The first intron and promoter of Arabidopsis DIACYLGLYCEROL ACYLTRANSFERASE 1 exert synergistic effects on pollen and embryo lipid accumulation.

IF 9.4 1区 生物学 Q1 Agricultural and Biological Sciences
New Phytologist Pub Date : 2024-11-05 DOI:10.1111/nph.20244
Sean T McGuire, Jay Shockey, Philip D Bates
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引用次数: 0

Abstract

Accumulation of triacylglycerols (TAGs) is crucial during various stages of plant development. In Arabidopsis, two enzymes share overlapping functions to produce TAGs, namely acyl-CoA:diacylglycerol acyltransferase 1 (DGAT1) and phospholipid:diacylglycerol acyltransferase 1 (PDAT1). Loss of function of both genes in a dgat1-1/pdat1-2 double mutant is gametophyte lethal. However, the key regulatory elements controlling tissue-specific expression of either gene has not yet been identified. We transformed a dgat1-1/dgat1-1//PDAT1/pdat1-2 parent with transgenic constructs containing the Arabidopsis DGAT1 promoter fused to the AtDGAT1 open reading frame either with or without the first intron. Triple homozygous plants were obtained, however, in the absence of the DGAT1 first intron anthers fail to fill with pollen, seed yield is c. 10% of wild-type, seed oil content remains reduced (similar to dgat1-1/dgat1-1), and non-Mendelian segregation of the PDAT1/pdat1-2 locus occurs. Whereas plants expressing the AtDGAT1pro:AtDGAT1 transgene containing the first intron mostly recover phenotypes to wild-type. This study establishes that a combination of the promoter and first intron of AtDGAT1 provides the proper context for temporal and tissue-specific expression of AtDGAT1 in pollen. Furthermore, we discuss possible mechanisms of intron mediated regulation and how regulatory elements can be used as genetic tools to functionally replace TAG biosynthetic enzymes in Arabidopsis.

拟南芥二乙酰甘油 ACYLTRANSFERASE 1 的第一个内含子和启动子对花粉和胚胎脂质积累有协同作用。
三酰甘油(TAG)的积累在植物发育的各个阶段都至关重要。在拟南芥中,有两种酶具有产生 TAGs 的重叠功能,即酰基-CoA:二酰甘油酰基转移酶 1(DGAT1)和磷脂:二酰甘油酰基转移酶 1(PDAT1)。dgat1-1/pdat1-2 双突变体中这两个基因的功能缺失会导致配子体致死。然而,控制这两个基因组织特异性表达的关键调控元件尚未确定。我们用含有拟南芥 DGAT1 启动子的转基因构建体转化了 dgat1-1/dgat1-1//PDAT1/pdat1-2 亲本,该构建体融合了带有或不带有第一个内含子的 AtDGAT1 开放阅读框。然而,在没有 DGAT1 第一个内含子的情况下,花药不能充满花粉,种子产量约为野生型的 10%,种子含油量仍然降低(与 dgat1-1/dgat1-1 相似),并且 PDAT1/pdat1-2 基因座出现非孟德尔分离。而表达含有第一个内含子的 AtDGAT1pro:AtDGAT1 转基因的植株,其表型大多恢复为野生型。这项研究证实,AtDGAT1启动子和第一个内含子的结合为AtDGAT1在花粉中的时间和组织特异性表达提供了适当的背景。此外,我们还讨论了内含子介导调控的可能机制,以及如何利用调控元件作为遗传工具,在拟南芥中实现 TAG 生物合成酶的功能替代。
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来源期刊
New Phytologist
New Phytologist PLANT SCIENCES-
CiteScore
17.60
自引率
5.30%
发文量
728
审稿时长
1 months
期刊介绍: New Phytologist is a leading publication that showcases exceptional and groundbreaking research in plant science and its practical applications. With a focus on five distinct sections - Physiology & Development, Environment, Interaction, Evolution, and Transformative Plant Biotechnology - the journal covers a wide array of topics ranging from cellular processes to the impact of global environmental changes. We encourage the use of interdisciplinary approaches, and our content is structured to reflect this. Our journal acknowledges the diverse techniques employed in plant science, including molecular and cell biology, functional genomics, modeling, and system-based approaches, across various subfields.
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