• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.26 no.1
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• pp.69-89
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• 1981

Stevia (Stevia rebaudiana Bertoni) is a perennial herb widely distributed in the mountainous area of Paraguay. It belongs to the family Compositae and contains 6 to 12 percent stevioside in the leaves. Stevioside is a glucoside having similar sweetening character to surgar and the degree of sweetness is approximately 300 times of sugar. Since Korea does not produce any sugar crops, and the synthetic sweetenings are potentially hazardous for health, it is rather urgent to develop an economical new sweetener. Consequently, the current experiments are conducted to establish cultural practices of stevia, a new sweetening herbs, introduced into Korea in 1973 and the results are summarized as followings: 1. Days from transplanting of cuttings to the flower bud formation of 6 stevia lines were similar among daylengths of 8, 10 and 12 hours, but it was much greater at daylengths of 14 or 24 hour and varietal differences were noticable. All lines were photosensitive, but a line, 77013, was the most sensitive and 77067 and Suweon 2 were less sensitive to daylength. 2. Critical daylength of all lines seemed to be approximately 12 hours. Growth of plants was severely retarded at daylengths less than 12 hours. 3. Cutting were responded to short daylength before rooting. Number of days from transplanting to flower bud formation of 40-day old cuttings in the nursery bed was 20 days and it was delayed as duration of nursery were shorter. 4. Number of days from emergence to flower bud formation was shortest at short day treatment from 20 days after emergence. It was became longer as initiation of short day treatment was earlier or later than 20 days. 5. Plant height, number of branches, and top dry weight of stevia were reduced as cutting date was delayed from March 20 to May 20. The highest yield of dry leaf was obtained at nursery duration of 40-50 days in march 20 cutting, 30-40 days in April 20 cutting, and 30 days in May 20 cutting. 6. An asymptotic relationship was observed between plant population and leaf dry weight. Yield of dry leaf increased rapidly as plant population increased from 5,000 to 10,000 plants/10a with a reduced increasing rate from 10,000 to 20,000 plants/l0a, and levelled off at the plant population higher than 20,000 plants/l0a. 7. Stevia was adaptable in Suweon, Chengju, Mokpo and Jeju and drought was one of the main factors reducing yield of dry leaf. Yield of dry leaf was reduced significantly (approximately 30%) at June 20 transplanting compared to optimum transplanting. 8. Yield of dry leaf was higher in a vinyl house compared to unprotected control at long daylength or natural daylength except at short day treatment at March 20. Higher temperature ill a vinyl house does not have benefital effects at April 20 transplanting. 9. The highest content of stevioside was noted at the upper leaves of the plant but the lowest was measured at the plant parts of 20cm above ground. Leaf dry weight and stevioside yield was mainly contributed by the plant parts of 60 to 120cm above ground but the varietal differences were also significant. 10. Delayed harvest by the time of flower bud formation increased leaf dry weight remarkably. However, there were insignificant changes of yield as harvests were made at any time after flower bud formation. Content of stevioside was highest at the time of flower bud formation and earlier or later harvest than this time was low in its content. The optimum harvesting time determined by leaf dry weight and stevioside content was the periods from flower bud formation to right before flowering that would be the period from September 10 to September 15 in Suweon area. 11. Stevioside and rebaudioside content in the leaves of Stevia varieties were ranged from 5.4% to 14.3% and 1.5% to 8.3% respectively. However, no definit relationships between stevioside and rebaudioside were observed in these particular experiments.

