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揭示拟南芥AT1G29480——一个仅限于十字花科基因组的神秘基因座。

Shedding light on AT1G29480 of -An enigmatic locus restricted to Brassicacean genomes.

作者信息

Billakurthi Kumari, Schulze Stefanie, Schulz Eva Lena Marie, Sage Tammy L, Schreier Tina B, Hibberd Julian M, Ludwig Martha, Westhoff Peter

机构信息

Institute of Plant Molecular and Developmental Biology Universitätsstrasse 1, Heinrich-Heine-University Duesseldorf Germany.

Cluster of Excellence on Plant Sciences 'From Complex Traits Towards Synthetic Modules' Düsseldorf-Cologne Germany.

出版信息

Plant Direct. 2022 Oct 17;6(10):e455. doi: 10.1002/pld3.455. eCollection 2022 Oct.

Abstract

A key feature of C Kranz anatomy is the presence of an enlarged, photosynthetically highly active bundle sheath whose cells contain large numbers of chloroplasts. With the aim to identify novel candidate regulators of C bundle sheath development, we performed an activation tagging screen with . The reporter gene used encoded a chloroplast-targeted GFP protein preferentially expressed in the bundle sheath, and the promoter of the C phosphopyruvate carboxylase gene from served as activation tag because of its activity in all chlorenchymatous tissues of . Primary mutants were selected based on their GFP signal intensity, and one stable mutant named with a significant increase in GFP fluorescence intensity was obtained. Despite the increased GFP signal, showed no alterations to bundle sheath anatomy. The causal locus, AT1G29480, is specific to the Brassicaceae with its second exon being conserved. Overexpression and reconstitution studies confirmed that AT1G29480, and specifically its second exon, were sufficient for the enhanced GFP phenotype, which was not dependent on translation of the locus or its parts into protein. We conclude, therefore, that the AT1G29480 locus enhances the GFP reporter gene activity via an RNA-based mechanism.

摘要

C4 花环解剖结构的一个关键特征是存在一个增大的、光合活性高的维管束鞘,其细胞含有大量叶绿体。为了鉴定 C4 维管束鞘发育的新型候选调节因子,我们用……进行了激活标签筛选。所使用的报告基因编码一种优先在维管束鞘中表达的叶绿体靶向 GFP 蛋白,并且来自……的 C4 磷酸烯醇式丙酮酸羧化酶基因的启动子因其在……所有绿色组织中的活性而用作激活标签。根据 GFP 信号强度选择初级突变体,获得了一个名为……的稳定突变体,其 GFP 荧光强度显著增加。尽管 GFP 信号增强,但……的维管束鞘解剖结构没有改变。因果基因座 AT1G29480 是十字花科特有的,其第二个外显子是保守的。过表达和重组研究证实,AT1G29480,特别是其第二个外显子,足以产生增强的 GFP 表型,这并不依赖于该基因座或其部分翻译成蛋白质。因此,我们得出结论,AT1G29480 基因座通过基于 RNA 的机制增强了 GFP 报告基因的活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/215a/9576117/c9246d983fb1/PLD3-6-e455-g002.jpg

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