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.
Paola S. Cotroneo, Maria P. Russo, Manuela Ciuni, Giuseppe Reforgiato Recupero, and Angela R. Lo Piero
Dineshkumar Selvaraj, Sherif Sherif, Mohd Sabri Pak Dek, Gopinadhan Paliyath, Islam El-Sharkawy, and Jayasankar Subramanian
ABGs, designated Pd-PAL , Pd-CHS , Pd-DFR , Pd-ANS , and Pd-UFGT (GenBank accession numbers are listed in Supplemental Table 2 ) were isolated from frozen fruit skin tissues (at the commercial harvesting date) using the reverse transcription PCR
W.J. Steyn, D.M. Holcroft, S.J.E. Wand, and G. Jacobs
Changes in activity of phenylalanine ammonia-lyase (PAL) and UDPGalactose: flavonoid-3-o-glycosyltransferase (UFGT) during the development of pear (Pyrus communis L.) fruit and in response to cold fronts were assessed and related to changes in red color. Red and blushed pear cultivars attained maximum redness and highest anthocyanin concentrations in immature fruit. Red color generally faded toward harvest. UFGT activity increased over fruit development and was apparently not limiting to color development. However, the fading of red color and the decreasing level of phenolic compounds toward harvest might relate to decreasing PAL activity. Skin color and enzyme activity in the red pear `Bon Rouge' displayed little responsiveness to low temperatures. In contrast, low temperatures increased red color and activity of both PAL and UFGT in the blushed pear `Rosemarie'. Consistent with the general pigmentation pattern described above, the effect of temperature on enzyme activity was much greater early during fruit development than in the week before harvest.
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.
Keizo Yonemori, Junya Yoshida, Ayako Ikegami, Akihiko Sato, Masahiko Yamada, and Akira Kitajima
Pollination-constant and non-astringent (PCNA)-type persimmon has probably originated from astringent (non-PCNA)-type as a mutant that terminates condensed tannin accumulation at an early stage of fruit development. This trait is confirmed to be recessive and is controlled by a single locus. Since PCNA-type fruit stops tannin accumulation at an early stage, comparison of the gene expressions between PCNA- and non-PCNA-type will reveal the genes conferring condensed tannin accumulation in persimmon fruit. We performed suppression subtractive hybridization (SSH) analysis for detecting differentially expressed genes in non-PCNA-type fruit using BC1 offspring from a cross between PCNA `Fuyu' and non-PCNA “275-13” (F1 progeny derived from non-PCNA `Aizumishirazu' × PCNA `Taishu'). Fruits from seven individuals of PCNA or non-PCNA offspring in BC1 were sampled at early two stages of fruit development and total RNA was extracted by hot borate method from each fruit of different stage. Then, RNA was pooled as PCNA or non-PCNA bulk at two stages and cDNA was synthesized from each bulk for SSH analysis. A total of 5000 clones expressed differentially in non-PCNA-type fruit were picked from SSH library of two stages and 198 positive clones confirmed by differential screening were sequenced. The homologous sequences for the genes involved in flavonoid biosynthesis (CHS, CHI, F3H, F3'5'H, DFR, UFGT, and ANS) were obtained from the clones. The genes that are not considered to be involved in flavonoid biosynthesis so far (SCPL and DHQ) were also detected with high frequencies. We will discuss the role of these genes for condensed tannin accumulation in persimmon fruit.
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
David A. Felicetti and Larry E. Schrader
epicatechin concentrations increased. Both anthocyanins and epicatechin are synthesized from cyanidin. UDP-glycosyl:flavonoid-3-O-glycosyltransferases (UFGTs) transfer glycosides to cyanidin to form anthocyanins, whereas anthocyanidin reductase (ANR) reduces
Xiucai Fan, Renzong Zhao, Qianqian Wang, Chonghuai Liu, and Jinggui Fang
genes are involved in the regulation of anthocyanin biosynthesis in ‘Kyoho’ grape via the expression of the UFGT gene. Moreover, the transcription factors VvmybA1 , VvmybA2 , VvmybA3 , VlmybA1-1 , VlmybA1-2 , and VlmybA2 have been shown to
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
Peter M.A. Toivonen, Jared Stoochnoff, Kevin Usher, Changwen Lu, Paul A. Wiersma, and Chunhua Zhou
ultraviolet response and to regulation of genes associated with the anthocyanin pathway (CHS, CHI, F3H, DFR, LDOX, UFGT, MYB1, MYB8, UVR8, bHLH3, bHLH33, COP1, and HY5). Materials and Methods Preharvest treatments in the orchard. A commercial ‘Ambrosia’ apple