著者

Nazmul Hasan Naoki Tokuhara Takayuki Noda Nobuhiro Kotoda

出版者

Japanese Society for Plant Biotechnology

雑誌

Plant Biotechnology (ISSN:13424580)

巻号頁・発行日

vol.40, no.1, pp.51-62, 2023-03-25 (Released:2023-03-25)

参考文献数

66

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Shortening the juvenility is a burning issue in breeding fruit trees such as Satsuma mandarin (Citrus unshiu Marc.). Decreasing the breeding period requires a comprehensive understanding of the flowering process in woody plants. Throughout the Arabidopsis flowering system, FLOWERING LOCUS T (FT) interacts with other transcription factors (TFs) and functions as a transmissible floral inducer. In a previous study, a VASCULAR PLANT ONE-ZINC FINGER1 (VOZ1)-like TF from the Satsuma mandarin, CuVOZ1, showed protein–protein interaction with two citrus FTs in a yeast two-hybrid (Y2H) system and precocious flowering in Arabidopsis. In this study, another VOZ, CuVOZ2, was isolated from the Satsuma mandarin ‘Aoshima’ and protein–protein interaction was confirmed between CuVOZ2 and CuFTs. No apical meristem (NAM) and zinc coordination motifs were identified within the N-terminal of CuVOZ2. Docking simulation predicted that interactions between CuVOZ2 and CuFTs might occur in domain B of CuVOZ2, which contains a zinc finger motif. According to docking predictions, the distances between the amino acid residues involved ranged from 1.09 to 4.37 Å, indicating weak Van der Waals forces in the interaction. Cys216, Cys221, Cys235, and His239 in CuVOZ2 were suggested to bond with a Zn2+ in the Zn coordination motif. Ectopic expression of 35SΩ:CuVOZ2 in Arabidopsis affected the flowering time, length of inflorescence and internode, and number of siliques, suggesting that CuVOZ2 might regulate both vegetative and reproductive development, act as a trigger for early flowering, and be involved in the elongation of inflorescence possibly in a slightly different way than CuVOZ1.

著者

Nobuhiro Kotoda Satoshi Matsuo Ichiro Honda Kanako Yano Tokurou Shimizu

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-016, (Released:2016-09-27)

被引用文献数

5

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Gibberellins (GAs) are involved in many aspects of plant growth and development in higher plants. In this study, we aimed to characterize Satsuma mandarin (Citrus unshiu Marc.) GA 2-oxidase genes encoding enzymes with GA inactivation activity because the accumulation of active GAs is regulated by the balance between their synthesis and inactivation. We showed that CuGA2ox4, CuGA2ox2/3, and CuGA2ox8 were differentially expressed in various tissues in Satsuma mandarin and that these genes functioned like GA 2-oxidase genes in transgenic Arabidopsis. The expression pattern of CuGA2ox4 resembled that of CuGA20ox1, which was reported previously as a Satsuma mandarin GA 20-oxidase gene, in the reproductive tissues of the adult trees in that both genes were expressed in juice sacs in November and December and peel in December, suggesting that CuGA2ox4 and CuGA20ox1 act in concert to regulate the accumulation of active GAs in fruit at the maturing stage. On the other hand, CuGA2ox2/3 and CuGA2ox8 were more highly expressed in seeds and flower buds, respectively. Further study of GA biosynthetic genes, including GA 2-oxidase genes, would provide insight into the mechanism of flowering, fruit development, seedlessness, biennial bearing, and peel puffins of citrus such as Satsuma mandarin.

著者

Nobuhiro Kotoda Satoshi Matsuo Ichiro Honda Kanako Yano Tokurou Shimizu

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.MI-085, (Released:2015-11-28)

被引用文献数

14

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Satsuma mandarin (Citrus unshiu Marc.) has two GA 20-oxidase genes, CuGA20ox1 and CuGA20ox2, and the genomic sequence for CuGA20ox1 is shorter than that for CuGA20ox2, although the coding region of cDNA for CuGA20ox1 is slightly longer than that for CuGA20ox2. Southern blot analysis revealed that 12 Citrus cultivars examined and trifoliate orange [Poncirus trifoliata (L.) Raf.] have at least two types of GA 20-oxidase genes, such as CuGA20ox1 and CuGA20ox2 genes. CuGA20ox1 and CuGA20ox2 were differentially expressed in various tissues. CuGA20ox1 was expressed in almost all of the tissues investigated with relatively higher expression in vegetative than in reproductive tissues, whereas CuGA20ox2 was specifically expressed in flower buds just before anthesis. These distinct expression patterns of CuGA20ox1 and CuGA20ox2 imply that function of these two genes diverged in the process of evolution. The specific and relatively higher expression of CuGA20ox2 in flower buds would explain why GA-like activity was higher in Satsuma mandarin’s ovaries at anthesis. Transgenic Arabidopsis [Arabidopsis thaliana (L.) Heynh] plants ectopically expressing CuGA20ox1 or CuGA20ox2 were examined to elucidate the function of these two Satsuma mandarin genes. Phenotypic analysis revealed that both CuGA20ox1 and CuGA20ox2 caused elongated inflorescence but did not affect the timing of flowering in transgenic Arabidopsis as compared with wild-type controls. Ectopic expression of CuGA20ox1 and CuGA20ox2 significantly affected the levels of GA24 and GA34 on the non-13-hydroxylation pathway; GA24 decreased and GA34 increased. This observation indicates that both CuGA20ox1 and CuGA20ox2 accelerated the conversion of GA24, a substrate of a GA 20-oxidase, to GA9, a precursor of an active form of GA4. Likewise, on the early-13-hydroxylation pathway, ectopic expression of CuGA20ox1 significantly decreased GA19 and increased GA29 and GA8, inactive metabolites of 2-hydroxylation of GA20 and GA1, respectively, suggesting the activation of this biosynthetic pathway. CuGA20ox2 also had a tendency to activate the early-13-hydroxylation pathway although it increased only GA20 with a statistically significant difference. Taken together, we concluded that CuGA20ox1 and/or CuGA20ox2 activated both the early-13- and non-13-hydroxylation pathways for increasing active GAs, resulting in elongated inflorescences in transgenic Arabidopsis.