• Journal of Korean Society of Forest Science
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• v.31 no.1
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• pp.48-52
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• 1976

In order to investigate the effects of seed weight, size (length and thickness), and maturation period (early maturing seed or late maturing seed) on growth of first-year seedlings in Pinus koraiensis, pot cultivation experiment with seeds collected from Gangweon University Forest was carried out under relatively controlled environment condition. At the end of one growing season, not only fresh weight, length, diameter (at underside of cotyledon) and root length of seedlings, but also number and length of cotyledon were measured. Results are as follows; 1. Germination percentage is independent of weight and size of seeds, but it was influenced by maturation period, that is, late maturing seeds surpass early maturing one in germination percentage. 2. Germination percentage is affected by maturation period of seeds in 18.3% of contribution rate. 3. Each growth of first-year seedlings is influenced considerably by the differences of weight, length and thickness of seeds, that is, heavy and large seeds produce larger seedlings than do light and small seeds. Particularly, fresh weight, diameter, length of cotyledon are significantly affected by differences of seed weight and size (length and thickness) in 50-90% of contribution rate. 4. Differences of seed weight and thickness have a tendency to affect more on growth of first-year seedlings than do differences of seed length. 5. Seedlings from late maturing seeds are superior than those from early maturing one in fresh weight, diameter of seedlings and length of cotyledon, but it's influence is a little.

• Horticultural Science & Technology
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• v.19 no.3
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• pp.315-319
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• 2001

The study was carried out to develop a simple and efficient system to regenerate plants from cotyledon and hypocotyl tissues of Chinese cabbage (Brassica campestris L. ssp. pekinensis cv Seoul). Among the various combinations of naphthalene acetic acid (NAA) and 6-benzyladenine (BA) tested, the best shoot induction medium for cotyledon, with 2.67 shoots per explants, contained $2.0mg{\cdot}L^{-1}$ NAA, $1.0mg{\cdot}L^{-1}$ BA and $16.7mg{\cdot}L^{-1}$ $AgNO_3$. The shoot induction medium with $1.0mg{\cdot}L^{-1}$ NAA, $5.0mg{\cdot}L^{-1}$ BA and $16.7mg{\cdot}L^{-1}$ $AgNO_3$, was best for shoot induction from hypocotyl explants, with 1.87 shoots per explants. After shoot induction, regenerated shoots were excised and rooted on rooting medium. Rooted plantlets were then hardened in the high humidity growth chamber and transplanted to pots, and then grown in the greenhouse. Regenerated plants appeared phenotypically normal and there were no changes in chromosome number.

• Proceedings of the Ginseng society Conference
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• 1974.09a
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• pp.9-22
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• 1974

Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

• Journal of Plant Biology
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• v.39 no.1
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• pp.71-77
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• 1996

Embryogenic callus was selected from callus induced from hypocotyl segment cultures of Daucus carota seedlings on MS basal medium supplemented with 1.0 mg/L 2,4-D. Cell clumps prepared from the embryogenic callus were transferred to MS medium without 2,4-D for somatic embryo development. Cotyledonary abnormalities were frequently observed on somatic embryos developed after the incubation of cell clumps in MS basal medium with 1.0 mg/L 2,4-D for one week and then subculture in the same medium but without 2.4-D for two weeks. The percentage of abnormalities was as follows: 5% one cotyledon, 21% three cotyledons, 6% four cotyledons, 5% five cotyledons, 0.2% six cotyledons and 1% trumpet-like cotyledons. On the other hand, the normal somatic embryo with two cotyledons appeared at 63%. The germination rate of somatic embryos was higher in two cotyledon somatic embryos than in multicotyledonary embryos. Trumpet-like somatic embryos did not germinate normally showing limited elongation and enlargement of roots and cotyledons without shoot development. From anatomical examination circular procambium in the root of somatic embryo began to branch around the middle regions of the hypocotyl which extended into the cotyledons through the cotyledonary nodes and the number of branched procambial strands in hypocotyl was equal to the number of cotyledons. Monocotyledonous somatic embryo always had larger cotyledon than that of somatic embryos with multicotyledons and had horseshoe-like cotyledons where the procambium was of the same structure.ucture.

