Genome assembly and microsatellite marker development using Illumina and PacBio sequencing in Persicaria maackiana (Polygonaceae) from Korea.

BACKGROUND: Persicaria maackiana (Regel) is a potential medicinal plant that exerts anti-diabetic effects. However, the lack of genomic information on P. maackiana hinders research at the molecular level. OBJECTIVE: Herein, we aimed to construct a draft genome assembly and obtain comprehensive genomic information on P. maackiana using high-throughput sequencing tools PacBio Sequel II and Illumina. METHODS: Persicaria maackiana samples from three natural populations in Gaecheon, Gichi, and Uiryeong reservoirs in South Korea were used to generate genomic DNA libraries, perform genome de novo assembly, gene ontology analysis, phylogenetic tree analysis, genotyping, and identify microsatellite markers. RESULTS: The assembled P. maackiana genome yielded 32,179 contigs. Assessment of assembly integrity revealed 1503 (93.12%) complete Benchmarking Universal Single-Copy Orthologs. A total of 64,712 protein-coding genes were predicted and annotated successfully in the protein database. In the Kyoto Encyclopedia of Genes and Genomes (KEGG) orthologs, 13,778 genes were annotated into 18 categories. Genes that activated AMPK were identified in the KEGG pathway. A total of 316,992 microsatellite loci were identified, and primers targeting the flanking regions were developed for 292,059 microsatellite loci. Of these, 150 primer sets were randomly selected for amplification, and 30 of these primer sets were identified as polymorphic. These primers amplified 3-9 alleles. The mean observed and expected heterozygosity were 0.189 and 0.593, respectively. Polymorphism information content values of the markers were 0.361-0.754. CONCLUSION: Collectively, our study provides a valuable resource for future comparative genomics, phylogeny, and population studies of P. maackiana.

Genome Assembly and Microsatellite Marker Development Using Illumina and PacBio Sequencing in the Carex pumila (Cyperaceae) from Korea.

This study is the first to report the characterization of Carex pumila genomic information. Assembly of the genome generated a draft of C. pumila based on PacBio Sequel II and Illumina paired-end sequencing, which was assembled from 2941 contigs with an estimated genome size of 0.346 Gb. The estimate of repeats in the genome was 31.0%, and heterozygosity ranged from 0.426 to 0.441%. The integrity evaluation of the assembly revealed 1481 complete benchmarked universal single-copy orthologs (BUSCO) (91.76%), indicating the high quality of the draft assembly. A total of 23,402 protein-coding genes were successfully predicted and annotated in the protein database. UpsetR plots showed that 7481 orthogroups were shared by all species. The phylogenetic tree showed that C. pumila is a close but distant relative of Ananas comosus. C. pumila had greater contraction (3154) than expansion (392). Among the extended gene families, aquaporins have been found to be enriched. Primers for microsatellite markers determined 30 polymorphic markers out of 100. The average number of alleles amplified by these 30 polymorphic markers was 4 to 12, with an average polymorphism information content (PIC) value of 0.660. In conclusion, our study provides a useful resource for comparative genomics, phylogeny, and future population studies of C. pumila.

Community recovery of benthic macroinvertebrates in a stream influenced by mining activity: Importance of microhabitat monitoring.

The decrease in freshwater biodiversity owing to anthropogenic disturbances such as mining activity is a global challenge; hence, there is an urgent need for systematic approaches to continuously monitor such disturbances and/or the recovery of biodiversity in freshwater habitats. The Hwangjicheon Stream is the source of South Korea's longest river and has been subjected to runoff from coal mining. We investigated changes in the diversity of the benthic macroinvertebrate community in various microhabitats, including riffle, run, and pool, to monitor the recovery of biodiversity in the stream following the improvement of a mining water treatment plant in 2019. The dataset comprised 111 samples obtained from four types of microhabitats (riffle, run, pool, and riparian) over a four-year period from 2018 to 2021. The mining-affected sites had lower macroinvertebrate community complexities according to a network analysis, and grouped into the same cluster based on self-organizing map (SOM) analysis. Moreover, 51 taxa selected as indicator species represented each cluster obtained through the SOM analysis. Among them, only Limnodrilus gotoi and Radix auricularia were included as indicator species at the mining-affected sites. However, after 2020, the benthic macroinvertebrate community complexity increased, and some of the microhabitats at the mining-affected sites were included in the same cluster as the reference sites in the SOM analysis, indicating that the recovery of benthic macroinvertebrate communities had initiated in certain microhabitats (e.g., riparian). Further analysis confirmed that the macroinvertebrate community clearly differed according to the survey year, even in different microhabitats at the same sites. This suggests that more acute microhabitat monitoring may be necessary to quickly confirm biodiversity restoration when assessing the degree of the recovery in river biodiversity from anthropogenic disturbances.

