• Korean Journal of Pharmacognosy
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• v.27 no.3
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• pp.224-230
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• 1996

To find pharmacologically active hybrids among the inter-order protoplast fusants of Lentinula edodes and Ginoderma lucidam the antitumor test was performed and the fusant P22 was selected among them. The hot water and alkaline extracts from the cultured mycelia of P22 were purified and separated into four fractions by DEAE-cellulose anion exchange chromatography. When a dose of 20 mg/kg/day of each fractions was injected into ICR mice by i.p., the tumor inhibition ratio of Fr. IV against solid sarcoma 180 was the higher than any other fraction. Fr. IV was a protein-bound polysaccharide which was composed of 69. 12% polysacchafide and 9.76% protein and the molecular weight of Fr. IV was $6.7{\times}10^4$ dalton.

• Journal of Plant Biotechnology
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• v.2 no.2
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• pp.55-60
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• 2000

Experiments initiated 30 years ago to obtain selectable markers have led to a series of studies of Trp biosynthesis and anthranilate synthase (AS) the control enzyme using largely plant tissue cultures since they have experimental properties that can be readily exploited. Enzymological and compound feeding studies provided evidence that AS is the control point in the Trp biosynthesis branch and that altering the AS feedback control by the selection of mutants resistant to the Trp analog 5-methyl-tryptophan (5MT) can lead to the overproduction of this important amino acid. Plants regenerated from these Trp overproducing lines of most species also had high free Trp levels but Nicotiana tabaum (tobacco) plants expressed the feedback altered AS only in cultured cells and not in the regenerated plants. further tests by transient and stable expression of the cloned promoter for the naturally occurring tobacco feedback-insensitive AS, denoted ASA2, confirmed the tissue culture specific nature of the expression control. The 5MT caused by the expression of a feedback-insensitive AS from tobacco has been used to select protoplast fusion hybrids with several species since the resistance is expressed dominantly. Recently the ASA2 gene has been used successfully as a selectable marker to select transformed Astragalus sinicus and Glycine max hairy roots induced by Agrobactetium rhizogenes. These results show that the ASA2y-subunit can interact with the y-subunit of another species to form active feedback-insensitive enzyme that may be useful for selecting transformed cells. Plastid DNA transformation of tobacco has also effectively expressed ASA2 in the compartment in which Trp biosynthesis is localized in the cell.

• KSBB Journal
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• v.5 no.2
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• pp.175-182
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• 1990

Protoplasts, isolated from leaf of N. tabacum NR-/SR+ and N. glottnosa were electrofused and divided with a plating efficiency of 30∼35% in AAPI 9M medium. Green callus lines were selected in protoplast-derived colonies on MSNO3 selection medium with 1.2mg/ml streptomycin sulfate on the basis of nitrate reductase proficiency and streptomycin resistance. Four putative hybrid plant lines regenerated from the green callus lines had intermediate morphology between that of parents with respect to floral shape, corolla length and ovate leaf blade. Zymograms of leaf peroxidase and esterase from these putative hybrid plant lines showed isozyme profiles derived from both parents and also, they exhibited additional and lost bands. Cytological analysis of two putative hybrid plant lines gave chromosome counts of 2n=66 in L22 and 2n=54 in L44 which were less than the expected number of N. tabacum(2n=48) and N. glutinosa(2n=24).

• Journal of Life Science
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• v.22 no.7
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• pp.871-879
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• 2012

Polyploids are a potentially important germplasm source in seedless citrus breeding program. Seedlessness is one of the most promising traits of commercial mandarin breeds that mandarin triploid hybrids possess permanently. The formation of new constant triploid hybrids can be recovered through diploid species hybridization from the fusion of divalent gametes at low frequencyor intra-and inter-ploidy crosses. However, extensive breeding work based on small $F_1$ hybrid seeds developed is impossible without a very effective aseptic methodology and ploidy event. In this study, in vitro embryo culture was employed to recover natural hybrids from monoembryonic diploid, open-pollinated mandarin. Flow cytometry was used to determine ploidy level. A total of 10,289 seeds were extracted from 792 fruits having approximately 13 seeds per fruit. Average frequency of small seeds developed was 7.1%, while the average frequency of small seeds per fruit were: 8.9% for 'Clementine' 10.2% for 'Harehime' 2.6% for 'Kamja' 3.1% for 'Pyunkyool' 2.8% for 'Sadookam' and 7.0% for 'Wilking' mandarin. Average size of a perfect seed was $49.52{\pm}0.07mm^2$ ('Clementine') while the small seed measured $7.95{\pm}0.04mm^2$ ('Clementine'), which was about 1/6 smaller than the perfect seed. In total, 731 small seeds were obtained and all of them contained only one embryo per seed. The efficiency of 'Clementine' was 14 times higher than 'Wilking' and more than 109 times higher than 'Pyunkyool'. The basic information on spontaneous polyploidy provides for the hybridization of constant triploids and increases the efficiency of conventional cross.

