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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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Mingjun Li, Xuesen Chen, Pingping Wang, and Fengwang Ma

new insight into that process in apple, we systematically investigated here the accumulation of AsA, mRNA expression of genes involved in its biosynthesis as well as recycling, and the relationships among gene expression, enzyme activity, and AsA

Free access

Xi Shan, Heng Zhou, Ting Sang, Sheng Shu, Jin Sun, and Shirong Guo

We investigated the effects of exogenous spermidine (Spd) on the carbohydrate, nitrogen (N), and endogenous polyamine status of tomato (Solanum lycopersicum) seedlings exposed to high-temperature stress [38/28 °C (day/night)]. High-temperature stress reduced the contents of pyruvate and succinate and inhibited plant growth. The application of exogenous Spd alleviated the inhibition of plant growth induced by high temperature, and also led to an increase in pyruvate, citrate, and succinate levels. High temperature markedly increased the NH4 +-N content and reduced the activities of nitrate reductase (NR), glutamine synthetase (GS), and glutamate dehydrogenase (GDH). Spd significantly alleviated the negative effects on NH4 +-N assimilation induced by high-temperature stress. Moreover, Spd significantly increased the activities of NR and GDH in the high-temperature-stressed tomato leaves. In contrast, Spd application to high-temperature-stressed plant leaves counteracted high-temperature-induced mRNA expression changes in N metabolism. Spd significantly upregulated the transcriptional levels of NR, nitrite reductase, GS, GDH, and glutamate synthase (GOGAT). In addition, exogenous Spd significantly increased endogenous polyamines. These results suggest that Spd could improve carbohydrate and N status through regulating the gene expression and activity of key enzymes for N metabolism, thus confers the tolerance to high temperature on tomato seedlings.

Free access

Denise M. Tieman and Avtar K. Handa

The growth of tomato fruit is the result of cell division early in development followed by cell expansion until the onset of ripening. We have utilized the mRNA differential display technique to clone genes differentially expressed in 10- and 20-day-old tomato fruit, when most fruit cells are undergoing a transition to growth by cell expansion. Of 1753 total bands observed using 30 independent primer sets, 31 differential display bands were obtained only in either 10-or 20-day - old fruit RNAs. Seven differentially expressed bands from 10-day-old fruit RNAs and six from 20-day-old fruit RNAs were cloned and characterized by sequence analysis and mRNA expression patterns in developing fruit, leaf and root tissues. Two clones had sequence similarities to 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase or threonine deaminase genes, while the remaining clones did not correspond to previously characterized genes. Steady state levels of mRNAs corresponding to seven clones were upregulated between 10 and 20 days of fruit development, while two clones were downregulated during growth and ripening. Most clones also hybridize to mRNA species present in leaf and root tissues. Collectively, these results suggest a transition in gene expression between 10- and 20-day-old fruit development.

Free access

Zhencai Wu and Paul A. Wiersma

Expansins are a class of proteins that stimulate the extension of plant cell walls. Expansins have been found in nearly all growing plant tissues, such as hycopotyls, young seedlings, fibers, internodes, flower petals, and ripening fruits. We isolated two full-length expansin cDNA clones, Pruav-Exp1 and Pruav-Exp2, from sweet cherry (Prunus avium L.) fruit. Pruav-Exp1 has 1048 nucleotides encoding 254 amino acids, while Pruav-Exp2 has 1339 nucleotides encoding 250 amino acids. Deduced amino acid sequences of sweet cherry Pruav-Exp1 and Pruav-Exp2 share 72% identity. A Blast search of the GenBank database with the deduced amino acid sequences of Pruav-Exp1 and Pruav-Exp2 indicated a high sequence identity with other plant expansin genes. Interestingly, Pruav-Exp1 shares 99% identity of amino acid sequence with that of apricot expansin Pav-Exp1. Fragments from the 3' ends of Pruav-Exp1 and Pruav-Exp2 were cloned to generate gene-specific probes. These probes were used to study expansin gene expression in different tissues and during fruit development. Northern blot analysis showed different mRNA expression patterns for each gene. The mRNA of Pruav-Exp1 was expressed at the pink and ripe stages, but not at the early green and yellow stages of fruit development. The mRNA of Pruav-Exp2 was present earlier, from a low level in yellow expanding fruit, increasing to a high level at the pink stage and remaining at this level through the ripe stage. Both mRNAs were also expressed at a low level in flower, but not present in other tissues such as roots, leaves and peduncles. Our study indicates an expansin gene family is present in sweet cherry and suggests that two expansin genes may have different roles during fruit development and ripening.

