Wasabi Gone Wild? Origin and Characterization of the Complete Plastomes of Ulleung Island Wasabi (Eutrema japonicum; Brassicaceae) and Other Cultivars in Korea.

Korean wasabi occurs naturally on the young oceanic, volcanic Ulleung Island off the east coast of the Korean Peninsula. Although the Ulleung Island wasabi is reported as Eutrema japonicum and has been suggested to be morphologically identical to cultivars in Korea, very little is known about its taxonomic identity and relationship with other cultivars. In this study, we sequenced the complete chloroplast DNA sequences of three naturally occurring Ulleung Island wasabi plants and six cultivars ('Daewang', 'Daruma', 'Micado', 'Orochi', 'Green Thumb', and 'Shogun') from continental Korea and determined the taxonomic identity of Korean wasabi on Ulleung Island. The size and organization of the complete chloroplast genomes of the nine accessions were nearly identical to those of previously reported wasabi cultivars. In addition, phylogenetic analysis based on the complete plastomes suggested that Ulleung Island wasabi most likely comprises various wasabi cultivars with three chlorotypes ('Shogun', 'Green Thumb', and a unique Chusan type). Based on the complete plastomes, we identified eight chlorotypes for the major wasabi cultivars and the Ulleung Island wasabi. Two major groups (1-'Mazuma' and 'Daruma', and 2-'Fujidaruma'/'Shimane No. 3'/Ulleung Island wasabi/five cultivars in Korea) were also identified based on mother line genealogical history. Furthermore, different types of variations (mutations, insertions/deletions (indels), mononucleotide repeats, and inversions) in plastomes were identified to distinguish different cultivar lines and five highly divergent hotspots. The nine newly obtained complete plastomes are valuable organelle genomic resources for species identification and infraspecific phylogeographic studies on wild and cultivated wasabi.

Chloroplast Genome Provides Insights into Molecular Evolution and Species Relationship of Fleabanes (Erigeron: Tribe Astereae, Asteraceae) in the Juan Fernández Islands, Chile.

Erigeron represents the third largest genus on the Juan Fernández Islands, with six endemic species, five of which occur exclusively on the younger Alejandro Selkirk Island with one species on both islands. While its continental sister species is unknown, Erigeron on the Juan Fernández Islands appears to be monophyletic and most likely evolved from South American progenitor species. We characterized the complete chloroplast genomes of five Erigeron species, including accessions of E. fernandezia and one each from Alejandro Selkirk and Robinson Crusoe Islands, with the purposes of elucidating molecular evolution and phylogenetic relationships. We found highly conserved chloroplast genomes in size, gene order and contents, and further identified several mutation hotspot regions. In addition, we found two positively selected chloroplast genes (ccsA and ndhF) among species in the islands. The complete plastome sequences confirmed the monophyly of Erigeron in the islands and corroborated previous phylogenetic relationships among species. New findings in the current study include (1) two major lineages, E. turricola-E. luteoviridis and E. fernandezia-E. ingae-E. rupicola, (2) the non-monophyly of E. fernandezia occurring on the two islands, and (3) the non-monophyly of the alpine species E. ingae complex.

Plastome-based backbone phylogeny of East Asian Phedimus (Subgenus Aizoon: Crassulaceae), with special emphasis on Korean endemics.

Although the monophyly of Phedimus has been strongly demonstrated, the species relationships among approximately 20 species of Phedimus have been difficult to determine because of the uniformity of their floral characteristics and extreme variation of their vegetative characters, often accompanied by high polyploid and aneuploid series and diverse habitats. In this study, we assembled 15 complete chloroplast genomes of Phedimus species from East Asia and generated a plastome-based backbone phylogeny of the subgenus Aizoon. As a proxy for nuclear phylogeny, we reconstructed the nuclear ribosomal DNA internal transcribed spacer (nrDNA ITS) phylogeny independently. The 15 plastomes of subg. Aizoon were highly conserved in structure and organization; hence, the complete plastome phylogeny fully resolved the species relationships with strong support. We found that P. aizoon and P. kamtschaticus were polyphyletic and morphologically distinct or ambiguous species, and they most likely evolved from the two species complex. The crown age of subg. Aizoon was estimated to be 27 Ma, suggesting its origin to be in the late Oligocene; however, the major lineages were diversified during the Miocene. The two Korean endemics, P. takesimensis and P. zokuriensis, were inferred to have originated recently during the Pleistocene, whereas the other endemic, P. latiovalifolium, originated in the late Miocene. Several mutation hotspots and seven positively selected chloroplast genes were identified in the subg. Aizoon.

