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Pelatantheria scolopendrifolia (Makino) Aver. is a critically endangered orchid species. In this study, an asymbiotic seed germination and development protocol to facilitate in situ and ex situ conservation of P. scolopendrifolia was determined. Seedlings were cultured in vitro on 1/2-strength Murashige & Skoog medium (1/2 MS), Phytamax Orchid Maintenance Medium (POM), Orchid BM1 Terrestrial Seed Germination & Stem Propagation Medium (OBTSG), or Orchid BM2 Terrestrial Stem propagation and Callus Medium (OBTSCM). Seeds started to germinate on all media except POM, with optimal growth on 1/2 MS medium, wherein 70% of seed germinated and produced roots and shoots within 12 weeks. Thus, we recommend culturing P. scolopendrifolia Seeds on 1/2 MS to facilitate germination and seed development, and in turn, in situ and ex situ conservation of this endangered species.

Prangos fedtschenkoi (Regel et Schmalh.) Korovin (Apiaceae) is an endemic species for moun-tainous Middle Asia, which is both a rare and useful plant. Organic extractions from this species are being used in pharmaceutics and cosmetology. In recent years, P. fedtschenkoi distribution area has considerably decreased, presumably, resulting from human activities such as agriculture, construction works, overgrazing and collection from wild for pharmaceutic purposes. Six populations were found in Uzbekistan and their genetic divergence and differentiation were studied with 10 inter-simple sequence repeat (ISSR) markers, selected out of 101. Totally 166 amplified ISSR fragments (loci) were revealed, of which 164 were polymorphic. Relatively mod-erate level of polymorphism was found at population level with polymorphic bands ranging from 27.71% to 47.59%. Mean P = 39.05%, Na=1.40,Ne=1.25, S.I. = 0.21, and He = 0.14 were revealed for all loci across six populations. AMOVA showed higher variation among populations (62%) than within them (38%). The Bayes-ian model determined 5 clusters, or genetic groups. The posteriori distribution of the Theta II estimator detected full model identifying high inbreeding, intensified by low gene flow (Nm = 0.3954). Mantel test confined pop-ulation 6 as distinct cluster corresponding to geographic remoteness (R = 0.5137, p ≤ 0.005). Results were used as the bases for developing conserve measures to restore populations.

The genus Cypripedium, known as lady's slipper orchid, has been considered to have a considerable marketability as a potted and garden plant with beautiful flowers; however, this species is becoming endangered and, in some places extinct, due to habitat destruction and illegal collections. As such, an optimized artificial propagation system is necessary for its conservation and horticultural cultivation. For the establishment of the in vitro proliferation of the endangered Cypripedium macranthos Sw., native to Korea, the effects of medium strength and sucrose concentrations on germination, protocorm formation and seedling growth were investigated through asymbiotic seed culture. The highest germination rate and protocorm formation rate were obtained with a 1/4 MS medium; higher MS medium strengths did not generate the favorable conditions required for germination and protocorm growth. The overall growth characteristics of roots and buds developed from protocorms were best in cultures of 1/4 MS medium. On this medium containing 10 g·L -1 sucrose, both the germination rate and protocorm formation rate increased significantly. The general growth properties of seedlings developed on the medium with 10 g·L -1 sucrose were best, showing the highest bud formation rate and root number. Our results demonstrate that the 1/4 MS medium, supplemented with 10 g·L -1 sucrose, could improve in vitro germination, and facilitate the growth of seedlings developed from the protocorms of C. macranthos Sw.

Reliable and accurate species identification is essential to establish a strategy for monitoring and manipulation of plant populations for conservation biology. However, identification of nonflowering plants is difficult. Based on recent advances in molecular biological techniques, various molecular identification methodologies have been proposed and applied in plant science, with a focus on medicinally valuable species. Lady’s slipper orchids (Cypripedium) are a representative terrestrial orchid group that includes many endangered species. Of 46 species, 3 (Cypripedium japonicum Thunb., Cypripedium macranthos Sw., and Cypripedium guttatum var. koreanum Nakai) were reported from Korea and categorised as endangered species or at risk of becoming endangered because of the rapidly decreasing population. We generated an accurate molecular identification system for these species using the sequence variation and species-specific SNPs of two plastid loci, rpoC2 and the IGS region between atpF and atpH. We selected the atpF-H region for molecular identification of the Korean Cypripedium and related Taiwan-endemic taxon Cypripedium formosanum Hayata. Cypripedium guttatum var. koreanum contained large deletions of over 300 bp, and could be distinguished by electrophoresis. For the other three species, we designed new primers based on a specific insertion (C. macranthos) and SNPs (C. japonicum and C. formosanum). We confirmed that molecular identification enables detection of each species using species-specific primers.

천연기념물 제 432호 제주 상효동의 한란 자생지(39 ha)를 대상으로 2013년 10월부터 2014년 2월까지 생물(식물상, 식생다양성 및 임분 구조) 및 비생물 환경(미기상, 수관열림도, 임내 광량 및 토양 온 습도)의 특성을 분석하였다. 한란 자생지의 일평균온도( $5.7^{\circ}C$ )는 서귀포시 보다 $3.3^{\circ}C$ 낮았고...

