• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.105-110
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• 1995

The experiment were carried out to obtain a somatic hybrid through protoplast fusion between P.hybrid and N. sanderae. The isoenzyme pattern he chromosome number and the phonotype were observed for genetic study on the regenerants obtained from the fusion product cultures. Putative somatic hybrids possessed all the bands that appeared in both mother plane. A specific band was found on the top of the banding pattern which was assumed to be a marker band of somatic hybrid between two genera. Aspartate amninotransferase isoenzyme bands which were found in both mother plane were also revealed in the putative somatic hybrids or deleted in the upper part of H. sanderae band pattern. The chromosome number of P.hybrida was 2n=14, while N, sanderae was 2n=18,but the number of the putative somatic hybrids ranged from n=32 to 36. The phonotype of putative somatic hybrids was intermediate of the mother plants.

• Current Research on Agriculture and Life Sciences
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• v.10
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• pp.147-155
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• 1992

This study was conducted to get basic information for factors affecting protoplast fusion between Petunia hybrida 'Titan red' and Nicotiana sanderae. The experiments such as fusogen, time of PEG treatment, temperature at fusion, $CaCl_2{\cdot}2H_2O$ concentration in fusion solution, and $CaCl_2{\cdot}2H_2O$ concentration and pH in eluting solution were carried out to increase the fusion efficiency. The results obtained were as follows; Fusion between P. hybrida and N. sanderae was accelerated when the mixture of the protoplasts was treated with 30% PEG 6,000 solution containing 5.5 mM $CaCl_2{\cdot}2H_2O$ for 10 minutes at $25^{\circ}C$, and subsequently eluted with a eluting solution containing 50 mM $CaCl_2{\cdot}2H_2O$ adjusted to pH 9.0.

• Journal of the Korean Society of Tobacco Science
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• v.11 no.2
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• pp.127-134
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• 1989

Several growth characteristics of two ornamental tobacco species, Nicotiana sanderae and N. affinis, were investigated in this study. Also effect of temperature and daylength on the flowering of the tobacco plants were evaluated to obtain basic information on breeding and cultivation. 1. The plants were great in high temperature-long day at the early stage and in low temperature-short day at the late stage of plant growth, for both Nicotana species. At the early growth stage the leaf length N. sanderae was great in high temperature-long day, and that of N. affinis was great in high temperature-short day period, while at the late stage of the plant growth the leaf lengths were more significantly effected by the temperature rather than daylength. Leaf width and leaf shape index were less sensitive to the conditions. 2. For both of the species, the total number of tobacco leaves not much influenced by the temperature and daylength. 3. There were no significant differences for budding and flowering period between the two species, both of which were sensitive to temperature and daylength with more influence by daylength than temperature. 4. Number of floral stalks, number of flower and flowering period were not much influenced by temperature and daylength; however, N. affinis had 2 more floral stalks, 31 more flowers, and 6 day longer flowering period than N. sanderae.

• Applied Biological Chemistry
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• v.37 no.5
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• pp.343-348
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• 1994

Ti plasmid of A. tumefaciens was labeled with $^3H-thymidine$, purified and encapsulated into phosphatidylserine (PS) and PS-cholesterol (Chol; 1 : 1 molar ratio) liposomes by lyophilization-rehydration method. PS was supplemented with 1 mole percent octadecyl rhodamine B for fluorometric measurement of PS. Liposomes entrapping $^3H-Ti plasmid$ were fused with Nicotiana sanderae protoplasts by treating with 5 mM $CaCl_2$ and 10% PEG. The fusion was evidenced by fluorescence microscopic technique. The amounts of Ti plasmid and PS associated with protoplasts were assayed by the radioactivity of $^3H-Ti plasmid$ and by the fluorescence of rhodamine B. About 7.9% of the PS liposome and 7.2% of PS-Chol liposome were fused with protoplasts. During the fusion process, about 30% of the liposomal contents of PS-Chol liposome was leaked, in contrast to about 60% leakage of its contents in PS liposome. Accounting the number of liposomes fused with protoplasts together with the encapsulation efficiency and the leakage of liposomal contents, it was calculated that ca. 1,700 Ti plasmid was transfered into one protoplast by the present method. This result may indicates that the present method transfers enough Ti plasmid into plant protoplast to elicit genetic transformation of plants.