• Journal of Plant Biotechnology
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• v.5 no.4
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• pp.245-250
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• 2003

An efficient procedure was developed for adventitious shoot bud induction and plantlet regeneration from various explants of the ten genotypes of Pepper (Capsicum annuum L.) using Thidiazuron (TDZ). Among various treatments at 1.0-3.0 mg/L TDZ Induced maximum number of adventitious shoots depending upon the explant type and genotype compared to other treatments. Among the explants tested, leaf induced maximum number of adventitious shoots than the cotyledons. TDZ-mediated organo-genesis was possible in 10 pepper cultivars, the extent of the response being genotype-dependent. Of the ten genotypes tested, C. annuum cvs CA960, $G_4$ and X-235 were produced maximum number of adventitious shoots and Sell was the least, and all other genotypes gave moderate response. Elongation of multiple shoots was observed on medium supplemented with SA (0.05 mg/L) in combination of IAA (0.05 mg/L). Differences in ability for in vitro shoot regeneration and elongation depend upon the variety and explant type. The elongated shoots were success. Fully rooted on MS medium containing at 1.0 mG/L IAA. Plantlets regenerated from different explants of ten genotypes were found to be diploid (2n=24) and were devoid of any chromosomal aberrations. Regenerated plants were successfully established in soil where 85-90% of them developed into morphologically normal and fertile plants.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.249-255
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• 2006

Effects of ethylene inhibitors like silver nitrate $(AgNO_3)$, cobalt chloride $(CoCl_2)$ and Salicylic acid (SA) on multiple shoot induction and their impact on ethylene production using embryonal cotyledon cultures of Cucumis sativus L. were examined. The optimum concentration of $AgNO_3\;(40{\mu}M),\;CoCl_2\;(20{\mu}M)\;and\;SA\;(20{\mu}M)$, separately, induced maximum number of shoots on Murashige and Skoog's (MS) medium supplemented optimally with $4.44{\mu}M$ BA and $0.25{\mu}M$ NAA. Among the three ethylene inhibitors tested, $AgNO_3$ produced maximum number of shoots when compared to $CoCl_2$ and SA Ethylene production was monitored in all the treatments with $AgNO_3/CoCl_2/SA$ and it was observed that the treatment with $AgNO_3$ alone showed increase in ethylene production when compared to $CoCl_2$ and SA Even though ethylene concentration was the highest in $AgNO_3$ treated explants, maximum number of shoots was obtained.

• Journal of Plant Biotechnology
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• v.6 no.2
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• pp.91-96
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• 2004

The use of liquid-medium-based procedure relative to the solid media led to a 4.5-fold increase in the number of cotyledon-stage embryos. The most efficient system for multiplication and regeneration of somatic embryos was CP6 procedure with the media MSD40/MSD20/MSM6AC/FNL0S3S3GM. However, the rate of regeneration was lower. About 71％ of the embryos with dicotyledon were continued to develop the roots after desiccation treatment and 92％ of the germinated embryos produced shoots in 10 days. Of the four morphologically different types of embryos, dicotyledonous ones showed a high frequency of conversion, while only a few with fused and horn type cotyledon developed shoots. Mature somatic embryos were desiccated in empty petri dishes for 12-72 h. Embryo survival rate was the highest after 12 h of desiccation, but maximal germination was observed at 24 h. After desiccation, they were placed on MS medium without growth regulators for germination. Germinating embryos were transferred to small pots with vermiculite for plant regeneration. The etiolating the plants during the growth was resolved to add 1％ activated charcoal on hormone-free MS medium.

• Journal of Life Science
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• v.11 no.2
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• pp.166-172
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• 2001

