CER16 Inhibits Post-Transcriptional Gene Silencing of CER3 to Regulate Alkane Biosynthesis.,Plant Physiology

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CER16 Inhibits Post-Transcriptional Gene Silencing of CER3 to Regulate Alkane Biosynthesis.
Plant Physiology ( IF 6.5 ) Pub Date : 2020-01-15 , DOI: 10.1104/pp.19.01002
Xianpeng Yang ₁ , Tao Feng ₁ , Shipeng Li _{1,

2} , Huayan Zhao ₃ , Shuangshuang Zhao ₄ , Changle Ma ₄ , Matthew A Jenks ₅ , Shiyou Lü ₆

Affiliation

The aerial surfaces of land plants have a protective layer of cuticular wax. Alkanes are common components of these waxes, and their abundance is affected by a range of stresses. The CER16 protein has been implicated in alkane biosynthesis in the cuticular wax of Arabidopsis (Arabidopsis thaliana). Here, we identified two new mutant alleles of CER16 in Arabidopsis resulting in production of less wax with dramatically fewer alkanes than the wild type. Map-based cloning with genetic analysis revealed that the cer16 phenotype was caused by complete loss of AT5G44150, encoding a protein with no known domains or motifs. Comparative transcriptomic analysis revealed that transcripts of CER3, previously shown to play a principal role in alkane production, were markedly reduced in the cer16 mutants. To define the relationship between CER3 and CER16, we transformed the full CER3 gene into a cer16 mutant. Transgenic CER3 expression was silenced, and levels of small interfering RNAs targeting CER3 were significantly increased. Mutating two major components of the RNA-silencing machinery in a cer16 genetic background restored CER3 transcript levels to wild-type levels, with the stems restored to wild-type glaucousness. We suggest that CER16 deficiency induces post-transcriptional gene silencing of both endogenous and exogenous expression of CER3.

中文翻译：

CER16 抑制 CER3 转录后基因沉默以调节烷烃生物合成。

陆地植物的气生表面有一层角质蜡保护层。烷烃是这些蜡的常见成分，其丰度受到一系列应力的影响。 CER16 蛋白参与拟南芥 (Arabidopsis thaliana) 角质层蜡的烷烃生物合成。在这里，我们在拟南芥中发现了两个新的 CER16 突变等位基因，与野生型相比，其产生的蜡量和烷烃数量显着减少。基于图谱的克隆和遗传分析表明，cer16 表型是由 AT5G44150 完全缺失引起的，AT5G44150 编码一种没有已知结构域或基序的蛋白质。比较转录组学分析表明，之前显示在烷烃生产中起主要作用的 CER3 转录本在 cer16 突变体中显着减少。为了定义 CER3 和 CER16 之间的关系，我们将完整的 CER3 基因转化为 cer16 突变体。转基因CER3表达被沉默，靶向CER3的小干扰RNA的水平显着增加。在 cer16 遗传背景中突变 RNA 沉默机制的两个主要组件，可以将 CER3 转录水平恢复到野生型水平，同时茎也恢复到野生型的青色。我们认为 CER16 缺陷会诱导 CER3 内源和外源表达的转录后基因沉默。

更新日期：2020-03-03

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