Characterization, comparison, and phylogenetic analyses of chloroplast genomes of Euphorbia species.

The genus Euphorbia (Euphorbiaceae) has near-cosmopolitan distribution and serves as a significant resource for both ornamental and medicinal purposes. Despite its economic importance, Euphorbia's taxonomy has long been challenged by the intricate nature of morphological traits exhibiting high levels of convergence. While molecular markers are essential for phylogenetic studies, their availability for Euphorbia has been limited. To address this gap, we conducted comparative analyses focusing on the chloroplast (CP) genomes of nine Euphorbia species, incorporating three newly sequenced and annotated accessions. In addition, phylogenetic informativeness and nucleotide diversity were computed to identify candidate markers for phylogenetic analyses among closely related taxa in the genus. Our investigation revealed relatively conserved sizes and structures of CP genomes across the studied species, with notable interspecific variations observed primarily in non-coding regions and IR/SC borders. By leveraging phylogenetic informativeness and nucleotide diversity, we identified rpoB gene as the optimal candidate for species delimitation and shallow-level phylogenetic inference within the genus. Through this comprehensive analysis of CP genomes across multiple taxa, our study sheds light on the evolutionary dynamics and taxonomic intricacies of Euphorbia, offering valuable insights into its CP genome evolution and taxonomy.

Unveiling a potential threat to forest ecosystems: molecular diagnosis of Alliaria petiolata, a newly introduced alien plant in Korea.

Identifying stages of a species invasion in a new habitat (i.e., colonization, establishment, and landscape spread) and their primary determinants in biological invasion warrants attention, as it provides vital insights for preventing non-native species from becoming pervasive invaders. However, delineating invasion stages and their associated factors can pose significant challenges due to the ambiguous distinctions between these stages. Alliaria petiolata, one of the most noxious weeds in woodland habitats, has recently been introduced to Korea and observed in a few distant locations. Although the plant's spread has been relatively slow thus far, rapid spread is highly likely in the future, given the high invasive potential reported elsewhere. We indirectly diagnose the current status of A. petiolata invasion in Korea through the assessment of genetic diversity and phylogenetic inferences using genome-wide molecular markers and cytological data. We analyzed 86 individual samples collected from two native and six introduced populations, employing 1,172 SNPs. Our analysis estimated within- and among-population genetic diversity and included two clustering analyses. Furthermore, we investigated potential gene flow and reticulation events among the sampled populations. Our data unraveled that Korean garlic mustard exhibits a hexaploid ploidy level with two distinct chromosome numbers, 2n = 36 and 42. The extent of genetic diversity measured in Korean populations was comparable to that of native populations. Using genome-wide SNP data, we identified three distinct clusters with minor gene flow, while failing to detect indications of reticulation among Korean populations. Based on the multifaceted analyses, our study provides valuable insights into the colonization process and stressed the importance of closely monitoring A. petiolata populations in Korea.

The complete chloroplast genome of Euphorbia prostrata (Euphorbiaceae).

Euphorbia prostrata Aiton 1789, an annual herb, is native to tropical and subtropical Americas and was introduced into many parts of the Old-World including Korea. We determined the genomic characteristics of the complete chloroplast genome of E. prostrata with a de novo assembly strategy. The complete chloroplast genome was 162,858 bp long, and harbored 86 protein coding, 37 tRNA and 8 rRNA. The genome showed the typical quadripartite structure consisting of a large single copy (LSC) (90,580 bp), a small single copy (SSC) (18,570 bp) and a pair of inverted repeats (IRs) (26,854 bp). The phylogenetic tree inferred from chloroplast genomes of 25 taxa, which belong to Euphorbiaceae indicated that the Euphorbia is a monophyletic group and E. prostrata is closely related with E. humifusa, E. thymifolia and E. hirta, forming a well-supported clade. Our findings can be valuable for understanding the phylogenetic relationships and the evolution in Euphorbiaceae and will provide basic information for future studies on E. prostrata from genomic perspective.

Complete chloroplast genome sequence of Eranthis byunsanensis B.Y. Sun (Ranunculaceae), an endemic species in Korea.

