The deprivation of amino acids in the tumor microenvironment affects T cell survival and activation. Here the authors show that reduced levels of methionine are associated with PD1 upregulation in CD4+ T cells and that methionine supplementation promotes CD4+ T cell dependent anti-tumor immune responses.
Journal Article
Effect of Heat Stress on Seed Protein Composition and Ultrastructure of Protein Storage Vacuoles in the Cotyledonary Parenchyma Cells of Soybean Genotypes That Are Either Tolerant or Sensitive to Elevated Temperatures
by
Krishnan, Hari B.
,
Smith, James R.
,
Kim, Won-Seok
in
Antibodies
,
Cotyledon - chemistry
,
Cotyledon - ultrastructure
2020
High growth temperatures negatively affect soybean (Glycine max (L.) Merr) yields and seed quality. Soybean plants, heat stressed during seed development, produce seed that exhibit wrinkling, discoloration, poor seed germination, and have an increased potential for incidence of pathogen infection and an overall decrease in economic value. Soybean breeders have identified a heat stress tolerant exotic landrace genotype, which has been used in traditional hybridization to generate experimental genotypes, with improved seed yield and heat tolerance. Here, we have investigated the seed protein composition and ultrastructure of cotyledonary parenchyma cells of soybean genotypes that are either susceptible or tolerant to high growth temperatures. Biochemical analyses of seed proteins isolated from heat-tolerant and heat-sensitive genotypes produced under 28/22 °C (control), 36/24 °C (moderate), and 42/26 °C (extreme) day/night temperatures revealed that the accumulation in soybean seeds of lipoxygenase, the β-subunit of β-conglycinin, sucrose binding protein and Bowman-Birk protease inhibitor were negatively impacted by extreme heat stress in both genotypes, but these effects were less pronounced in the heat-tolerant genotype. Western blot analysis showed elevated accumulation of heat shock proteins (HSP70 and HSP17.6) in both lines in response to elevated temperatures during seed fill. Transmission electron microscopy showed that heat stress caused dramatic structural changes in the storage parenchyma cells. Extreme heat stress disrupted the structure and the membrane integrity of protein storage vacuoles, organelles that accumulate seed storage proteins. The detachment of the plasma membrane from the cell wall (plasmolysis) was commonly observed in the cells of the sensitive line. In contrast, these structural changes were less pronounced in the tolerant genotype, even under extreme heat stress, cells, for the most part, retained their structural integrity. The results of our study demonstrate the contrasting effects of heat stress on the seed protein composition and ultrastructural alterations that contribute to the tolerant genotype’s ability to tolerate high temperatures during seed development.
Journal Article
Bowman–Birk Inhibitor Mutants of Soybean Generated by CRISPR-Cas9 Reveal Drastic Reductions in Trypsin and Chymotrypsin Inhibitor Activities
2024
Despite the high quality of soybean protein, raw soybeans and soybean meal cannot be directly included in animal feed mixtures due to the presence of Kunitz (KTi) and Bowman–Birk protease inhibitors (BBis), which reduces animal productivity. Heat treatment can substantially inactivate trypsin and chymotrypsin inhibitors (BBis), but such treatment is energy-intensive, adds expense, and negatively impacts the quality of seed proteins. As an alternative approach, we have employed CRISPR/Cas9 gene editing to create mutations in BBi genes to drastically lower the protease inhibitor content in soybean seed. Agrobacterium-mediated transformation was used to generate several stable transgenic soybean events. These independent CRISPR/Cas9 events were examined in comparison to wild-type plants using Sanger sequencing, proteomic analysis, trypsin/chymotrypsin inhibitor activity assays, and qRT-PCR. Collectively, our results demonstrate the creation of an allelic series of loss-of-function mutations affecting the major BBi gene in soybean. Mutations in two of the highly expressed seed-specific BBi genes lead to substantial reductions in both trypsin and chymotrypsin inhibitor activities.
Journal Article