Various types of the seeding of bottle gourd widely used as a rootstock of watermelon has been required to satisfy the farmers need. The study was done to determine the effect of light quality of blue, red and far-red lights treated with light emitting diodes on growth and morphology of bottle goured seeding and chlorophyll content of its cotyledons. The lights were treated in the growth chamber for 7 days to the seeding elapsed 8 days after sowing under natural condition, and 64 hole trays with commercial bedsoil. Plant height, length and diameter of hypocotyl, leaf area of cotyledon and first true leaf, its leaf length, number of true leaves, fraction and total dry weight were measured. Red light shortened and slenderized the hypocotyl, which lengthened by far-red light and thickened by blue light. Plant height was declined in order of far-red light treatment, blue and red lights mainly due to difference of hypocotyl length, Area and length of the first true leaf became smaller and shorter under far-red light than under the other lights. However, blue light increased leaf area of cotyledons. Two cultivars cv. Yongjadaemok and cv. Kunghap had different response to the light treatments in total seedling dry weight(dw); far-red and red light treatments showed the greatest and the least dw of hypocotyl, respectively, while blue and red lights did the greatest dw of the other organs. Among the ratio of each organ dw to total dw, those of hypocotyl and true leaves were different between the light treatments; the highest ratio of hypocotyl dw to total dw was observed in far-red light treatment but the lowest was in red light treatment. Those of the true leaves were the lowest in far-red light and similar response in blue or red light treatment. Chlorophyll content of cotyledons was decreased in order of red light treatment, blue and far-red lights, meaning that short period light treatment may influence photosynthesis of seeding and afterward its growth.

• Journal of Plant Biotechnology
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• v.3 no.2
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• pp.95-100
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• 2001

To select the section with shoot formation ability, the calli and shoot formation from three sections (first leaf including cotyledonary node, hypocotyl and cotyledon explants) of 5-days-seedlings of soybean were induced on MS medium supplemented with 1.0 mg/L BAP, 3% sucrose, and 0.3% gelrite for one month. The first leaf section exhibited the highest shoot formation rate (51%), followed the hypocotyl section (10%) and the cotyledon section (0%). The shoot formation rates and shoot number of the four excised sections (whole first leaf, a half of the first leaf, a third of the first leaf and only node) of the first leaf were also investigated on the same medium. A half of the first leaf explant and the third of the first leaf explant had higher shoot formation rates (76-80%) and numbers (3-4 / explants) than those in other two explants. Effects of six cytokinins (kinetin, zeatin, BAP, 2iP, PBA, and TDZ) on shoot formation were determined, using the half of the first leaf explants. Zeatin (1.0 mg/L) exhibited the highest in shoot formation rate (94%) and numbers (8 / explant). In addition, the combined effects of three cytokinins (zeatin, BAP, and TDZ; 0.5, 1.0, 2.0 mg/L, respectively) and an auxin (IAA; 0.0, 0.5, 1.0, 2.0 mg/L) were determined. The combination (1:1, v/v) of zeatin (1.0 mg/L) and IAA (1.0 mg/L) exhibited the highest in shoot formation rate (96%) and numbers (16 / explant), twice more than zeatin (1.0 mg/L) alone. The shoot cuttings were transferred and cultivated on the rooting media supplemented with only auxin, IBA at various concentrations. The highest root formation (8 / shoot) was achieved on the medium supplemented with 1.5 mg/L. After 4 weeks of cultivation, the plantlets with an extensive root system were transplanted in pots with a soil mixture of vermiculite and fine sand. Transferred to field, about 75% of the plantlets survived.

• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.243-248
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• 2006

A optimal procedure for plant production via repetitive secondary somatic embryogenesis in Tilia amurensis is described. Somatic embryos were induced directly from the culture of zygotic embryos on medium with 1.0 mg/l 2,4.-D. Repetitive secondary somatic embryos formed on the surface of the cotyledons and hypocotyls except for the radicles when explants of somatic embryos were cultured on medium with 4.0 mg/l 2,4-D. The highest frequency of secondary embryo-genesis was obtained in the cotyledons (90%) and hypocotyls(83.33%) on MS medium with 1.0 mg/L 2,4-D. The average number of secondary embryos per explant was 25.74 in cotyledon and 24.92 in hypocotyl. When the cotyledon and hypocotyl segments from somatic embryos at different developmental stages were cultured on MS medium with 1.0 mg/L 2,4-D, the highest frequency of secondary embryogenesis was obtained from late cotyledonary secondary embryos. Somatic embryos were transferred to MS basal medium and then they germinated within 2 to 4 weeks of culture. Germinated somatic embryos grew normally into plantlets on WPM medium, producing new shoots. The converted plantlets were acclimatized on artificial soil mixture. These results indicate that the repetitive secondary somatic embryogenesis in T amurensis can offer the possibility to use in vitro culture system for the micropropagation.