著者

Yoshiki Tanahara Kaho Yamanaka Kentaro Kawai Yukiko Ando Takashi Nakatsuka

出版者

Japanese Society for Plant Biotechnology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.39, no.3, pp.273-280, 2022-09-25 (Released:2022-09-25)

参考文献数

28

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Matthiola incana is an important floricultural plant that blooms from winter to spring, and had been desired to be established a transformation system. This study successfully obtained stable transgenic plants from M. incana. We used Agrobacterium tumefaciens harboring a binary vector containing the β-glucuronidase gene (GUS) under the control of cauliflower mosaic virus 35S promoter to evaluate the transformation frequency of M. incana. We observed that cocultivation with the A. tumefaciens strain GV3101 for 5 days effectively enhanced the infection frequency, assessed through a transient GUS expression area in the seedling. Furthermore, the addition of 100 µM acetosyringone was necessary for Agrobacterium infection. However, we could not obtain transgenic plants on a shoot formation medium supplemented with 1 mg l−1 6-benzyladenine (BA). For callus formation from the leaf sections, a medium supplemented with 1–50 µM fipexide (FPX), a novel callus induction chemical, was employed. Then, the callus formation was observed after 2 weeks, and an earlier response was detected than that in the BA medium (4–6 weeks). Results also showed that cultivation in a selection medium supplemented with 12.5 µM FPX obtained hygromycin-resistant calli. Thus, this protocol achieved a 0.7% transformation frequency. Similarly, progenies from one transgenic line were observed on the basis of GUS stains on their leaves, revealing that the transgenes were also inherited stably. Hence, FPX is considered a breakthrough for establishing the transformation protocol of M. incana, and its use is proposed in recalcitrant plants.

著者

Eri Tomizawa Shogo Ohtomo Kanako Asai Yuka Ohta Yukako Takiue Akihiro Hasumi Masahiro Nishihara Takashi Nakatsuka

出版者

Japanese Society for Plant Biotechnology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.38, no.3, pp.323-330, 2021-09-25 (Released:2021-09-25)

参考文献数

31

被引用文献数

6

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Betalains, comprising violet betacyanins and yellow betaxanthins, are pigments found in plants belonging to the order Caryophyllales. In this study, we induced the accumulation of betalains in ornamental lisianthus (Eustoma grandiflorum) by genetic engineering. Three betalain biosynthetic genes encoding CYP76AD1, dihydroxyphenylalanine (DOPA) 4,5-dioxygenase (DOD), and cyclo-DOPA 5-O-glucosyltransferase (5GT) were expressed under the control of the cauliflower mosaic virus (CaMV) 35S promoter in lisianthus, in which anthocyanin pigments are responsible for the pink flower color. During the selection process on hygromycin-containing media, some shoots with red leaves were obtained. However, most red-colored shoots were suppressed root induction and incapable of further growth. Only clone #1 successfully acclimatized and bloomed, producing pinkish-red flowers, with a slightly greater intensity of red color than that in wild-type flowers. T1 plants derived from clone #1 segregated into five typical flower color phenotypes: wine red, bright pink, pale pink, pale yellow, and salmon pink. Among these, line #1-1 showed high expression levels of all three transgenes and exhibited a novel wine-red flower color. In the flower petals of line #1-1, abundant betacyanins and low-level betaxanthins were coexistent with anthocyanins. In other lines, differences in the relative accumulation of betalain and anthocyanin pigments resulted in flower color variations, as described above. Thus, this study is the first to successfully produce novel flower color varieties in ornamental plants by controlling betalain accumulation through genetic engineering.

著者

Masumi Yamagishi Takashi Nakatsuka

出版者

一般社団法人 園芸学会

雑誌

The Horticulture Journal (ISSN:21890102)

巻号頁・発行日

pp.OKD-057, (Released:2017-02-28)

被引用文献数

14

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Genes encoding a MYB12 transcription factor, which regulates anthocyanin biosynthesis, are the key genes causing variations in the anthocyanin colors in lily flowers. However, the origin of the MYB12 in Asiatic hybrid lilies (Lilium spp.) is not completely known. In this study, we analyzed anthocyanin pigments in tepals of L. maculatum, L. lancifolium, L. callosum, L. leichtlinii, L. davidii, L. bulbiferum, and L. dauricum, and clarified that L. dauricum and L. bulbiferum accumulated a single anthocyanin, cyanidin 3-O-rutinoside, in entire tepals, although these wild species had orange-colored tepals. The sequencing of MYB12 genes revealed seven allelic sequences among ten L. dauricum plants and two allelic sequences in one L. bulbiferum plant. These MYB12 sequences were the same as or similar to the sequences isolated from pink Asiatic hybrid lily cultivars, indicating that L. dauricum and L. bulbiferum have contributed to the anthocyanin coloration of these cultivars.