Phylogenomic insight into dysploidy, speciation, and plastome evolution of a small Mediterranean genus Reichardia (Cichorieae; Asteraceae).

Reichardia Roth is a small Mediterranean genus comprising ten homogeneous species with basic chromosome numbers of 7, 8, and 9. To assess the plastid genome evolution and differentiation of Reichardia species, we assembled the complete plastome sequences of seven Reichardia and two Launaea species and conducted various phylogenomic analyses comparatively with nuclear ribosomal DNA ITS sequences. Reichardia and Launaea plastomes were highly conserved in gene content and order, containing 130 genes. Plastid phylogenomic reconstruction strongly suggested that Reichardia was a sister to Launaea, and its common ancestor initially diverged into two major lineages: the first containing species with n = 8 chromosomes exclusively, and the other with n = 9, 8, and 7 chromosomes. Although the ancestral Reichardia karyotype was suggested to most likely be n = 9 from ancestral chromosome number reconstruction, the pattern of descending dysploidy indicated by the phylogenetic trees based on nuclear ribosomal DNA ITS was less evident in the trees based on the plastome. Possible reasons for these findings are discussed.

Genome-Wide Single Nucleotide Polymorphism Analysis Elucidates the Evolution of Prunus takesimensis in Ulleung Island: The Genetic Consequences of Anagenetic Speciation.

Of the two major speciation modes of endemic plants on oceanic islands, cladogenesis and anagenesis, the latter has been recently emphasized as an effective mechanism for increasing plant diversity in isolated, ecologically homogeneous insular settings. As the only flowering cherry occurring on Ulleung Island in the East Sea (concurrently known as Sea of Japan), Prunus takesimensis Nakai has been presumed to be derived through anagenetic speciation on the island. Based on morphological similarities, P. sargentii Rehder distributed in adjacent continental areas and islands has been suggested as a purported continental progenitor. However, the overall genetic complexity and resultant non-monophyly of closely related flowering cherries have hindered the determination of their phylogenetic relationships as well as the establishment of concrete continental progenitors and insular derivative relationships. Based on extensive sampling of wild flowering cherries, including P. takesimensis and P. sargentii from Ulleung Island and its adjacent areas, the current study revealed the origin and evolution of P. takesimensis using multiple molecular markers. The results of phylogenetic reconstruction and population genetic structure analyses based on single nucleotide polymorphisms detected by multiplexed inter-simple sequence repeat genotyping by sequencing (MIG-seq) and complementary cpDNA haplotypes provided evidence for (1) the monophyly of P. takesimensis; (2) clear genetic differentiation between P. takesimensis (insular derivative) and P. sargentii (continental progenitor); (3) uncertain geographic origin of P. takesimensis, but highly likely via single colonization from the source population of P. sargentii in the Korean Peninsula; (4) no significant reduction in genetic diversity in anagenetically derived insular species, i.e., P. takesimensis, compared to its continental progenitor P. sargentii; (5) no strong population genetic structuring or geographical patterns in the insular derivative species; and (6) MIG-seq method as an effective tool to elucidate the complex evolutionary history of plant groups.

Plastid Phylogenomics of Dendroseris (Cichorieae; Asteraceae): Insights Into Structural Organization and Molecular Evolution of an Endemic Lineage From the Juan Fernández Islands.

Dendroseris D. Don comprises 11 species endemic to the Juan Fernández islands in Chile. They demonstrate spectacular and unusual growth forms of rosette trees with extremely variable morphology and occupy wide ecological ranges on the islands. These unique plants are now highly threatened with extinction with very small population sizes, typically consisting of 10 or fewer individuals in wild. Despite morphological and ecological divergence among species of Dendroseris, their monophyly has been supported in previous studies, but with little resolution among subgeneric groups. We assembled seven complete plastome sequences from seven species of Dendroseris, including representatives from three subgenera, and carried out comparative phylogenomic analyses. The plastomes are highly conserved in gene content and order, with size ranging from 152,199 to 152,619 bp and containing 130 genes (87 coding genes, 6 rRNA genes, and 37 tRNA genes). Plastid phylogenomic analyses based on both the complete plastome sequences and 81 concatenated coding genes only show Dendroseris nested within Sonchus sensu lato, and also that inter-subgeneric relationships are fully resolved. Subg. Phoenicoseris is resolved as sister to the remaining species of the genus and a sister relationship between the two subgenera Dendroseris and Rea. Ten mutation hotspots from LSC and SSC regions and variable SSRs are identified as potential chloroplast markers for future phylogenetic and phylogeographic studies of Sonchus and related groups.

