• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.370-375
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• 2015

This experiment compared the regeneration conditions of the radiation mutant spray chrysanthemum 'purple ND'. The four different flower blooming stages (S1: 10% opened flower, S2: 30% opened flower, S3: 50% opened flower, and S4: 70% opened flower) and different petal parts (TBOP: the basal of petal and TEOP: the end of petal) were used to compare regeneration conditions between plants grown in MS medium supplemented with IAA and BAP. The highest adventitious shooting rate was identified in plants grown on the IAA $1.0mg{\cdot}L^{-1}$ and BAP $2.0mg{\cdot}L^{-1}$ when using the end of petal at the S2 stage. It displayed 79.2% regeneration and produced 33.4 shoots. Rooted plantlets were successfully established in the greenhouse, showing the same morphological characteristics of vegetative and reproductive organs with those of the mother plant. Flow cytometry analysis revealed no ploidy variation between the regenerated plants and the mother plant grown under greenhouse conditions.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.342-349
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• 2015

Kiwifruit is a new fruit crop that was commercialized in the late 1970s. Recently, its cultivation and consumption have increased rapidly worldwide. Kiwifruit is a dioecious, deciduous, and climbing plant having fruit with hairs and various flesh colors and a variation in ploidy level; however, the industry consists of very simple cultivars or genotypes. The need for efficient cultivar improvement together with the evolutional and biological perspectives based on unique plant characteristics, have recently encouraged genome analysis and bioinformatics application. The draft genome sequence and chloroplast genome sequence of kiwifruit were released in 2013 and 2015, respectively; and gene annotation has been in progress. Recently, transcriptome analysis has shifted from previous ESTs analysis to the RNA-seq platform for intensive exploration of controlled genetic expression and gene discovery involved in fruit ascorbic acid biosynthesis, flesh coloration, maturation, and vine bacterial canker tolerance. For improving conventional breeding efficiency, molecular marker development and genetic linkage map construction have advanced from basic approaches using RFLP, RAPD, and AFLP to the development of NGS-based SSR and SNP markers linked to agronomically important traits and the construction of highly saturated linkage maps. However, genome and transcriptome studies have been limited in Korea. In the near future, kiwifruit genome and transcriptome studies are expected to translate to the practical application of molecular breeding.

• Horticultural Science & Technology
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• v.29 no.6
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• pp.617-622
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• 2011

The study was conducted to determine the efficiency in producing spontaneous polyploids in some mandarin hybrids with different seed embryony. Seed formation by open pollination, frequency of spontaneous polyploids, and plant growth characteristics were evaluated in four mandarin hybrids with polyembryony such as 'Amakusa', 'Haruka', 'Hayaka', and 'Seminole' and two with monoembryony such as 'Benibae' and 'Harehime'. The mean number of the developed seeds per fruit was 10.0 and frequency of small seeds was 25.1%. Polyploids were selected from plants germinated in vitro by a flow cytometry and confirmed by chromosome analysis. One triploid was produced from 'Harehime', one tetraploid, 'Amakusa', and one tetrapoid, 'Benibae'. There were little differences in leaf shape, thickness, petiole length, and internode length between diploids and polyploids such as tri- or tetraploid. However, polyploids had larger stomata and lower density of stomata in abaxial epidermis than diploids. SPAS indicating chlorophyll content and photosynthetic rate were significantly affected by ploidy level. The results indicated that spontaneous polyploids might be produced by open pollination in some mandarin hybrids and monoembryony had higher frequency in polyploid occurrence than polyembryony.

• Development and Reproduction
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• v.21 no.1
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• pp.55-62
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• 2017

We used flowcytometry to ploidy verification after that investigate difference between diploid and spontaneous triploid through the truss dimension and classical dimension at crucian carp, Carassius auratus, crucian carp C. cuvieri and common carp, Cyprinus carpio collected from Hangang river, Hantangang river, Imjingang river, Geumgang river, Yeongsangang river and Nakdonggang river, Korea. There were significant differences among the three species for the truss dimensions anterior origin of dorsal fin $(2){\times}$ anterior origin of anal fin (5), $2{\times}$ anterior origin of pelvic fin (6), $2{\times}$ origin of pectoral fin (7), posterior origin of dorsal fin $(3){\times}5$, $3{\times}6$, and $3{\times}7$ (P<0.05). There were no significant differences among the three species in the truss dimensions dorsal fin length ($2{\times}9$) and eye diameter (ED)(P>0.05). On the other hand, there were no significant differences in the several classical dimensions of each species (P>0.05). Three classical dimensions, most anterior extension of the head $(1){\times}2$, $1{\times}6$ and $2{\times}$ most posterior scale in lateral line (4) did not differ between the C. auratus diploid and spontaneous triploid (P>0.05). Two classical dimensions, $1{\times}6$ and longest length between most anterior extension of the head and gill cover ($1{\times}8$) did not differ between the C. cuvieri diploid and spontaneous triploid (P>0.05). One classical dimensions, $1{\times}2$ did not differ between the common carp diploid and spontaneous triploid (P>0.05). There were significant differences in the each diploid and triploid species (P<0.05). These results suggest that the classification of each species and classification between diploid and spontaneous triploid morphometrical parameters used in this study are useful indices of morphometrical status in the each species from major river of Korea.

