• 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.22 no.1
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• pp.135-166
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• 1977

Some biochemical features of varietal variation in seed protein and their implications for soybean breeding for high protein were pursued employing 86 soybean varieties of Korea, Japan, and the U.S.A. origins. Also, studied comparatively was the temporal pattern of protein components accumulation during seed development characteristic to the high protein variety. Seed protein content of the 86 soybean varieties varied 34.4 to 50.6%. Non-existence of variety having high content of both protein and oil, or high protein content with average oil content as well as high negative correlation between the content of protein and oil (r=-0.73$^{**}$) indicate strongly a great difficulty to breed high protein variety while conserving oil content. The total content of essential amino acids varied 32.82 to 36.63% and the total content of sulfur-containing amino acids varied 2.09 to 2.73% as tested for 12 varieties differing protein content from 40.0 to 50.6%. The content of methionine was positively correlated with the content of glutamic acid, which was the major amino acid (18.5%) in seed protein of soybean. In particular, the varieties Bongeui and Saikai ＃20 had high protein content as well as high content of sulfur-containing amino acids. The content of lysine was negatively correlated with that of isoleucine, but positively correlated with protein content. The content of alanine, valine or leucine was correlated positively with oil content. The seed protein of soybean was built with 12 to 16 components depending on variety as revealed on disc acrylamide gel electrophoresis. The 86 varieties were classified into 11 groups of characteristic electrophoretic pattern. The protein component of Rm=0.14(b) showed the greatest varietal variation among the components in their relative contents, and negative correlation with the content of the other components, while the protein component of Rm=0.06(a) had a significant, positive correlation with protein content. There was sequential phases of rapid decrease, slow increase and stay in the protein content during seed development. Shorter period and lower rate of decrease followed by longer period and higher rate of increase in protein content during seed development was of characteristic to high protein variety together with earlier and continuous development at higher rate of the protein component a. Considering the extremely low methionine content of the protein component a, breeding for high protein content may result in lower quality of soybean protein.n.