Sow Thistle Chloroplast Genomes: Insights into the Plastome Evolution and Relationship of Two Weedy Species, Sonchus asper and Sonchus oleraceus (Asteraceae).

Prickly sow thistle, Sonchus asper (L.) Hill, and common sow thistle, Sonchus oleraceus L., are noxious weeds. Probably originating from the Mediterranean region, they have become widespread species. They share similar morphology and are closely related. However, they differ in their chromosome numbers and the precise relationship between them remains uncertain. Understanding their chloroplast genome structure and evolution is an important initial step toward determining their phylogenetic relationships and analyzing accelerating plant invasion processes on a global scale. We assembled four accessions of chloroplast genomes (two S. asper and two S. oleraceus) by the next generation sequencing approach and conducted comparative genomic analyses. All the chloroplast genomes were highly conserved. Their sizes ranged from 151,808 to 151,849 bp, containing 130 genes including 87 coding genes, 6 rRNA genes, and 37 tRNA genes. Phylogenetic analysis based on the whole chloroplast genome sequences showed that S. asper shares a recent common ancestor with S. oleraceus and suggested its likely involvement in a possible amphidiploid origin of S. oleraceus. In total, 79 simple sequence repeats and highly variable regions were identified as the potential chloroplast markers to determine genetic variation and colonization patterns of Sonchus species.

Evolutionary Comparison of the Chloroplast Genome in the Woody Sonchus Alliance (Asteraceae) on the Canary Islands.

The woody Sonchus alliance consists primarily of woody species of the genus Sonchus (subgenus Dendrosonchus; family Asteraceae). Most members of the alliance are endemic to the oceanic archipelagos in the phytogeographic region of Macaronesia. They display extensive morphological, ecological, and anatomical diversity, likely caused by the diverse habitats on islands and rapid adaptive radiation. As a premier example of adaptive radiation and insular woodiness of species endemic to oceanic islands, the alliance has been the subject of intensive evolutionary studies. While phylogenetic studies suggested that it is monophyletic and its major lineages radiated rapidly early in the evolutionary history of this group, genetic mechanisms of speciation and genomic evolution within the alliance remain to be investigated. We first attempted to address chloroplast (cp) genome evolution by conducting comparative genomic analysis of three representative endemic species (Sonchus acaulis, Sonchus canariensis, and Sonchus webbii) from the Canary Islands. Despite extensive morphological, anatomical, and ecological differences among them, their cp genomes were highly conserved in gene order and content, ranging from 152,071 to 152,194 bp in total length. The number of repeat variations and six highly variable regions were identified as valuable molecular markers. Phylogenetic analysis of 32 species in the family Asteraceae revealed the phylogenetic position of the woody Sonchus alliance within the tribe Cichorieae and the sister relationship between the weedy Sonchus oleraceus and the alliance.

The complete chloroplast genome of Korean Pyrus ussuriensis Maxim. (Rosaceae): providing genetic background of two types of P. ussuriensis.

Pyrus ussuriensis Maxim. is one of the most important pear species cultivated in Asia, grown in northern China, far-east Russia, Korea, and Japan. Here we completed the chloroplast genome of wild P. ussuriensis collected in Bonghwa-gun, Korea, which was 159,986 bp in length consisting of four subregions: 87,947 bp of large single copy (LSC) and 19,255 bp of small single copy (SSC) regions are separated by 26,392 bp of inverted repeat (IR) regions. The genome contained a total of 130 genes including 85 protein-coding genes, eight rRNAs, and 37 tRNAs. The overall GC content was 36.5% and those in the LSC, SSC, and IR regions were 34.2%, 30.4%, and 42.6%, respectively. Phylogenetic analysis of 14 Pyrus chloroplast genomes provided the diverse genetic background for wild P. ussuriensis populations in Korea by confirming that wild P. ussuriensis sequenced in this study contained Pyrus pyrifolia type plastome. It revealed substantial sequence variations up to 121 single nucleotide polymorphisms and 781 insertions and deletions against another wild accession of P. ussuriensis (P. ussuriensis type) collected in Mt. Hambaek, Korea.

Characterization of the complete chloroplast genome of the woody sow-thistle, Sonchus leptocephalus (Asteraceae) endemic to the Canary Islands.