• Development and Reproduction
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• v.21 no.2
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• pp.193-204
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• 2017

The influence of triploidization on histological characteristics of retina, trunk kidney, liver and midgut tissue, and cell cycle of tail fin and gill tissue in far eastern catfish, Silurus asotus were analyzed. In the infertile triploid fish, the nucleus and/or cell size of secondary proximal tubule cells of trunk kidney, hepatocyte and midgut epithelium are much larger than those of the corresponding cells in the diploid fish (P<0.05). However, triploid tissue showed fewer number of outer nuclear layer in retina and nuclei in secondary proximal tubule of trunk kidney than those for diploid tissue. The mean percentages of the $G_l-$, the S- and the $G_2+M-phase$ fractions were 92.5%, 3.2% and 4.3% in tail fin tissue of diploid, and 93.4%, 2.6% and 4.0% in those of triploid, respectively. There were no significant differences in the percentages of each cell cycle fraction between diploid and triploid. The mean percentages of each phase fractions were 75.1%, 11.1% and 13.8% in gill tissue of diploid and 85.2%, 8.9% and 5.9% in those of triploid, respectively. The differences of cell cycle between tail fin tissue and gill tissue were statistically significant in diploid and triploid (P<0.05). Also, the differences between diploid and triploid were statistically significant in tail fin tissue and gill tissue (P<0.05). Cyclin D1 and cyclin E expressions were not significantly difference between gill tissue and tail fin tissue, and protein expressions of induced triploid were higher than those of diploid. Results from this study suggest that some characteristics in the triploid exhibiting larger cell and nucleus size with fewer number of cell than diploid can be used as an indicator in the identification of triploidization and ploidy level in far eastern catfish.

• Korean Journal of Plant Taxonomy
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• v.35 no.2
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• pp.129-142
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• 2005

Somatic chromosome numbers of 19 taxa of Korean Rubus was investigated. Subg. Anoplobatus (2 species), subg. Cylactis (1 species), subg. Idaeobatus (15 taxa) and subg. Malachobatus (1 species) are found in Korea. All taxa belonging to subg. Idaeobatus except for R. parvifolius which shows tetrapolid and hexaploid are diploid. The basic chromosome number of the genus was x=7. New chromosome numbers for 5 taxa were reported here: R. hongnoensis of Jeju-island endemic species, 2n=14; R. longisepalus, 2n=14; R. longisepalus var. tozawai, 2n=14; R. parvifolius, 2n=28; R. parvifolius var. taquetii, 2n=28. The rest 12 taxa except for R. coreanus Miq was well counted as 2n=14 and well consistent with previous reports from China and Japan. Our new chromosome level for R. parvifolius as 6x may indicate that speciation by polyploidization has occurred within Korean population. Unlikely to Japanese population (2n=42), Korean population of R. buergeri has same ploidy level with Taiwanese population as 2n=56.

• Horticultural Science & Technology
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• v.34 no.6
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• pp.966-976
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• 2016

A new bright yellow gerbera (Gerbera hybrida Hort.) cultivar, 'Joyful', was released by the Flower Research Institute of Gyeongsangnam-do Agricultural Research & Extension Service in 2013. 'Joyful' was developed in Changwon from 2005 to 2006 and is a cross between 'Chowoo', a semi-double type gerbera cultivar with yellow-orange colored flowers, and 'Focus', a semi-double type cultivar with yellow colored flowers. The characteristics trials for selection were conducted from 2007 to 2009. 'Joyful' has large, semi-double type flowers with yellow (RHS, 12-A) ray florets and a brown central part. It also has a stable flower shape, strong peduncles, and a vase life of 12.8 days. The average flower yield of 'Joyful' was determined in greenhouse trials carried out from 2008 to 2010 and is 49.2 stems per plant per year. 'Joyful' was demonstrated to be a new cultivar based on ploidy tests and RAPD analysis and was registered (Grant No. 4574) to the Korea Seed and Variety Service (KSVS) for commercialization in 2013.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.656-664
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• 2015

