Identification of differentially expressed genes in the flesh of blood and common oranges
Identification of differentially expressed genes in the flesh of blood and common oranges
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DOI：
10.1007/s11295-007-0111-3
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发表时间：
2008-04-01
影响因子：
2.4
通讯作者：
Recupero, Reforgiato Giuseppe
中科院分区：
文献类型：
--
作者：
Licciardello, Concetta;Russo, Maria Patrizia;Recupero, Reforgiato Giuseppe
本研究的目的是鉴定血橙与普通橙相比差异表达的基因，并鉴定血橙果肉中上调的花青素途径基因。使用来自摩罗（一种血橙橙子）的珠心系（58-8D-1）的果肉的RNA作为测试物和来自Cadenera（一种普通橙子）的RNA作为驱动物，构建了1，248个克隆的消减互补DNA文库。经反向北方筛选，共发现230个克隆在血橙子中表达上调，30个克隆在普通金黄色中表达上调。序列分析鉴定了参与花青素苷途径的基因，包括编码生物合成酶如苯丙氨酸解氨酶、查耳酮合酶、二氢黄酮醇-4-还原酶、花青素合酶、UDP：葡萄糖类黄酮3-O-葡糖基转移酶、谷胱甘肽S-转移酶和编码碱性螺旋环受阻蛋白的调节基因，而其他基因与初级代谢、风味生物合成、信号转导机制，和防守一些序列被归类为未知和未命名的，而另一些则未分类。半定量逆转录-聚合酶链反应（PCR）和定量实时PCR被用来确认不同功能类别的候选基因的差异表达模式。一些基因的表达和血橙成熟过程之间的相关性进行了鉴定。
The objective of this research was the identification of genes differentially expressed in blood oranges compared to common oranges and the identification of anthocyanin pathway genes that are up-regulated in flesh of blood oranges. A subtracted complementary DNA library of 1,248 clones was constructed using RNA from the flesh of a nucellar line (58-8D-1) of Moro (a blood orange) as tester and from Cadenera (a common orange) as driver. After screening by reverse Northern, a total of 230 clones were found to be up-regulated in blood orange, while 30 were up-regulated in the common blond one. Sequence analysis identified genes involved in the anthocyanin pathway including genes encoding biosynthetic enzymes like phenylalanine ammonialyase, chalcone synthase, dihydroflavonol-4-reductase, anthocyanidin synthase, UDP: glucose flavonoid 3-O-glucosyltransferase, glutathione S-transferase, and a regulatory gene encoding a basic Helix-Loop-Helix protein, while others were related to primary metabolism, flavor biosynthesis, signal transduction mechanisms, and defense. Some sequences were classified as unknown and unnamed and some others were unclassified. Semiquantitative reverse transcription-polymerase chain reaction (PCR) and quantitative real-time PCR were used to confirm the differential expression patterns of selected candidate genes of different functional classes. Correlations between the expression of some genes and the processes involved in the ripening of blood oranges were identified.