• Journal of Korean Society of Forest Science
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• v.42 no.1
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• pp.1-38
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• 1979

The woody species of Genus Ilex which are endemic to Korea are distributed on limited area due to solely temperature factor. There is some differences according to species, however in general, the evergreen Ilex are found along southern coastal area of Korean Peninsula and near islands where the cold index does not exceed $-5^{\circ}C$. But Ilex macropoda and the variety, only deciduous ones, are grown in temperate zone of the peninsula and some islands. The list of Ilex species of Korea are as follows. Ilex cornuta Lindley et Pax., I. crenata Thunb. var. microphylla Max., I. crenata Thunb., I. rotunda Thunb., I. macropoda Miq., I. macropoda Miq. var. pseudo-macropoda Loensner, I. integra Thunb. The author surveyed the populations of Ilex species as many as possible and data of some characters such as leaf shape, spine, fruit shape, stomata density, sex ratio in natural communities, etc. are collected. Almost all the Ilex species in Korea show sporadic distribution. This means quite small sized populations isolate distantly each other eliminating the change of gene exchange in between. Particularly Ilex conuta and I. crenata show the morphological differentiation among populations as well as significant individual variation within a population. These were true with such characteristics, leaf shape, leaf dimension, leaf margin, fruit shape, spine, and stomata density. The founded are that the fruit length and the stomata density counted on the beneath surface of leaves of Ilex cornuta increased with the decrease of latitude. These are naturally closely related with the cold index values. The table shown below indicates the correlation between mean stomata density per $0.3642mm^2$ and cold index values. These relation however were not observed on Ilex crenata. The most dominated natured in relation to individual variation were outline of leaf, the number of marginal spine, the shape of leaf cross section and the degree of luster of the upper leaf surface. As shown in photos 5~7, these variations are agreed at a glance. There are reports that the development of marginal spines in some Ilex species is associated with the juvenility and topophysis. In present study, these two factors were neglected because of the intended sampling procedure. Of Ilex rotunda, population difference with the characteristics of leaf length is recognized but not for leaf width, petiole length, and fruit size. However, individual variations within a population were significantly large. In case of Ilex integra, only individual differences within population were calculated statistically for such characteristics as leaf length, leaf width, and petiole length. As to natural population, the sex ratio was 1:2 (female to male) for Ilex cornuta, and 1:1 for Ilex crenata. The tendency of more male than female in I. cornuta was agreed to other observations. Preparing the tip cutting of length 10cm, and treating with IBA, then attaching earth ball to the cut end, very successful rooting percentages were obtained. Asexual propagation has the advantages of maintaining the heterozygosity of existing varieties and overcoming the difficulties of delayed seed germination frequently encountered with Ilex species. Considering a great deal of variation in morphological traits, a good possibility of selection breeding for decorative and ornamental purposes exists. At present, these evergreen Ilex are ignored by local people as nuisance weedy shrubs. So the proper protection measures should promptly be taken.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.4 no.1
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• pp.1-23
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• 1968

This study aimed at contributing to the improvement of cropping systems after finding out the effects of excrements and components of crop root influence on other crops as well as themselves. The following forage crops suitable for our country were selected for the present study. Aqueous extracts of fresh roots, aqueous extracts of rotting roots and aqueous solutions of excrements of red clover, orchard grass and brome grass were studied for the effects influencing the germination and growth of seedlings of red clover, ladino clover, lespedeza, soybean, orchard grass, Italian ryegrass, brome grass, barley, wheat, sorghum, corn and Hog-millet. In view of the possibility that the organic acid might be closely related to the excrements and components of crop root connected with soil sickness, the acid components of three species of roots were analysed by paper chromatography and gas chromatography method. The following results were obtained: 1. Effects of Aqueous Extracts of Fresh Roots : Aqueous extracts of red clover: The extracts inhibited the growth of seedlings of the ladino clover and lespedeza and also inhibited the development of most crops except that of sorghum among the Graminaceae. Aqueous extracts of orchard grass: The extracts promoted the seedlings growth of red clover and soybean, while it inhibited the germination and growth of orchard grass. There were no noticeable effects influencing other crops while it inhibited the growth of barley and Hog-millet. Aqueous extracts of brome grass: There was no effect on Italian ryegrass but there was an inhibiting effect on the other crops. 