Sonchus leptocephalus belongs to a core member of the woody Sonchus alliance endemic to the Macaronesian Islands in the Atlantic Ocean. The alliance has been subject to intensive investigations of adaptive radiation on oceanic islands. As an attempt to fully understand the patterns and processes of evolution in this group, we determined the complete chloroplast genome of S. leptocephalus endemic to the Canaries. It was 152,406 bp in length, comprising 84,331 bp of large single copy and 18,583 bp of small single copy separated by 24,746 bp of inverted repeats. A total of 130 genes were determined including 87 protein-coding genes, 6 ribosomal RNA, and 37 transfer RNA genes. Phylogenetic analysis confirmed its position within the woody Sonchus alliance.

Multiple Lines of Evidence for Independent Origin of Wild and Cultivated Flowering Cherry (Prunus yedoensis).

As with many other ornamental and cultivated plants that have been under human selection and cultivation for a long time, it has been a major challenge to trace back the complex evolutionary history of flowering cherry, Prunus yedoensis. This challenge has been further amplified by great morphological similarities, little molecular divergence, frequent natural and artificial hybridization, and poor documentation of breeding history among cultivated and wild flowering cherries. The origin and taxonomic distinction between wild P. yedoensis from Jeju Island, Korea, and one of the most popular cultivated flowering cherries, P. × yedoensis "Somei-yoshino" has been a controversy for the past few decades. We sampled many areas extensively, and using four different molecular markers we provided evidence for their independent origin. Wild P. yedoensis in Korea originated from multiple bidirectional hybridization events between two sympatric species, P. spachiana f. ascendens as the maternal species and P. serrulata var. spontanea/P. serrulata var. quelpaertensis as the most probable paternal species. On the contrary, our results supported a single artificial hybrid origin of P. × yedoensis "Somei-yoshino" from cultivated P. spachiana f. ascendens as the maternal species and P. speciosa, a species endemic to Izu Islands, as the paternal species. Based on extensive sampling, we provided strong evidence that wild and cultivated P. yedoensis are distinct taxonomic entities that have originated from different evolutionary processes. A potential for the development of new cultivars from wild P. yedoensis and conservation of diverse germplasms in situ insular setting and ex situ should be explored in the future.

Phylogeography and Ecological Niche Modeling Reveal Reduced Genetic Diversity and Colonization Patterns of Skunk Cabbage (Symplocarpus foetidus; Araceae) From Glacial Refugia in Eastern North America.

Alternating glacial and interglacial periods during the Quaternary have dramatically affected the distribution and population genetic structure of plant and animal species throughout the northern hemisphere. Surprisingly, little is known about the post-glacial recolonization history of wetland herbaceous perennials that are widely distributed in the understory of deciduous or mixed deciduous-evergreen forests in eastern North America. In this study, we investigated infraspecific variation among 32 populations of skunk cabbage, Symplocarpus foetidus, to test the hypothesis that the extant species diversity of skunk cabbage is the result of a post-glacial range expansion from southern refugia during the Quaternary Ice Age. A total of 4041 base pairs (bp) of the chloroplast intergenic spacer region (cpDNA) was sequenced from 485 individuals sampled from glaciated (18 populations, 275 individuals) and unglaciated (14 populations, 210 individuals) regions east and west of the Appalachian Mountains. Haplotype number, haplotype diversity, and nucleotide diversity were calculated, and genetic variation within and among populations was assessed by analysis of molecular variance (AMOVA). The geographic pattern of genetic differentiation was further investigated with a spatial analysis of molecular variance (SAMOVA). A total of eight haplotypes and three genetic groups (SAMOVA) were recovered and a much higher haplotype number (eight haplotypes) and haplotype diversity (0.7425) was observed in unglaciated compared to glaciated populations (five haplotypes, haplotype diversity = 0.6099). All haplotypes found in glaciated regions represented a subset of haplotypes found in unglaciated regions. Haplotypes of S. foetidus likely diverged during the Tertiary (mid-Miocene and late Pliocene), predating the last glacial maximum (LGM). Predictions based on ecological niche modeling (ENM) suggested that there was considerably less suitable habitat for skunk cabbage during the LGM, and the habitat range was further south compared to the current distribution. Reduced variation and a subset of haplotypes in glaciated regions suggest a founder effect associated with range expansion via long-distance seed dispersal. Our results do not support the "Driftless Area" scenario for the northern refugium, rather the data suggest a "Northeastern" refugium near the southernmost extent of the LGM.

Complete chloroplast genome of Ulleung Island endemic flowering cherry, Prunus takesimensis (Rosaceae), in Korea.

Prunus takesimensis is an endemic flowering cherry on Ulleung Island, Korea. The complete chloroplast (cp) genome of P. takesimensis was generated by de novo assembly using whole-genome next-generation sequencing approach. The cp genome size was 157,948 bp in length consisting of four regions; large single copy region (85,959 bp), small single copy region (19,117 bp), and a pair of inverted repeat regions (26,436 bp). The genome contained a total of 128 genes, including 83 coding genes, 8 rRNA genes, and 37 tRNA genes. Phylogenetic analysis for 20 reported genomes within the family Rosaceae showed the monophyly of the genus Prunus including newly sequenced P. takesimensis.

