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Complex nested promoters control tissue-specific expression of acetyl-CoA carboxylase genes in wheat

机译:复杂的嵌套启动子控制小麦中乙酰辅酶A羧化酶基因的组织特异性表达

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摘要

Cis-acting regulatory elements of the wheat acetyl-CoA carboxylase (ACC) gene family were identified by comparing the promoter activity of 5′ end gene fragments fused to a reporter gene in two transient expression systems: wheat protoplasts and epidermal cells of mature embryos. Expression of the plastid and the cytosolic ACC genes is each driven by two nested promoters responsible for the synthesis of two transcript types. The internal promoter is located in an intron removed from transcripts originating at the first promoter. These complex promoters, which are different for the cytosolic and plastid ACC genes, control tissue-specific expression of the enzymatic activity supplying cytosolic, plastid, and mitochondrial pools of malonyl-CoA. The activity of one such complex promoter, driving expression of one of the cytosolic ACC genes, was studied throughout development of transgenic wheat plants carrying a full-length promoter–reporter gene fusion. High activity of the promoter was detected in the coleoptile, in the upper sheath section of the leaf, on the top surface of the ovary, in some sections of the main veins in the lemma and glume, and in abaxial epidermis hair cells of the lemma, glume, and rachis. The findings are consistent with the developmental and environmental requirements for very-long-chain fatty acids and flavonoids, whose synthesis begins with the ACC reaction in the cytosol of these specific cell types.
机译:小麦乙酰辅酶A羧化酶(ACC)基因家族的顺式调控元件是通过在两个原生表达系统(小麦原生质体和成熟胚的表皮细胞)中比较与报告基因融合的5'端基因片段的启动子活性来鉴定的。质体和胞质ACC基因的表达各自由两个嵌套的启动子驱动,这些启动子负责两种转录物类型的合成。内部启动子位于一个内含子中,该内含子已从源自第一个启动子的转录本中除去。这些复杂的启动子(对于胞质和质体ACC基因而言是不同的),控制提供丙二酰CoA的胞质,质体和线粒体池的酶活性的组织特异性表达。在携带全长启动子-报告基因融合的转基因小麦植株的整个发育过程中,都研究了一种这样的复杂启动子的活性,该启动子驱动了一个胞质ACC基因的表达。在胚芽鞘,叶片的上部鞘部分,子房的顶表面,外膜和颖壳的主要静脉的某些部分以及外膜的背面表皮毛细胞中检测到启动子的高活性。 ,片状和片状。这些发现与超长链脂肪酸和类黄酮的开发和环境要求是一致的,它们的合成始于这些特定细胞类型的细胞质中的ACC反应。

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