2. Effects of Aqueous Extracts of Rotting Roots : Aqueous extracts of red clover: The extracts promoted the seedling growth of red clover. But it reflected the inhibiting effects on other crops except sorghum. Aqueous extracts of orchard grass: The extracts promoted the growth of red clover, ladino clover, soybean and sorghun, while it inhibited the germination and rooting of barley and Hog-millet. Aqueous extracts of brome grass: The extracts gave the promotive effects to the growth of red clover, soybean and sorghum, but caused inhibiting effects on orchard grass, brome grass, barley and Hog-millet. 3. Effects of Aqueous Solutions of Excrements : The aqueous solution of excrements of red clover reflected the inhibition effects to the growth of Graminaceae, while the aqueous solutions of excrements of orchard grass and Italian ryegrass caused the promotive effects on the growth of red clover. 4. Results of Organic Acid Analysis : The oxalic acid, citric acid, tartaric acid, malonic acid, malic acid and succinic acid were included in the roots of red clover as unvolatile organic acid, and in the orchard grass and brome grass there were included the oxalic acid, citric acid, tartaric acid and malic acid. And formic acid was confirmed in the red clover, orchard grass and brome grass as volatile organic acid. In consideration of the results mentioned in above the effects of excrements and components of roots found in this studies may be summarized as follows. 1) The red clover generally gave a disadvantageous effect on the Graminaceae. Such trend was considered chiefly caused by the presence of many organic acids, namely oxalic, citric, tartaric, malonic, malic, succinic and formic acid. 2) The orchard grass generally gave an advantageous effect on the Leguminosae. This may be due to a few kinds of organic acid contained in the root, namely oxalic, citric, tartaric, malic and formic acid. Furthermore a certain of promotive materials for growth was noted. 3) As long as the root of brome grass are not rotten, it gave a disadvantageous effect on the Leguminosae and Graminaceae. This may be due to the fact that several unidentified volatile organic acid were also included besides the confirmed organic acid, namely oxalic, citric, tartaric, malic and formic acid. 5. Effects of Components in Roots to the Soil Sickness : 1) It was considered that the cause of alleged red clover's soil sickness did not result from the toxic components of the roots. 2) It was recognized that the toxic components of roots might be the cause of soil sickness in case the orchard grass and brome grass were put into the long-term single cropping. 6. Effects of Rooted Components to the Companion Crops in the Cropping System : a) In case of aqueous extracts of fresh roots and aqueous excrements (Inter cropping and mixed cropping) : 1) Advantageous combinations : Orchard grass->Red clover, Soybean, Italian ryegrass->Red clover, 2) Disadvantageous combinations : Red clover->Ladino clover, Lespedeza, Orchard grass, Italian ryegrass, Fescue Ky-31, Brome grass, Barley, Wheat, Corn and Hog.millet, Orchard grass->Lespedeza, Orchard grass, Barley and Hog-millet, Brome grass->Red clover, Ladino clover, Lespedeza, Soybean, Orchard grass, Brome grass, Barley, Wheat, Sorghum, Corn and Hog-millet, 3) Harmless combinations : Red clover->Red clover, Soybean and Sorghum, Orchard grass->Ladino clover, Italian ryegrass, Brome grass, Wheat, Sorghum and Corn, Brome grass->Italian ryegrass, b) In case of aquecus extracts of rotting roots(After cropping) : 1) Advantageous combinations : Red clover->Red clover and Sorghum, Orchard grass->Red clover, Ladino clover, Soybean, Sorghum, and Corn, Brome grass->Red clover, Soybean and Sorghum, 2) Disadvantageous combinations : Red clover->Lespedeza, Orchard grass, Italian ryegrass, Brome grass, Barley, Wheat, and Hog-millet Orchard grass->Barley and Hog-millet, Brome grass->Orchard grass, Brome grass, Barley and Hog-millet, 3) Harmless combinations : Red clover->Ladino clover, Soybean and Corn, Orchard grass->Lespedeza, Orchard grass, Italian ryegrass, Brome grass and Wheat Brome gass->Ladino clover, Lespedeza, Italian ryegrass and Wheat.

