Taxonomic relevance of petiole anatomical and micro-morphological characteristics of Clematis L. (Ranunculaceae) taxa from South Korea.

We assessed the micro-morphological and anatomical structures of the petioles of 19 Clematis taxa from South Korea. The petiole surface features were observed with the help of stereomicroscopy and scanning electron microscopy, and the anatomical features are studied via microtomy and light microscopy. The results of this study showed that the presence/absence and abundance of trichomes, petiole cross-section outlines, upper surface wings and grooves, and the number of vascular bundles were useful for species discrimination in Clematis. Among the studied taxa, C. hexapetala was the only species with a glabrous petiole surface. Two types of trichomes were observed in the other 18 taxa: long, non-glandular and flagelliform trichomes and short, glandular capitate trichomes. We found four to six major vascular bundles and a maximum of eight interfascicular vascular bundles (C. heracleifolia and C. urticifolia) in the 19 taxa. A cluster analysis based on UPGMA identified six clusters with 18 nodes. Although the number of taxa investigated was limited, taxa from the sections Tubulosae, Viorna, and Astragene clustered with each other in the UPGMA phenogram due to the overall similarity of petiole features. Based on this observation, we can conclude that most of the petiole features are limited to the species level, and thus, the data obtained could be used as descriptive and/or diagnostic features for particular taxa, which may be useful for the investigation of problematic taxa in the genus.

Complete chloroplast genome of Clematis taeguensis (Ranunculaceae), an endemic species from South Korea.

The complete chloroplast (cp) genome sequence of Clematis taeguensis Y.N.Lee (Ranunculaceae) was determined to be 159,534 bp in length, consisting of large (79,326 bp) and small (18,338 bp) single-copy regions and a pair of identical inverted repeats (30,935 bp). The genome contains 92 protein-coding genes, 32 tRNA genes, 8 rRNA genes, and 1 pseudogene (infA). Phylogenetic analysis of 19 taxa inferred from the chloroplast genome showed a relationship with C. taeguensis, which is also recognized as a species endemic to the Korean Peninsula. The complete cp genome sequence of C. taeguensis reported here provides important information for future phylogenetic and evolutionary studies in Ranunculaceae.

Achene Morphology and Anatomy of Clematis L. (Ranunculaceae) in Korea and Its Taxonomic Implications.

Clematis, a widely distributed genus in Ranunculaceae, is one of the most difficult groups of taxa in the family from a taxonomic point of view. A comprehensive study on achene morphology and the anatomy of 19 taxa of Clematis from Korea was carried out using scanning electron and light microscopy to evaluate the taxonomic significance of achene characters. Clematis achenes are elliptical, obovate or fusiform in shape, light yellow or brown to black in color and completely or sparsely covered with hairs. The permanent style is elongated and plumose in all the studied taxa except C. brachyura. We found that the size, indument, permanent style, surface sculpture, shape in cross-section, and nature and thickness of the exocarp, and endocarp were valuable achene features for species delimitation and may contribute to the unraveling of the taxonomic problems in the genus Clematis. One-way analysis of variance (ANOVA) indicated that the quantitative achene variables among the species were highly significant (p < 0.001). Principal component analyses based on seven quantitative characters and UPGMA (unweighted pair-group method with arithmetic mean) analysis based on seven quantitative and 18 qualitative characters also signify the utility of achene features for taxonomic discriminations of the Clematis taxa within the genus. Similar to other morphological characters in the genus Clematis, achene morphological and anatomical characters with the limited taxonomic value alone cannot be expected to resolve the infrageneric relationships but certain achene features combined with other morphological features could be useful as an alternative means of determining the infrageneric relationships within the genus.

Gametophyte and embryonic ontogeny: understanding the reproductive calendar of Cypripedium japonicum Thunb. (Cypripedoideae, Orchidaceae), a lady's slipper orchid endemic to East Asia.

BACKGROUND: The genus Cypripedium L. is one of the five genera of the subfamily Cypripedioideae, members of which are commonly known as lady's slipper orchids. Cypripedium japonicum is a perennial herb native to East Asia, specifically China, Japan, and Korea. Due to its limited distribution, the species is included in the Endangered category of the IUCN Red List. RESULTS: We investigated gametophyte development, including complete embryogenesis, in C. japonicum. The complete reproductive cycle is presented based on our observations. Anther development begins under the soil, and meiosis of pollen mother cells begins 3 weeks before anthesis, possibly during early April. The megaspore mother cells develop just after pollination in early May and mature in mid-late June. The pattern of embryo sac formation is bisporic, and there are six nuclei: three forming the egg apparatus, two polar nuclei, and an antipodal cell in the mature embryo sac. Triple fertilization results in the endosperm nucleus, which degenerates when the proembryo reaches the eight-to-sixteen-cell stage. CONCLUSION: Our overall comparisons of the features of gametophyte and embryo development in C. japonicum suggest that previous reports on the embryology of Cypripedium are not sufficient for characterization of the entire genus. Based on the available information, a reproductive calendar showing the key reproductive events leading to embryo formation has been prepared.

Comparative wood anatomy of Korean Viburnum L. (Adoxaceae) and its taxonomic implication.