Saussurea pseudogracilis Kitam. is characterized by its slender leaves, small involucre and small number of involucral bracts compared to Saussurea gracilis Maxim. But some taxonomists insist that it is hard distinguih S. pseudogracilis from S. pseudogracilis. We tried to discern taxonomic identification of S. pseudogracilis by measuring and analyzing 45 qualitative characters and 42 quantitative characters on 165 specimen. S. pseudograilis was not distinguished from S. gracilis not only by qualitative characters but also by quantitative characters even on statistical analysis(PCA). Therefore, S. pseudogracilis is not independent taxonomic species and it is proper to consider S. pseudogracilis as a synonym of S. gracilis.

Many plant species in subalpine regions are under threat of extinction as a result of climate change. In this study, the genetic diversity and geographic differentiation of three regions and six populations of Primula farinosa subsp. modesta (Bisset & Moore) Pax in Korea were assessed using the ISSR (Inter Simple Sequence Repeat) marker. The average genetic diversity (P

Here we describe a new analytical method for the detection of two influenza A viruses by nitrocellulose membrane and magnetic sensors that employ a special frequency mixing technique. The combination of the nitrocellulose membrane and magnetic bead detection permits a rapid assay procedure and excludes two steps (the development of color and the stop reaction) required for usual immunochemical detection methods such as ELISA. Quantitative virus detection was performed using magnetic beads conjugated with secondary antibody. The results were compared with conventional assay methods and with a dot-blot assay with fluorescence compound (FITC). Under optimum conditions, our new assay procedure is capable of detecting picograms of virus per well. This new method combining the nitrocellulose membrane and magnetic bead detection reduces analytical time and allows stable and repeatable analyses of samples in point-of-care applications.

In this study, internal transcribed spacer (ITS) regions within the nuclear ribosomal DNA of KoreanIlex were analyzed in order to investigate any molecular evidence thatI. × wandoensis could serve as a putative hybrid betweenI. cornuta andI. integra. We also sought to clearly identify taxonomic relationships and problems caused by consecutive external morphological characters among taxa in the genus. We sequenced 20 clones fromI. × wandoensis and found these individuals displayed intra-genomic polymorphisms within ITS regions. The analysis of the clones clearly demonstrated the presence of discrete sequences from bothI. cornuta andI. integra, thereby confirmingI. × wandoensis is a species that was formed by crossingI. cornuta andI. integra. Lastly, the subgenusIlex, which contains the evergreen species, failed to form a monophyletic group in a strict consensus tree that was prepared based upon ITS regions.I. crenata var.microphylla in the subgenus KoreaIlex, which has presented taxonomic problems previously, formed an independent clade within the consensus tree, thereby showing distinction fromI. crenata.) Genetic discontinuity ofI. macropoda andI. Macropoda for.pseudo-macropoda individuals couldn't be confirmed.

Genetic variation of Ilex cornuta Lindley et Paxton was examined by sequence analyses of ITS for 65 individuals from Korea and China. The length of ITS 1 ranged from 253 to 259 bp. The 5.8S was 159 bp and ITS2 was observed to be 231 bp. A total of 8 different ITS types (Single Nucleotide Polymorphism haplotypes), which showed the difference of 1- 6 bp, were detected from 65 individuals. The sequence polymorphisms of ITS appeared at 9 different sites. All of four individuals collected at Daejeong-eup in Jeju-do exhibited different types, but all individuals from Naju-si and Muan-gun in Jeollanam-do were identical. The variation of ITS was higher in Jeju-do population than in inland population. Since 1. cornuta contains various types of ITS sequences, ITS analyses will provide important information on genetic diversity and conservation of this species.

The seed morphology and anatomy of 12 species, representing five sections of Thalictrum, were investigated by scanning electron microscopy (SEM) and light microscopy (LM) to evaluate seed features for use in systematic analysis. Considerable differences were observed in seed surface micromorphology and seed coat characteristics both among and within sections of the genus. MANOVA, MDA, and cluster analysis were used to unravel the morphology of seeds among the species. The characteristic seed features for species were found to be seed shape, size, testa thickness, exotestal cells and seed surface sculpture. The results indicated that the most prevalent seed coat sculpture among the studied species was reticulate and some species were reticulate at the margin and rugose in the center. Seed surface was without ornamentation and epidermal cells were mostly polygonal, rectangular or irregularly shaped. The testa thickness varied from two to six cell layers, and the exotesta was well characterized and rectangular to tangentially elongate in most of the species, except T. simplex and T. uchiyamae in which exotesta was poorly represented. It has been observed that species with a non-reticulate or poorly developed reticulate surface sculpture have a less protected seed coat and vice versa. The results of this study indicated that seed morphological features do not, in general, provide strong support to the sectional division of the genus, but that species within sections shared some comparable seed features.