, respectively. We decided partial amino acid sequences of P5CR-1 which showed the 70 to 80% homology to the deduced amino acid sequences of several plant P5CR cDNAs. Both isoenzymes had much lower affinity for NADH than for NADPH and were inhibited by free ATP and Mg 2+ ion. The inhibition was partially mitigated when ATP and Mg 2+ were added simultaneously to the reaction mixture. Cations at high concentration were inhibitory to P5CR activity. Interestingly, P5CR-2 was more stable to heat treatment at 40°°°°C than P5CR-1.

A method for Agrobacterium-mediated transformation of Freesia×hybrida is described. Cormlet-derived calli of two cultivars, 'Mosera' and 'Ishikawa f3' were co-cultivated with Agrobacterium tumefaciens strain EHA105 harboring the binary vector pIG121-Hm, which included hygromycin phosphotransferase gene and an intron-containing β-glucuronidase gene in the T-DNA region. Callus pieces were co-cultivated with A. tumefaciens on the callus proliferation medium [Murashige and Skoog (MS) medium containing 1 mg l −1 thidiazuron, 1 mg l −1 dicamba, 20 mg l −1 3′,5′-dimethoxy-4′-hydroxyacetophenone, 1% (w/v) glucose, 3% (w/v) sucrose, and 0.2% (w/v) Gelrite]. Then, they were cultured on the callus proliferation medium containing 300 mg l −1 cefatoxime and 10 mg l −1 hygromycin B. Hygromycin-resistant lines of both cultivars regenerated into plantlets after transfer onto MS medium containing 2 mg l −1 3-indoleacetic acid and 3 mg l −1 6-benzyl aminopurine and/or plant growth regulator-free MS medium. Transgenic plants were identified by β-glucuronidase assay and verified by Southern blot analysis. Two transgenic plant lines were obtained from 475 callus pieces of 'Mosera' , and one transgenic plant line was obtained from 290 callus pieces of 'Ishikawa f3' . This is the first report of the genetic transformation of Freesia. This method will allow the genetic improvement of this horticulturally important flower.

Breeding of a New Freesia Cultivar 'Ishikawa f2 go' (Freesia spp.

Crocetin glycosides such as crocin are noted as functional food materials since the preventive effects of crocin have been reported against chronic disease and cancer. However, it is unclear how these apocarotenoids are structurally changed through cooking for our intake. We examined such changes in crocetin glycosides (crocin, tricrocin, and crocin-3) contained in saffron (stigmas of Crocus sativus) through cooking models. These glycosides were almost kept stable in boiling for 20 min (a boiled cooking model), while hydrolysis of the ester linkage between glucose and the crocetin aglycone occurred in a grilled cooking model (180°C, 5 min), along with a 13-cis isomerization reaction in a part of crocetin subsequently generated. We further here revealed that the yellow petals of freesia (Freesia x hybrida) with yellow flowers accumulate two unique crocetin glycosides, which were identified to be crocetin (mono)neapolitanosyl ester and crocetin dineapolitanosyl ester. A similar result as above was obtained on their changes through the cooking models. Utility applications of the freesia flowers as edible flowers are also suggested in this study. Additionally, we evaluated singlet oxygen (1O2)-quenching activities of the crocetin glycosides contained in saffron and freesia, and crocetin and 13-cis crocetin contained in the grilled saffron, indicating that they possessed moderate 1O2-quenching activities (IC50 24–64 μM).

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