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Dineshkumar Selvaraj, Sherif Sherif, Mohd Sabri Pak Dek, Gopinadhan Paliyath, Islam El-Sharkawy, and Jayasankar Subramanian

downregulated. Our results suggest that CHS may be a limiting step in the production of anthocyanins in the european plum. Among the two copies of genes for UDP-glucose:flavonoid 3-O-glucosyltransferase, Pd-UFGT 1 expression levels were high in yellow 2 and

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Tomomi Tsuda, Masami Yamaguchi, Chikako Honda, and Takaya Moriguchi

We used RNA blot analysis to examine the expression of six genes of the anthocyanin biosynthesis pathway in the flowers and fruit skins at three developmental stages of white and red peaches and a deep-red nectarine [Prunus persica (L.) Batch]. In the red peach `Akatsuki' and the deep-red nectarine `Flavortop', expression levels of anthocyanin biosynthesis genes were related to anthocyanin accumulation in the fruit skin; expression of all six genes dramatically increased at Stage III of fruit development, and anthocyanin concentration also increased at this stage. In the white peach `Mochizuki', however, expression of the chalcone synthase gene (CHS) and the dihydroflavonol 4-reductase gene (DFR) was undetectable in Stage III, although the chalcone isomerase gene (CHI), the flavanone 3-hydroxylase gene (F3H), the anthocyanidin synthase gene (ANS), and the UDP-glucose-flavonoid 3-O-glucosyltransferase gene (UFGT) were expressed. We occasionally found red pigment in the skin of `Mochizuki' peach. In these red skin areas, both CHS and DFR were clearly expressed in Stage III. These results suggest that CHS and DFR are the key regulatory genes in the process of anthocyanin biosynthesis in mature red peach and nectarine.

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Paola S. Cotroneo, Maria P. Russo, Manuela Ciuni, Giuseppe Reforgiato Recupero, and Angela R. Lo Piero

Genes encoding chalcone synthase (CHS), anthocyanidin synthase (ANS), and UDP-glucose-flavonoid 3-O-glucosyltransferase (UFGT), some of the enzymes of anthocyanin biosynthetic pathway, were assayed in two different experiments using quantitative real-time reverse transcriptase (RT)-PCR, in order to test their transcription levels in the flesh of blood and common orange [Citrus sinensis (L.) Osbeck] fruit, and to investigate their role in anthocyanin accumulation in the same tissue. The first experiment compared a blood orange and a common orange cultivar during seven different fruit maturation stages. This was followed by the test of 11 different genotypes at the end of the winter season. Data collected from the first experiment, over the blood orange cultivar, were statistically analyzed using the Pearson correlation coefficient. Results show that CHS, ANS, and UFGT mRNA transcripts are up- and co-regulated in the blood orange cultivar, whereas they are down-regulated in the common orange cultivar. There is evidence of correspondence between the target genes expression level and the content of the pigment assessed. The second test confirms this correlation and shows that enzyme synthesis levels and pigment accumulation, in plants grown under the same environmental conditions, are dependent on the differences occurring among the genotypes tested. These results suggest that the absence of pigment in the common orange cultivars may be caused by the lack of induction on the structural genes expression. This is the first report on the characterization of the relationships between biosynthetic genes expression and fruit flesh anthocyanin content in blood oranges.

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Satoru Kondo, Kentaro Hiraoka, Shozo Kobayashi, Chikako Honda, and Norihiko Terahara

Cyanidin 3-galactoside was the primary anthocyanin in red `Tsugaru' apples [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.]. The concentration of cyanidin 3-galactoside in the skin decreased from 20 to 62 days after full bloom (DAFB), then increased rapidly after 104 DAFB. Small amounts of cyanidin 3-arabinoside and cyanidin 3-glucoside were detected at 122 and 133 DAFB (harvest). The expression of five anthocyanin biosynthetic genes of chalcone synthase (MdCHS), flavanone 3-hydroxylase (MdF3H), dihydroflavonol 4-reductase (pDFR), anthocyanidin synthase (MdANS), and UDP glucose-flavonoid 3-O-glucosyltransferase (pUFGluT) was examined in the skin of red and nonred apples. In general, the expression of anthocyanin biosynthetic genes in red apples was strong in juvenile and ripening stages. The expression of MdCHS, MdF3H, pDFR, and MdANS was observed before ripening stage when anthocyanin was not detected. In contrast, the expression of pUFGluT was detected in the development stage only when anthocyanin was detected. However, the expression of all five genes was observed at 20 DAFB in fruit bagged after fertilization, and anthocyanin was not detected. The expression of MdCHS, MdF3H, pDFR, and MdANS, excluding pUFGluT, was detected at 98 DAFB in fruit bagged after 30 DAFB, and anthocyanin was not detected. These results suggest that pUFGluT may be closely related to the anthocyanin expression in apple skin at the ripening stage.

Open access

Yingtao Ma, Mengmeng Zhao, Hongxia Wu, Congying Yuan, Huiyun Li, and Yanzhao Zhang

), phenylalanine ammonialyase (PAL), dihydroflavonol 4-reductase (DFR), chalcone isomerase (CHI), anthocyanidin synthase (ANS), flavonoid 3-hydroxylase (F3H), and UDP-glucose: flavonoid-3-O-glucosyltransferase (UFGT) ( Jaakola, 2013 ; Kobayashi et al., 2001

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John R. Stommel and Judith M. Dumm

to a colorless leucoanthocyanidin by dihydroflavonol 4-reductase (DFR), the first committed enzyme of anthocyanin biosynthesis. Leucoanthocyanidins are converted to colored anthocyanidins by anthocyanidin synthase (ANS). UDP-glucose-flavonoid 3-O-glucosyltransferase

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Yuji Yamada, Masayoshi Nakayama, Hiromitsu Shibata, Sanae Kishimoto, and Takashi Ikeda

705847)], and UDP glucose: flavonoid 3-O-glucosyltransferase [ UFGT ; (FJ705848)]; and the transcription factor genes MYB (AJ608992), bHLH (no accession number), and WDR (no accession number). Specific primer sets for PAL , F3′5′H , ANS , MYB

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Ben-Hong Wu, Ning Niu, Ji-Hu Li, and Shao-Hua Li

stress ( Deytieux et al., 2007 ; Grimplet et al., 2009 ; Martínez-Esteso et al., 2011 ). We have recently published our findings that in addition to proteins involved in secondary metabolism [e.g., UDP-glucose flavonoid 3- O -glucosyltransferase (UFGT

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Renata Koyama, Adriane Marinho de Assis, Lilian Yukari Yamamoto, Wellington Fernando Borges, Rogério de Sá Borges, Sandra Helena Prudêncio, and Sergio Ruffo Roberto

with it having a role in the control of berry ripening ( Wheeler et al., 2009 ), resulting from increased expression of the key anthocyanin pathway enzyme UDP-glucose:flavonoid 3-O-glucosyltransferase ( He et al., 2010 ). Thus, the application of

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Hongmei Ma, Margaret Pooler, and Robert Griesbach

biosynthesis genes. Six enzymes are generally involved in the anthocyanin biosynthesis pathway: chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), and UDP-glucose-flavonoid