• Applied Biological Chemistry
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• v.9
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• pp.125-147
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• 1968

This experiment was carried out as a part of the studies on reasonable application of nitrogen in rice plant to determine: (I) Nitrogen absorption. and rooting of rice seedlings as affected by urea foliar application at late seedling stage (II) Effect of leaf prunning and foliar application of urea at late heading stage on the maturation and yield of rice (III) Effect of foliar application of urea and its time during the stage of ear formation on yield of rice plant. Results obtained are summarized as follows. Exp.I: Nitrogen absorption and rooting of rice seedlings as affected be urea foliar application at late seedling stage. 1 : The foliar application of urea plots$(T_{1},T_2)$ snowed mare N-content than non-urea foliar application plot(T0) at lane seedling stage, being significant among treatments and foliar application of urea seemed more effective in increasing the N-content of seedlings. and promoted root settlement and early growth alter the transplanting. 2 : The carbon contents of the plants of $T_1$, and $T_2$ at late seedling stage increased than T0, and the carbon contents. of $T_1$ and $T_2$ plots became higher in amount in proportion to the nitrogen absorption as compared with those of $T_0$. 3 : C/N ratio appeared significant among soil application plots($N_1, \;N_2$) and foliar application of urea plots ($T_1$, $T_2$ and $T_0$). C/N ratio was lower in case of increased amount of nitrogen. The higher contents of nitrogen and carbon and lower C/N ratio resulted in the increment of root numbers and root lengths. Exp.II: Effect of leaf prunning and foliar application of urea at late heading stage on the maturation and yield of rice. 1 : There was a highly significant decrease in the maturing rate by severe leaf prunning. In the mean time, significant increase in maturing rate was observed with urea foliar application and it was found the more frequent application the more effective for higher maturing rate with a moderate significance. A correlationship between the level of prunning and maturing rate was enumerated to 0.961 of correlation coefficient, which indicated an increased maturing rate by the increased number of remaining leaves. 2 : The 1.000 grain weight, grain weight and hulled rice yield increased by leaf prunning in order (plot a$A_1$, $A_3$, $A_2$ and $A_0$ were 89.8%, 89.4%, 87.8% and 87.5% respectively, showing the highest of rate in $A_1$ and $A_3$ in methods of ear fertilization and being highly significant between its treatment. 3 : 1000 grain weights were highly significant between time of application, showing a tendency of increase of weights with the time lagging until days before earings as that of maturing rates. High significance was recognized between methods of ear fertilization, showing the highest in $A_2$ 23.18 gr. 4 : Yields per $3.3m^2$ were not significant between time of ear fertilization, whereas were highly significant between methods of ear fertilization. Those of $A_1$, $A_3$, $A_2$ and $A_0$ were 1.486 kg, 1.491 kg, 1.381 kg and 1.328 kg, respectively, showing the highest in $A_1$ and $A_3$. 5 : Hulling ratios showed significant different between time of ear fertilization, showing the highest in $T_2$, whereas those of methods of ear fertilization were highly significant between its treatment, Those of $A_1$, $A_3$, $A_2$ and $A_0$ were 84.72%, 84.06%, 83.29%, and 82.56% respectively, showing the highest m $A_2$ and $A_3$ among others. 6 : Yields of hulled rice per $3.3m^2$ showed significant different between time of ear fertilization, showing the highest in $T_1$ 1.192 kg. Whereas, those were highly significant between methods of ear fertilization. Those of $A_1$, $A_3$, $A_2$ and $A_0$ were 1.259 kg, 1.254 kg, 1.149 kg and 1.095 kg, respectively, showing the highest in $A_1$ and $A_2$. 7 : Contents of nitrogen on rice plant increased in case of nitrogen application as ear fertilizer and showed that the case of urea foliar application was more effective than that of soil application, showing the increased nitrogen content of rice plant was accompanied by carbon content.