The Korean endemic Eranthis byunsanensis B.Y. Sun, 1993 (Ranunculaceae) is a rare plant distributed in the southwestern part of the Korean Peninsula. The complete chloroplast (cp) genome of E. byunsanensis was sequenced by next-generation sequencing (NGS) using an Illumina HiSeq X platform. The cp genome of E. byunsanensis is 160,324 bp in length with 37.9% GC content. It showed a typical quadripartite structure consisting of a pair of inverted repeats (IRs; 28,356 bp), a large single-copy region (LSC; 87,671 bp), and a small single-copy region (SSC; 15,941 bp). The cp genome comprises 130 genes including 85 protein-coding genes (PCGs), 37 tRNA genes, and eight rRNA genes. The molecular phylogenetic analysis indicates that E. byunsanensis is closely related to Eranthis stellata, both of which belong to the genus Eranthis.

Genetic differentiation that is exceptionally high and unexpectedly sensitive to geographic distance in the absence of gene flow: Insights from the genus Eranthis in East Asian regions.

Genetic differentiation between populations is determined by various factors, including gene flow, selection, mutation, and genetic drift. Among these, gene flow is known to counter genetic differentiation. The genus Eranthis, an early flowering perennial herb, can serve as a good model to study genetic differentiation and gene flow due to its easily detectable population characteristics and known reproductive strategies, which can be associated with gene flow patterns. Eranthis populations are typically small and geographically separated from the others. Moreover, previous studies and our own observations suggest that seed and pollen dispersal between Eranthis populations is highly unlikely and therefore, currently, gene flow may not be probable in this genus. Based on these premises, we hypothesized that the genetic differentiation between the Eranthis populations would be significant, and that the genetic differentiation would not sensitively reflect geographic distance in the absence of gene flow. To test these hypotheses, genetic differentiation, genetic distance, isolation by distance, historical gene flow, and bottlenecks were analyzed in four species of this genus. Genetic differentiation was significantly high, and in many cases, extremely high. Moreover, genetic differentiation and geographic distance were positively correlated in most cases. We provide possible explanations for these observations. First, we suggest that the combination of the marker type used in our study (chloroplast microsatellites), genetic drift, and possibly selection might have resulted in the extremely high genetic differentiation observed herein. Additionally, we provide the possibility that genetic distance reflects geographic distance through historical gene flow, or adaptation in the absence of historical gene flow. Nevertheless, our explanations can be more rigorously examined and further refined through additional observations and various population genetic analyses. In particular, we suggest that other accessible populations of the genus Eranthis should be included in future studies to better characterize the intriguing population dynamics of this genus.

Impatienshambaeksanensis (Balsaminaceae), a new species from South Korea.

A new species Impatienshambaeksanensis from Gangwon-do, South Korea, is described and illustrated, based on its morphology and distribution. I.hambaeksanensis is different from I.furcillata, another similar Impatiens species in South Korea, in some ways: I.hambaeksanensis possesses a serrate leaf margin with flat tooth tip, while I.furcillata possesses a crenate leaf margin with erect tooth tip; it has an erect inflorescence, while I.furcillata has a pendulous inflorescence; it has a smaller flower which is 2-2.6 cm long, while I.furcillata has a flower of 2.3-3.2 cm; the flower is white or pinkish-white with yellowish and purplish spots, while I.furcillata has a white lower sepal and pinkish-white lateral united petals with yellowish spots; the distal part of the lower sepal is mostly not coiled or rarely 1-coiled, while that of I.furcillata is never coiled; the spur tip is expanded, round and slightly biparted, while that of I.furcillata is expanded, ellipsoidal and clearly biparted. A taxonomic description, a holotype and photos of morphological characteristics of the new species are provided. A table which includes the morphological comparison and a geographical distribution map are presented as well.

Astilbe uljinensis (Saxifragaceae), a new species from South Korea.

A new species Astilbe uljinensis B.U.Oh & H.J.Choi is described from Gangwon-do and Kyeongsangbuk-do in South Korea based on its morphological characteristics and distributional pattern. A. uljinensis is easily distinguished not only from three other Astilbe species in South Korea, but all other species in the genus by possessing a green young rachis, dense long whitish glandular hairs on the young rachis, dense long brownish glandular hairs on the mature inflorescence, and a slightly undulated margin of leaf epidermal cells. Specific comparisons of morphological features such as the type of the trichome, the shape of the leaf epidermis cell, and the color of the young rachis that differentiate A. uljinensis from Astilbe chinensis, another South Korean Astilbe species, are provided.