• 한국균학회소식:학술대회논문집
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• 2014.05a
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• pp.13-13
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• 2014

Two kinds of mushrooms, Gumji (金芝; Ganoderma) and Soji, were described in old book of Samguksagi (History of the three kingdoms; B.C 57~A.D 668; written by Bu Sik Kim in 1145) in Korea-dynasty. Many kinds of mushrooms were also described in more than 17 kinds of old books during Chosun-dynasty (1392~1910) in Korea. Nowadays, mushroom cultivation has been increased through out the world last decade years. Production of mushrooms has also been increased 10-20% and many varieties have been cultivated. Similar trends were also observed in Korea. Approximately two hundred commercial strains of 37 species in mushrooms were developed and distributed to cultivators. Somatic hybrid variety of oyster mushroom 'Wonhyeong-neutari' were developed by protoplast fusion, and distributed to grower in 1989. The fruiting body yield index of somatic hybrids of Pleurotus ranged between 27 and 155 compared to parental values of 100 and 138. In addition, more diverse mushroom varieties such as Phellinus baumi, Auricularia spp., Pleurotus ferulae, Hericium erinaceus, Hypsizigus marmoreus, Grifola frondosa, Agrocybe aegerita and Pleurotus cornucopiae have been attempted to cultivate in small scale cultivation. Production of mushrooms as food was 190,111 metric tons valued at 800 billion Korean Won (one trillion won if include mushroom factory products; 1dollar = 1,040 Won) in 2011. Major cultivated species are Pleurotus ostreatus, Pleurotus eryngii, Flammulina velutipes, Lentinula edodes, Agaricus bisporus, and Ganoderma lucidum, which cover 90% of total production. Since mushroom export was initiated from 1960 to 1980, the export and import of mushrooms have been increased in Korea. Technology developed for liquid spawn production and automatic cultivation systems lead to the reduction of the production cost resulting in the increasement of mushroom export. However some species were imported because of high production cost for these mushrooms requiring the effective cultivation methods. Developing of effective post-harvest system will be also directly related to mushroom export. In academic area, RDA scientists have been conducting mushroom genome projects. One of the main results is the whole genome sequencing of Flammulina velutipes for molecular breeding. An electrophoretic karyotype of of F. velutipes was obtained using CHEF with 7 chromosomes, with a total genome size of approximately 26.7 Mb. The mususcript of the genome of F. velutipes was published in PLOS ONE this year. For medicinal mushrooms, we have been conducting the genome research on Cordyceps and its related species for developing functional foods using this mushroom. In 2013, Korea Food and Drug Administraion (KFDA) approved Cordyceps mushroom for its value as an immune booster.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.69-76
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• 2011

This study describes the effect of myo-inositol on sustained cell division and plant regeneration from cotyledon-derived protoplast of cabbage (Brassica oleracea var. capitata). Freshly isolated protoplasts were cultured in modified Murashige and Skoog (MS) medium removed ammonia ions and containing $0.4\;mg\;l^{-1}$ thiamine HCl, $100\;mg\;l^{-1}$ myo-inositol, $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and several concentrations of myo-inositol (2, 4, 6, 8, 10% (w/v)) as an osmotic stabilizer. After 3 weeks of culture in the dark at $25^{\circ}C$, the plating efficiency of cabbage protoplasts reached to $22.5{\pm}2.9%$ when cultured in modified MS medium supplemented with $2\;mgl^{-1}$ 2,4-D, $0.5\;mgl^{-1}$ BA, $30\;gl^{-1}$ sucrose and 8% (w/v) of myo-inositol at a density of $2{\times}10^5$ protoplasts/ml. Rapidly growing cell colonies after 3 weeks of culture were transferred to the same culture medium removed osmoticum. To induce shoot regeneration from calluses, calluses with about 2 mm in diameter were transferred to the MS medium containing $2\;mgl^{-1}$ BA and $0.5\;mgl^{-1}$ NAA. After further three weeks of incubation onto the medium in the light, green shoots were formed on the surface of calluses at a frequency of 30%. Upon transfer to half-strength MS basal medium, roots were formed onto the bottom of regenerated shoots without auxin treatments. These regenerated plantlets were successfully acclimatized to soil transfer, grown to normal mature plants. The cabbage protoplast culture system established in this study could be applied for production of somatic hybrids or cybrids by asymmetric protoplast fusion and mass proliferation of elite somatic clones of cabbage.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.5 no.2
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• pp.141-149
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• 2004

The aim of this study was to test the validity of the hypothesis that the karyotypes of four species of Lepilemuridae were formed spontaneously from their ancestral hybrid karyotype. Hypothetical ancestral haploid Karyotype of Lepilemuridae is composed of 18 autosomes and X chromosomes. Lepilemur mustelinus karyotype has four tandem fused chromosomes and one Robertsonian translocated chromosome pairs. Lepilemur septentrionalis septentrionalis karyotype has only two pairs of translocated chromosomes. We reconstruct and suggest ancestral karyotype of LMU(ancLMU) and LSS(ancLSS), from which all four studied species were derived. Hybrids of ancLMU and ancLSS were formed and produce differently fused equilibrated gametes via circular form arrangement during gametogenesis. Five unit of trivalent homologous chromosome pairs were engaged in a circular form to give new gamete corresponding to the karyotype of L. dorsalis, orientation of one unit of trivalent was inversed in the circle to gave new gamete corresponding to the karyotype of L. leucopus. Seven homologous chromosome pairs were engaged in circular form to give haploid karyotype of Lepilemur ruficaudatus. Only one homologous chromosome pair is dissociated and the other chromosome pairs rearranged in the circle to form haploid karyotype of Lepilemur edwardsi. The new gametes could be produced from these circular forms. When the new gamete fertilized with the same type of gamete, The new homozygote is produced as existing L. dorsalis, L. leucopus, L. edwardsi and L. ruficaudatus. These results support the theory that new species could be formed in hybrid population through activated chromosome fusion, chromosome rearrangement in circular from at zygotene stage and production of equilibrated gametes to form homozygote new species.