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RUS6,一种含有 DUF647 的蛋白,在拟南芥的早期胚胎发育中是必需的。

RUS6, a DUF647-containing protein, is essential for early embryonic development in Arabidopsis thaliana.

机构信息

Department of Biology, San Francisco State University, CA, 94132, San Francisco, USA.

出版信息

BMC Plant Biol. 2021 May 25;21(1):232. doi: 10.1186/s12870-021-03011-8.

Abstract

BACKGROUND

The Arabidopsis RUS (ROOT UV-B SENSITIVE) gene family contains six members, each of which encodes a protein containing a DUF647 (domain of unknown function 647) that is commonly found in eukaryotes. Previous studies have demonstrated that RUS1 and RUS2 play critical roles in early seedling development. All six RUS genes are expressed throughout the plant, but little is known about the functional roles of RUS3, RUS4, RUS5 and RUS6.

RESULTS

We used a reverse-genetic approach to identify knockout mutants for RUS3, RUS4, RUS5 and RUS6. Each mutant was confirmed by direct DNA sequencing and genetic segregation analysis. No visible phenotypic differences were observed in rus3, rus4, or rus5 knockout mutants under standard growth conditions, but rus6 knockout mutants displayed a strong embryo-lethal phenotype. Two independent knockout lines for RUS6 were characterized. The rus6 mutations could only be maintained through a heterozygote, because rus6 homozygous mutants did not survive. Closer examinations of homozygous rus6 embryos from rus6/ + parent plants revealed that RUS6 is required for early embryo development. Loss of RUS6 resulted in embryo lethality, specifically at the mid-globular stage. The embryo-lethality phenotype was complemented by a RUS6::RUS6-GFP transgene, and GFP signal was detected throughout the embryo. Histological analyses with the β-glucuronidase reporter gene driven by the RUS6 promoter showed tissue- and development-specific expression of RUS6, which was highest in floral tissues.

CONCLUSION

Our data revealed that RUS6 is essential for early embryo development in Arabidopsis, and that the RUS gene family functions in multiple stages of plant development.

摘要

背景

拟南芥 RUS(ROOT UV-B SENSITIVE)基因家族包含六个成员,每个成员都编码一种含有 DUF647(未知功能域 647)的蛋白质,该蛋白质在真核生物中普遍存在。先前的研究表明,RUS1 和 RUS2 在早期幼苗发育中发挥关键作用。六个 RUS 基因在整个植物中都有表达,但对 RUS3、RUS4、RUS5 和 RUS6 的功能作用知之甚少。

结果

我们使用反向遗传学方法鉴定了 RUS3、RUS4、RUS5 和 RUS6 的敲除突变体。通过直接 DNA 测序和遗传分离分析证实了每个突变体。在标准生长条件下,rus3、rus4 或 rus5 敲除突变体没有观察到明显的表型差异,但 rus6 敲除突变体表现出强烈的胚胎致死表型。对两个独立的 RUS6 敲除系进行了表征。rus6 突变只能通过杂合子维持,因为 rus6 纯合突变体不能存活。对来自 rus6/ + 亲本植物的纯合 rus6 胚胎进行更仔细的检查表明,RUS6 是早期胚胎发育所必需的。RUS6 的缺失导致胚胎致死,特别是在中球阶段。RUS6::RUS6-GFP 转基因可互补 rus6 敲除突变体的胚胎致死表型,并且在整个胚胎中都检测到 GFP 信号。用 RUS6 启动子驱动的 β-葡萄糖醛酸酶报告基因进行的组织学分析显示,RUS6 的表达具有组织和发育特异性,在花组织中表达最高。

结论

我们的数据表明,RUS6 对拟南芥早期胚胎发育是必不可少的,并且 RUS 基因家族在植物发育的多个阶段发挥作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/324e/8146622/69aea07cc7e2/12870_2021_3011_Fig1_HTML.jpg

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