The speciation history of northern- and southern-sourced Eranthis (Ranunculaceae) species on the Korean peninsula and surrounding areas.

The temporal and spatial origins and evolution of the genus Eranthis have not been previously studied. We investigated the speciation and establishment histories of four Eranthis species: Eranthis byunsanensis, E. pungdoensis, E. stellata, and E. pinnatifida. The sampling localities were Korea, Japan, Jilin in China, and the area near Vladivostok in Primorskiy, Russia. We used 12 chloroplast microsatellite loci (n = 935 individuals) and two chloroplast noncoding regions (rpl16 intron, petL-psbE intergenic spacer; n = 33 individuals). The genetic diversity, genetic structure, phylogenetic relationships of the four species were analyzed, and their ancestral areas were reconstructed. The high genetic diversity of the Jeju island population of E. byunsanensis and Russian populations of E. stellata indicated these species' northward and southward dispersal, respectively. The genetic structure analyses suggest that the populations in these four species have limited geographical structure, except for the Chinese E. stellata population (SCP). The phylogenetic analyses suggest that E. byunsanensis and E. pinnatifida are sister species and that Chinese SCP may not belong to E. stellata. The ancestral area reconstruction revealed that the most recent common ancestor of the four species existed in the current Chinese habitat of E. stellata. This study shows that E. byunsanensis and E. pinnatifida originated from a southern Eranthis species and speciated into their current forms near Jeju island and near western regions of Japan, respectively, during the Miocene. E. stellata may have dispersed southward on and near the Korean peninsula, though its specific origin remains unclear. Interestingly, the Chinese E. stellata population SCP suggests that the Chinese population might be most ancient among all the four Eranthis species. E. pungdoensis may have allopatrically speciated from E. byunsanensis during the Holocene. The Korean peninsula and the surrounding areas can be considered interesting regions which provide the opportunity to observe both northern- and southern-sourced Eranthis species.

Development and characterization of 24 chloroplast microsatellite markers for two species of Eranthis (Ranunculaceae).

Chloroplast microsatellites for two Korean endemic species, Eranthis byunsanensis and E. pungdoensis (Ranunculaceae), were isolated to address the questions of their distributional patterns and evolutionary relationships, using next-generation sequencing. Twenty-four polymorphic chloroplast microsatellite markers for these two species were developed, and then characterized in 65 individuals (55 individuals of E. byunsanensis and 10 individuals of E. pungdoensis). The number of alleles per locus ranged from 2 to 9; the average number of alleles across all the loci scored 4.792. The unbiased diversity per locus ranged from 0.089 to 0.880; the unbiased diversity averaged over all the loci was 0.646. The developed markers were successfully amplified for three congeneric species, E. stellata, E. pinnatifida, and E. longistipitata. The markers developed in this study can provide a valuable and important tool for understanding genetic variations, population structures, evolutionary histories and phylogeography of E. byunsanensis, E. pungdoensis, and related species.

Radiotherapy and cellular infiltration of tumor nests.

There are several opinions which deny the therapeutic effects of preoperative irradiation. However, we have obtained favorable results in treating advanced breast and stomach cancers with preoperative irradiation using a less-fractionated irradiation method with a large dose instead of the conventional fractionated one. We adapted this kind of radiation method to improve resectability and to enhance the antigenicity of the cancer cells so as to inhibit the suppression of the immune reaction in the host. After preoperative irradiation, we examined the resected specimen histopathologically. Remarkable cellular infiltrations, such as neutrocytes, lymphocytes, macrophages and plasma cells of the tumor nests were observed; and these cellular infiltrations, after preoperative irradiation, were more remarkable than the cases without radiotherapy or with conventional fractionated radiotherapy. These cellular infiltrations suggest participation of immunoreaction and we are convinced that this kind of preoperative irradiation could enhance positive immunoreactions. The indications are that this kind of preoperative irradiation of advanced breast and stomach cancer significantly improve the cancer's resectability and curability.