Comparative wood anatomy of Viburnum was carried out to understand the differences in wood features amongst the species which might be useful for taxonomic discrimination in the genus. Altogether, nine taxa belonging to five clades were investigated using a sliding microtome and light microscopy. The growth rings are well represented and earlywood and latewood are distinguishable in cross-section. Some of the important wood features include angular, oval and rounded vessels with scalariform perforation plates, opposite to scalariform inter-vessel pitting, rounded pits with slit-like apertures, thick-walled xylem tracheids with simple, rounded bordered pits, diffuse axial parenchyma, uni- and multiseriate rays, 2-4 cells wide. In general, there is a remarkable uniformity in the qualitative wood features in Viburnum species, although quantitative measurement showed some disparities. The most significant quantitative wood variables which might be useful for taxonomic groupings of the species comprise a frequency of vessels and rays, the diameter of the vessels and tracheids in the radial and tangential planes and height and width of rays in the tangential plane.

Correction to: The embryological insight into the relationship between Forsythia and Abeliophyllum (Forsythieae, Oleaceae).

In the original publication of the article, species name.

The embryological insight into the relationship between Forsythia and Abeliophyllum (Forsythieae, Oleaceae).

Forsythia saxatilis, a Korean native Forsythia, is recognized as an endangered species in the Korean Red List. We carried out a comprehensive embryological investigation, including a study of the pericarp development, of F. saxatilis and compared it with previously acquired information on the family Oleaceae and also with Abeliophyllum distichum, a close relative of Forsythia. Our results revealed that several embryological features of Forsythia are exclusively comparable with the family Oleaceae, particularly in relation to A. distichum. Despite the differences in fruit types and pericarp ontogeny, Forsythia shares some unique embryological features with Abeliophyllum, for instance the basic type of anther wall development, formation of the nucellar cap, a long micropyle, formation of the obturator and hypostase, and the crushed endotesta. Furthermore, the loculicidal capsule of Forsythia seems to be primitive within the tribe Forsythieae and samara of Abeliophyllum might be evolved from it. In conclusion, a considerable number of synapomorphies of embryological characters supports recent molecular reports and provide additional embryological evidence for the sister-group relationship of Forsythia and Abeliophyllum.

Allelopathic and Autotoxic Effects of Medicago sativa-Derived Allelochemicals.

In this study, the allelopathic properties of Medicago sativa on different weeds were investigated under in vitro conditions. The compounds involved in the autotoxicity of M. sativa were analyzed using high-performance liquid chromatography. The extracts of all concentrations inhibited the growth of the calluses of Digitaria ciliaris, Chenopodium album, Amaranthus lividus, Portulaca oleracea, and Commelina communis. Six allelopathic compounds in alfalfa were identified and quantified, and the most predominant phenolic compounds were salicylic acid and p-hydroxybenzoic acid. Various concentrations (10-2, 10-3, and 10-5 M) of all the tested phenolic compounds exerted inhibitory effects on callus fresh weight. Rutin, salicylic acid, scopoletin, and quercetin significantly inhibited alfalfa seed germination. Of the seven identified saponins, medicagenic acid saponins exhibited the highest autotoxic effect and significantly lowered seed germination rate. Principal component analysis showed that the phenolic compounds and saponin composition significantly contributed to the different variables. The highly phytotoxic properties of the alfalfa-derived phenolic compounds and saponins indicate that these phytochemicals can be a potential source of bioherbicides.

Leaf anatomy and its implications for phylogenetic relationships in Taxaceae s. l.

The comparative study on leaf anatomy and stomata structures of six genera of Taxaceae s. l. was conducted. Leaf anatomical structures were very comparable to each other in tissue shape and their arrangements. Taxus, Austrotaxus, and Pseudotaxus have no foliar resin canal, whereas Amentotaxus, Cephalotaxus, and Torreya have a single resin canal located below the vascular bundle. Among them, Torreya was unique with thick-walled, almost round sclerenchymatous epidermal cells. In addition, Amentotaxus and Torreya were comprised of some fiber cells around the vascular bundle. Also, Amentotaxus resembled Cephalotaxus harringtonia and its var. nana because they have discontinuous fibrous hypodermis. However, C. fortunei lacked the same kind of cells. Stomata were arranged in two stomatal bands separated by a mid-vein. The most unique stomatal structure was of Taxus with papillose accessory cells forming stomatal apparatus and of Torreya with deeply seated stomata covered with a special filament structure. Some morphological and molecular studies have already been discussed for the alternative classification of taxad genera into different minor families. The present study is also similar to these hypotheses because each genus has their own individuality in anatomical structure and stomata morphology. In conclusion, these differences in leaf and stomata morphology neither strongly support the two tribes in Taxaceae nor fairly recognize the monogeneric family, Cephalotaxaceae. Rather, it might support an alternative classification of taxad genera in different minor families or a single family Taxaceae including Cephalotaxus. In this study, we would prefer the latter one because there is no clear reason to separate Cephalotaxus from the rest genera of Taxaceae. Therefore, Taxaceae should be redefined with broad circumscriptions including Cephalotaxus.