Free access

Akira Tateishi, Kenji Nagashima, Francis M. Mathooko, Mercy W. Mwaniki, Yasutaka Kubo, Akitsugu Inaba, Shohei Yamaki, and Hiroaki Inoue

Galactosidases are thought to play a key role in cell wall metabolism during fruit growth and ripening. In this study we cloned seven β-galactosidase (β-Gal) cDNAs from japanese pear fruit and designated them PpGAL2, PpGAL3, Pp-GAL4, PpGAL5, PpGAL6, PpGAL7, and PpGAL8, in addition to the previously described JP-GAL hereinafter termed PpGAL1. mRNA expression patterns of these clones were characterized throughout fruit growth and on-tree ripening, and in leaves and shoots in three japanese pear cultivars, `Housui', `Kousui', and `Niitaka'. The shared amino acid sequence identity among the eight japanese pear β-Gal (PpGAL) clones ranged from 50% to 60%. They all contained the putative active site containing consensus sequence pattern G-G-P-[LIVM](2)-x(2)-Q-X-E-N-E-[FY] belonging to glycoside hydrolase family 35. Expression of all the clones was both development- and tissue-specific. PpGAL1 and Pp-GAL4 were only expressed in the ripe fruit while PpGAL2 and PpGAL3 were expressed in both expanding and ripening fruit with their abundance being highest in the ripe fruit. The abundance of PpGAL5, PpGAL6, and PpGAL7 mRNAs was highest in expanding fruit but decreased drastically upon the onset of ripening. PpGAL8 was only detected in very young fruit (15 days after full bloom) and not in expanding and ripening fruit. These results indicate that in japanese pear fruit β-Gal is encoded by a multigene family whose members show distinct and overlapping expression during the various phases of fruit development. Some of the members are not only fruit-specific but also ripening-specific and, therefore, may play a crucial role in cell wall disassembly during japanese pear fruit softening.

Open access

En-chao Liu, Li-fang Niu, Yang Yi, Li-mei Wang, You-wei Ai, Yun Zhao, Hong-xun Wang, and Ting Min

; however, the expression of NnPAL1 was continuously inhibited by MAP, and the relative abundance of the gene was reduced by more than 10 times around day 28. Messenger RNA (mRNA) expression of NnPAL2 in the MAP group increased gradually and peaked at

Open access

Joseph Krystel, Huawei Liu, John Hartung, and Ed Stover

-DNA) insertion was confirmed by PCR and scFv mRNA expression levels were quantified by reverse transcriptase qPCR using the sequence specific primers (primers 1314-F, 1314-R, 1419-F, and 1419-R) listed in Table 1 . For this initial series of phenotypic studies

Free access

Cristián Vela-Hinojosa, Héctor B. Escalona-Buendía, José A. Mendoza-Espinoza, Juan M. Villa-Hernández, Ricardo Lobato-Ortíz, Juan E. Rodríguez-Pérez, and Laura J. Pérez-Flores

) for 20 s, and an extension temperature of 72 °C for 30 s. Thirty cycles were required for all genes, except for LCY-B, which required 32 cycles. Relative mRNA expression was calculated by the 2 -ΔΔCt method ( Villa-Hernández et al., 2013 ). A value of

Free access

Tao Wang, Ruijie Hao, Huitang Pan, Tangren Cheng, and Qixiang Zhang

detect and verify changes in the mRNA expression levels of genes at different developmental stages ( Koo et al., 2010 ; Vaucheret et al., 2004 ) and under various abiotic stress ( Borges et al., 2012 ; Du et al., 2013 ). Normalized quantification of