Complete chloroplast genome of Prunus yedoensis Matsum.(Rosaceae), wild and endemic flowering cherry on Jeju Island, Korea.

The complete chloroplast genome sequences of the wild flowering cherry, Prunus yedoensis Matsum., which is native and endemic to Jeju Island, Korea, is reported in this study. The genome size is 157 786 bp in length with 36.7% GC content, which is composed of LSC region of 85 908 bp, SSC region of 19 120 bp and two IR copies of 26 379 bp each. The cp genome contains 131 genes, including 86 coding genes, 8 rRNA genes and 37 tRNA genes. The maximum likelihood analysis was conducted to verify a phylogenetic position of the newly sequenced cp genome of P. yedoensis using 11 representatives of complete cp genome sequences within the family Rosaceae. The genus Prunus exhibited monophyly and the result of the phylogenetic relationship agreed with the previous phylogenetic analyses within Rosaceae.

Molecular and morphological data reveal hybrid origin of wild Prunus yedoensis (Rosaceae) from Jeju Island, Korea: implications for the origin of the flowering cherry.

PREMISE OF THE STUDY: The subgenus Cerasus of the genus Prunus includes several popular ornamental flowering cherries. Of the hundreds of cultivars, P. ×yedoensis ('Somei-yoshino') is the most popular and familiar cultivar in Korea and Japan and is considered to be of hybrid origin. However, the hybrid origin of P. ×yedoensis and its relationship to wild P. yedoensis, naturally occurring on Jeju Island, Korea, are highly controversial. METHODS: We extensively sampled wild P. yedoensis, cultivated P. ×yedoensis, and numerous individuals from other species belonging to subgenus Cerasus on Jeju Island. Samples from 71 accessions, representing 13 species and one cultivar (P. ×yedoensis), were sequenced for nrDNA ITS/ETS (952 characters) and seven noncoding cpDNA regions (5421 characters) and subjected to maximum parsimony and maximum likelihood analysis. Additive polymorphisms in the ITS/ETS regions were confirmed by cloning amplicons from representative species. KEY RESULTS: The nuclear (ITS/ETS and G3pdh) and cpDNA data, along with several morphological characteristics, provide the first convincing evidence for the hybrid origin of wild P. yedoensis. The maternal parent was determined to be P. spachiana f. ascendens, while the paternal parent was unresolved from the taxonomically complex P. serrulata/P. sargentii clade. The presence of two kinds of ribotypes was confirmed by cloning, and the possible origin of cultivated P. ×yedoensis from wild populations on Jeju Island was also suggested. CONCLUSIONS: Bidirectional and multiple hybridization events were responsible for the origin of wild P. yedoensis. Extensive gene flow was documented in this study, suggesting an important role of reticulate evolution in subgenus Cerasus.

Flavihumibacter solisilvae sp. nov., isolated from forest soil.

A Gram-stain-positive, strictly aerobic, yellow colony-forming bacterium, designated strain 3-3(T), was isolated from forest soil of Bac Kan Province in Vietnam. Cells were non-motile rods without gliding motility, showing oxidase- and catalase-positive reactions. Growth was observed at 20-37 °C (optimum, 28 °C) and pH 5.5-9.5 (optimum, pH 7.5). The major cellular fatty acids were iso-C(15 : 0), iso-C(15 : 1) G and summed feature 3 (comprising C(16 : 1)ω6c and/or C(16 : 1)ω7c). Strain 3-3(T) contained phosphatidylethanoamine, three unidentified aminolipids and three unidentified lipids. The G+C content of the genomic DNA was 49.5 mol% and the only isoprenoid quinone detected was menaquinone 7 (MK-7). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain 3-3(T) formed a tight phylogenetic lineage with Flavihumibacter petaseus T41(T) with a bootstrap value of 100%. Strain 3-3(T) was related most closely to F. petaseus T41(T) with 97.3% 16S rRNA gene sequence similarity and the level of DNA-DNA relatedness between the two was 9.4±1.2%. Based on phenotypic, chemotaxonomic and molecular features, strain 3-3(T) represents a novel species of the genus Flavihumibacter, for which the name Flavihumibacter solisilvae sp. nov. is proposed. The type strain is 3-3(T) ( = KACC 17917(T) = JCM 19891(T)).