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Tsuyama, Japan

Nakamura N.,Suntory Holdings Ltd | Tems U.,Florigene Pty Ltd | Fukuchi-Mizutani M.,Suntory Holdings Ltd | Chandler S.,Florigene Pty Ltd | And 4 more authors.
Plant Biotechnology | Year: 2011

An important part of the assessment of the potential environmental impact from the introduction of a genetically modified (GM) plant is an evaluation of the potential for gene flow from the GM plant to related wild species. This information is needed as part of the risk-assessment process, in the context of whether gene flow to wild species is possible. One method for evaluating gene flow is to use molecular techniques to identify genes in wild species populations that may have originated from a cultivated species. An advantage of this method is that a phenotypic marker or trait is not required to measure gene flow. In the present study we analyzed the seedlings of seeds from three wild native Rosa species (R. multiflora Thunb., R. luciae Rochebr. et Franch. ex Crép. and R. rugosa Thunb.) selected from several locations across Japan where the wild rose was growing in close proximity to cultivated rose plants (Rosxhybrida). To determine whether gene flow from cultivated rose had occurred, young leaves of 1,296 seedlings from the wild Rosa plants were analyzed by PCR for the presence of the KSN locus. This locus originated from a sport of R. chinensis Jacq. var. spontanea (Rehd. Et Wils.) Yu et Ku and is involved in the recurrent flowering phenotype observed for cultivated rose hybrids, but is absent in Japanese species roses. The KSN locus was absent in all seedlings sampled, indicating no gene flow to wild Rosa species from the cultivated rose had occurred, and providing evidence that the probability of gene flow from cultivated to wild Rosa species in Japan is low or non-existent.

Okayama T.,NISSHOKU Corporation | Furukawa H.,Osaka Prefecture University | Okamura K.,NISSHOKU Corporation | Murase H.,Osaka Prefecture University
Environmental Control in Biology | Year: 2010

We investigated the effects of different photoperiods on the expression level of GUS genes regulated by the CaMV-35S promoter in leaf lettuce (Lactuca sativa L. cv. 'Greenwave') for a recombinant protein production system. A leaf disk experiment and a cultivation experiment were conducted in different photoperiods. A linear relationship (R2 = 0.9934) was observed between the lengths of photoperiods and GUS activities per unit fresh weight in the leaf disk experiment. This result indicates that the expression of the CaMV-35S promoter during a photoperiod was much more enhanced than that during a dark period. In the cultivation experiment, a longer photoperiod helped not only to enhance the growth of a lettuce plant but also to increase its GUS activity. As a result, GUS activity per plant under the 24:0 h photoperiod was significantly (P<0.05) higher than that under the 16:8 and 8:16 h photoperiods. However, the question remains whether these results for GUS-CaMV-35S promoter can be applied to other recombinant proteins or promoters. Further studies are thus required to establish a practical strategy for designing an efficient recombinant production system, in a plant factory.

Nakamura N.,Suntory Holdings Ltd | Fukuchi-Mizutani M.,Suntory Holdings Ltd | Katsumoto Y.,Suntory Holdings Ltd | Togami J.,Suntory Holdings Ltd | And 12 more authors.
Plant Biotechnology | Year: 2011

The release of genetically modified plants into the environment can only occur after permission is obtained from the relevant regulatory authorities. This permission will only be obtained after extensive risk assessment shows comparable risk of impact to the environment and biodiversity as compared to non-transgenic host plants. Two transgenic rose (Rosa×hybrida) lines, whose flowers were modified to a bluer colour as a result of accumulation of delphinidin-based anthocyanins, have been trialed in greenhouses and the field in both Japan and Australia. Flower colour modification was due to expression of genes of a viola flavonoid 3',5'-hydroxylase and a torenia anthocyanin 5-acyltransferase. In all trials it was shown that the performance of the two transgenic lines, as measured by their growth characters, was comparable to the host untransformed variety. Biological assay showed that the transgenic lines did not produce allelopathic compounds. In Japan, seeds from wild rose species that had grown in close proximity to the transgenic roses did not carry either a Rosa×hybrida specific marker gene or the transgenes. In hybridization experiments using transgenic rose pollen and wild rose female parents, the transgenes were not detected in the seed obtained, though there was a low frequency of seed set. The transgene was also not transmitted when Rosa×hybrida cultivars were used as females. In in situ hybridization analysis transgene transcripts were only detected in the epidermal cells in the petals of the transgenic roses. In combination, the breeding and in situ analysis results show that the transgenic roses contain the transgene only in the L1 layer cells and not in the L2 layer cells that generate reproductive cells. General release permissions have been granted for both transgenic lines in Japan and one is now commercially produced.

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