To produce high quality watermelon, three tetraploid watermelon breeding lines (‘SA03-1’, ‘SA06-1’ and ‘SB01-1’) were developed by treatment with different chromosome doubling reagents. To identify the optimal tetraploid inductive conditions, the three watermelon breeding lines were selected by counting the number of doubled chloroplasts in guard cells. Tetraploid induction rates differed depending on the genotypes and treatment with doubling reagents. However, the highest induction rate occurred with 1.0% colchicine (82.2%). These putative tetraploid lines were re-confirmed for ploidy using flow cytometric analysis and chromosome counting. The internode length of the tetraploid breeding lines was different when the leaf size was larger in all three tetraploid lines compared to their diploids. The fruit weight of the tetraploid fruits for ‘SA03-1’ and ‘SB01-1’ was lower than for their diploid, and the rind thickness and total sugar content (°Brix) of tetraploid SB01-1 were significantly different from those of its diploid. Tetraploid lines were sterile, yielded a lower number of seeds per fruit for ‘SA03-1’ (21), ‘SA06-1’ (62), and ‘SB01-1’ (34.7), and the seeds were larger and thicker than those of their diploids. These tetraploid breeding results will be useful for breeding new seedless watermelon cultivars.

• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1347-1354
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• 2006

The aims of this study were to establish a simple and effective method for isolating male germline stem cells (GSCs), and to test the possibility of using these cells as a new approach for male infertility treatment. Testes obtained from neonatal and adult mice were manually decapsulated. GSCs were collected from seminiferous tubules by a two-step enzyme digestion method and plated on gelatin-coated dishes. Over 5-7 days of culture, GSCs obtained from neonates and adults gave rise to large multicellular colonies that were subsequently grown for 10 passages. During in vitro proliferation, oct-4 and two immunological markers (Integrin ${\beta}1,\;{\alpha}6$) for GSCs were highly expressed in the cell colonies. During another culture period of 6 weeks to differentiate to later stage germ cells, the expression of oct-4 mRNA decreased in GSCs and Sertoli cells encapsulated with calcium alginate, but the expression of c-kit and testis-specific histone protein 2B(TH2B) mRNA as well as the localization of c-kit protein was increased. Expression of transition protein (TP-l) and localization of peanut agglutinin were not seen until 3 weeks after culturing, and appeared by 6 weeks of culture. The putative spermatids derived from GSCs supported embryonic development up to the blastocyst stage with normal chromosomal ploidy after chemical activation. Thus, GSCs isolated from neonatal and adult mouse testes were able to be maintained and proliferated in our simple culture conditions. These GSCs have the potential to differentiate into haploid germ cells during another long-term culture.

• Korean Journal of Agricultural Science
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• v.39 no.4
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• pp.497-502
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• 2012

Since the year 2000, the production and consumption of broccoli have rapidly increased in Korea. And, the average production area and amount were about 1,700 ha and 29,000 ton for the past 5 years. Even with the increase of these cultivation and consumption, more than 95% of the broccoli seeds are currently imported from foreign countries such as Japan and Netherlands. Therefore, development of domestic broccoli varieties is needed to relieve Korean farmers' production cost for broccoli. In this situation, National Institute of Horticultural and Herbal Science (NIHHS) of Korea has tried to develop F1 hybrid varieties from elite lines that were obtained by microspore cultivation method from 2008. As the results, about 850 lines of broccoli were obtained and self-pollinated. Then their ploidy levels of the genome were confirmed to select double haploid (D.H.) lines. And the D.H. lines' horticultural traits were evaluated in open field. After the selection of 17 elite D.H. lines, they were cross-pollinated with a male sterile (MS) line to produce F1 hybrid seeds. After 2 to 3 years field trials of these F1 hybrid varieties at the area of Suwon, Gangneung, and Jeju respectively, two hybrid varieties such as 'Wongyo8011' and 'Wongyo8012' are selected for the application of variety protection. With these 4 years of research, we found that the microspore cultivation method is a powerful tool for the conventional breeding program, especially for the development of various inbred lines and even F1 hybrid varieties in short time.