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A study was initiated to characterize key enzymes that influence sweetness in carrot (Daucus carota L.) roots. Sucrose synthase (SS), sucrose phosphate synthase (SPS), and UDP-glucose pyrophosphorylase (UDPL) genes were isolated from potato (Solanum tuberosum L.) and cloned in an anti-sense orientation into Agrobacterium tumefaciens Bin19, which has a CaMV 35S promoter. Seedling hypocotyl sections of selected carrot lines were pre-incubated on B5 medium for 2 days, co-cultivated with A. tumefaciens Bin 19 for additional 3 days, and then transferred to a modified B5 medium containing 50 g/mL kanamycin and 400 g/mL carbenicillin. In 4 weeks, 18.6%, 33.3%, and 26.7% of the cultures from a breeding line (W204-C) were found to be transformed, respectively, with SS, SPS, and UDPL as determined by kanamycin resistance. In contrast, no kanamycin-resistant calli were obtained from a commercial cultivar (Navajo) in these transformation studies. The transformed calli proliferated in the medium containing 50 g/mL kanamycin and 400 g/mL carbenicillin, whereas non-transformed calli died in the same medium. These transformed calli are currently being used to regenerate plants via asexual embryogenesis using a suspension culture. The influence of these additional genes on sugar metabolism and accumulation in root tissues of transformed carrots will be characterized in the future.
The phenotypic expression and inheritance of the rolC gene in the transgenic plants of Salpiglossis sinuata L. were investigated. The chasmogamous salpiglossis plants with solid yellow flower color (ccrrDD) were transformed with Agrobacterium tumefaciens strains LBA4404 and EHA101 carrying rolC, GUS, and NPTII genes via a leaf disc co-cultivation system. The transgenic plants were shorter in plant height, produced more branches with a compact growth habit, and developed smaller flowers and narrower leaves as compared to the control plant. While the transgenic plants showed the same corolla color and color shades as the parental line, they became male sterile. A backcross between a male-sterile transgenic plant (ccrrDD plus rolC) and a nontransformed red-flowering line (ccRRDD) produced a progeny with red flower color and the same altered growth habit as the transgenic female parent. Only 4 out of 32 plants in this progeny population showed the negative GUS staining as well as the non transgenic phenotype. These results suggest that at least two copies of the rolC gene were integrated into one homologous chromosome pair during transformation and that a cross-over event may have occurred during meiosis.
The relative concentrations of sucrose, glucose, and starch in the xylem and cortex tissues of carrot (Daucus carota) roots were evaluated after harvest and during storage. For the three cultivars (Apache, Bolero, Danvers 126) tested, the cortex tissue contained 76.6, 49.1, and 33.6 mg·g–1 dry weight of sucrose, glucose, and starch, respectively. In comparison, the average contents of sucrose, glucose, and starch in xylem tissues were 57.4, 52.4, and 11.6 mg·g–1 dry weight, respectively. In general, cortex tissue contained higher concentrations of sucrose and starch than the xylem tissues. The glucose concentrations in cortex and xylem were similar. In `Apache', for example, the cortex tissue contained 40% and 57% higher concentrations of sucrose and starch, respectively, than the xylem tissues, whereas glucose content of the cortex was only 7.5% higher than that of the xylem. Since sweetness is largely influenced by sucrose, the relative volume of cortex to xylem must be considered in evaluating carrot cultivars for sweet taste.