Complete Chloroplast Genome Determination of Ranunculus sceleratus from Republic of Korea (Ranunculaceae) and Comparative Chloroplast Genomes of the Members of the Ranunculus Genus.

Ranunculus sceleratus (family: Ranunculaceae) is a medicinally and economically important plant; however, gaps in taxonomic and species identification limit its practical applicability. This study aimed to sequence the chloroplast genome of R. sceleratus from Republic of Korea. Chloroplast sequences were compared and analyzed among Ranunculus species. The chloroplast genome was assembled from Illumina HiSeq 2500 sequencing raw data. The genome was 156,329 bp and had a typical quadripartite structure comprising a small single-copy region, a large single-copy region, and two inverted repeats. Fifty-three simple sequence repeats were identified in the four quadrant structural regions. The region between the ndhC and trnV-UAC genes could be useful as a genetic marker to distinguish between R. sceleratus populations from Republic of Korea and China. The Ranunculus species formed a single lineage. To differentiate between Ranunculus species, we identified 16 hotspot regions and confirmed their potential using specific barcodes based on phylogenetic tree and BLAST-based analyses. The ndhE, ndhF, rpl23, atpF, rps4, and rpoA genes had a high posterior probability of codon sites in positive selection, while the amino acid site varied between Ranunculus species and other genera. Comparison of the Ranunculus genomes provides useful information regarding species identification and evolution that could guide future phylogenetic analyses.

The complete chloroplast genome of Persicaria maackiana (Regel) Nakai ex T. Mori (Polygonaceae) in Korea.

Persicaria maackiana (Regel) Nakai ex T. Mori (1922), a species of the Polygonaceae family, is an annual plant widely distributed in Northeast Asia. We aimed to sequence the complete chloroplast genome of P. maackiana using Illumina HiSeq paired-end sequencing. The chloroplast genome was determined to be 160,635 bp. The complete chloroplast genome contained 129 genes, including 84 protein-coding genes, 37 tRNA, and eight rRNA genes. Phylogenetic analysis of the chloroplast genome sequences of 15 Polygonaceae plants revealed that P. maackiana was most closely related to P. perfoliata. Our findings might be useful for future phylogenetic studies of Polygonaceae.

Micromorphological and cytological comparisons between Youngia japonica and Youngia longiflora using light and scanning electron microscopy.

The genus Youngia Cass. is taxonomically complex and widely distributed in East Asia. Despite the high diversity of species, only two species, Youngia japonica (L.) DC. and Youngia longiflora (Babc. & Stebbins) C.Shih, are currently found in Korea. Among them, Y. longiflora was once recognized as a subspecies of Y. japonica. However, based on chloroplast and nuclear DNA sequences, as well as morphology, Y. longiflora was found to be a distinct species. Here, the two Korean Youngia species were analyzed using scanning electron microscope and light microscope for chromosome number, pollen, cypselae, and leaf micromorphological characters to evaluate if the examined characters are informative for taxonomy. Pollen morphology is described, and the micromorphological characters of Y. longiflora are described for the first time. Cypselae and pollen surface ornamentation, as well as trichome morphology and stomatal size varied between the two species. Y. japonica had longer chromosomes than Y. longiflora, while other chromosomal characters including the localization of the NOR (nucleolar organizer region) were similar. The present micromorphological and cytological results are of great taxonomic value for Youngia species identification in Korea.

The location of multifidus atrophy in patients with a single level, unilateral lumbar radiculopathy.

OBJECTIVE: To identify the correlations between the location of multifidus atrophy and the level of lumbar radiculopathy. METHODS: Thirty-seven patients who had unilateral L4 or L5 radiculopathy were divided into 2 groups; the L4 radiculopathy (L4 RAD) group and the L5 radiculopathy (L5 RAD) group. Bilateral lumbar multifidus muscles at the mid-spinous process level of L4 vertebra (L4 MSP), the mid-spinous process level of L5 vertebra (L5 MSP), and the mid-sacral crest level of S1 vertebra (S1 MSC) were detected in T1 axial magnetic resonance imaging. The total muscle cross-sectional area of multifidus muscles (TMCSA) and the pure muscle cross-sectional area of multifidus muscles (PMCSA) were measured by a computerized analysis program, and the ratio of PMCSA to TMCSA (PMCSA/TMCSA) was calculated. RESULTS: There were no significant differences in TMCSA between the involved and the uninvolved sides in both groups. PMCSA was only significantly smaller at the S1 MSC on the involved side as compared with the uninvolved side in the L5 RAD group. The ratio of PMCSA to TMCSA was the lowest at the L5 MSP on the involved side in the L4 RAD group and at the S1 MSC on the involved side in the L5 RAD group. CONCLUSION: Our findings suggest that the most severe atrophy of multifidus muscle may occur at the mid-spinous process or mid-sacral crest level of the vertebra which is one level below the segmental number of the involved nerve root in patients with a single-level, unilateral lumbar radiculopathy.