Transgenic lines of creeping bent grass were generated by Agrobacterium-mediated transformation with the VuNCED1 which was cloned from cow pea has a homology to 9-cis-epoxycarotenoid dioxygenase, which is supposed to be involved in abscisic acid (ABA) biosynthesis. ABA, a cleavage product of carotenoids, is involved in stress responses in plants. The limiting step of ABA biosynthesis in plants is presumably the cleavage of 9-cis-epoxycarotenoids, the first committed step of ABA biosynthesis. Molecular analyses of transgenic lines as performed by Southern hybridization genomic DNA-PCR revealed integration of the VuNCED1. Challenge studies performed with transgenic plants by exposure to salt stress (up to 10 dS m À1 ) and water stress (up to 75%) for 10 weeks, revealed that more than 50% of the transgenic plants could survive NaCl and drought stress whereas wild-type was not. ABA levels were measured under drought and normal conditions, endogenous ABA was dramatically increased by drought and NaCl stress in transgenic plants. These results indicate that it is possible to manipulate ABA levels in plants by over expressing the key regulatory gene in ABA biosynthesis and that stress tolerance can be improved by increasing ABA levels.

A new selection system for onion transformation that does not require the use of antibiotics or herbicides was developed. The selection system used the Escherichia coli gene that encodes phosphomannose isomerase (pmi). Transgenic plants carrying the manA gene that codes for pmi can detoxify mannose-6-phosphate by conversion to fructose-6-phosphate, an intermediate of glycolysis, via the pmi activity. Six-week-old embryogenic callus initiated from seedling radicle was used for transformation. Transgenic plants were produced efficiently with transformation rates of 27 and 23% using Agrobacterium and biolistic system, respectively. Untransformed shoots were eliminated by a stepwise increase from 10 g l(-1) sucrose with 10 g l(-1) mannose in the first selection to only 10 g l(-1) mannose in the second selection. Integrative transformation was confirmed by PCR, RT-PCR and Southern hybridization.

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Shoot-tip culture was used to produce clonal plants of radish stock seeds. Using 6-benzyladenine (BA), the largest number of RA2 line multi-shoots were formed with an average of 14.67 shoots on 1.33 µM BA in early seedlings and 11.33 shoots on 1.78 µM BA in juvenile seedlings. The largest number of RA4 line multi-shoots were formed with an average of 11.67 shoots on 2.22 µM BA in early seedlings and 13.67 shoots on 1.33 µM BA in juvenile seedlings. There was little difference in the significance level by BA concentration in both lines. Using Thidiazuron (TDZ), the number of RA2 line multi-shoots increased with increasing TDZ concentration, forming the largest number of multi-shoots in 0.45 µM TDZ (7.0 and 3.0 multi-shoots for early and juvenile seedlings, respectively), but few multi-shoots were formed from TDZ 2.25 and 4.5 µM. RA4 line produced almost no multi-shoots in early seedlings, and 3.7 multi-shoots were produced in 0.23 and 0.45 µM TDZ in juvenile seedlings, but not at higher concentrations. Analysis of the tissue culture seedlings grown by cultivating the generated multi-shoots with Radish Foundation seeds using SSR marker revealed a weak pattern of mutation in the generated tissue culture seedlings, but there was no mutant. In addition, in terms of root roots, both RA2 and RA4 lines generally had the best rooting, number of roots, and degree of root development in 4.9 µM indol-3-butyric acid (IBA).