WHIRLY1 functions in the nucleus to regulate barley leaf development and associated metabolite profiles.
WHIRLY1 functions in the nucleus to regulate barley leaf development and associated metabolite profiles.
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WHIRLY1在细胞核中发挥作用，调节大麦叶片发育和相关代谢产物谱。
DOI：
10.1042/bcj20210810
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发表时间：
2022-03-18
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WHIRLY (WHY) DNA/RNA结合蛋白在叶片发育过程中具有多种功能，但特征不明确。在这里，我们发现WHY1转录本水平在7天龄大麦叶片的碱基中最高。免疫金标记显示，细胞核中WHY1蛋白比叶基部的原质体更丰富。为了鉴定与叶片发育相关的转录本，我们沿着野生型叶片的发育梯度对差异丰富的转录本进行了分层聚类。同样，代谢物谱被用来鉴定代谢物表现出发育梯度。通过对W1-1和W1-7缺失WHY1的大麦品系（W1-1和W1-7）的转录本和代谢物进行比较分析，发现W1-1和W1-7叶片发育的转录本谱基本没有变化。然而，编码与叶绿体发育相关的转录因子的几个转录本的水平存在差异。其中包括调节气孔发育、绿化和叶绿体发育的拟南芥GATA转录因子的大麦同源物NAC1；两个与拟南芥GLK1相似的转录本和两个编码在叶片形态发生和发育中起作用的ARF转录因子的转录本。W1-1和W1-7叶片的叶绿体蛋白含量低于野生型。WHY1基因敲除叶片中三羧酸循环代谢物和GABA含量显著低于野生型。本研究提供的证据表明，WHY1定位于叶碱基的细胞核中，参与调控叶绿体发育的核编码转录物。
The WHIRLY (WHY) DNA/RNA binding proteins fulfil multiple but poorly characterised functions in leaf development. Here, we show that WHY1 transcript levels were highest in the bases of 7-day old barley leaves. Immunogold labelling revealed that the WHY1 protein was more abundant in the nuclei than the proplastids of the leaf bases. To identify transcripts associated with leaf development we conducted hierarchical clustering of differentially abundant transcripts along the developmental gradient of wild-type leaves. Similarly, metabolite profiling was employed to identify metabolites exhibiting a developmental gradient. A comparative analysis of transcripts and metabolites in barley lines (W1–1 and W1–7) lacking WHY1, which show delayed greening compared with the wild type revealed that the transcript profile of leaf development was largely unchanged in W1–1 and W1–7 leaves. However, there were differences in levels of several transcripts encoding transcription factors associated with chloroplast development. These include a barley homologue of the Arabidopsis GATA transcription factor that regulates stomatal development, greening and chloroplast development, NAC1; two transcripts with similarity to Arabidopsis GLK1 and two transcripts encoding ARF transcriptions factors with functions in leaf morphogenesis and development. Chloroplast proteins were less abundant in the W1–1 and W1–7 leaves than the wild type. The levels of tricarboxylic acid cycle metabolites and GABA were significantly lower in WHY1 knockdown leaves than the wild type. This study provides evidence that WHY1 is localised in the nuclei of leaf bases, contributing the regulation of nuclear-encoded transcripts that regulate chloroplast development.