Phosphorus (Pi) plays a crucial role in the growth and development of plants. Membrane lipid regulation is one of the main mechanisms underlying plant adaptation to Pi deficiency. Previously, the high tolerance to low-Pi stress was justified in an elite line, MSDZ 109, which was obtained from . To better understand the mechanism underlying high adaptation to low-Pi stress, currently, lipidomic and transcriptomic analysis, as well as CRISPR/Cas9 and -overexpressing methodologies were comprehensively integrated into a strategy for elucidating the high tolerance to low-Pi stress. Totally, 770 differential metabolites were identified from the roots between the low-Pi and stress-free, belonging to 21 sub-classes of lipid compounds. Fatty acids (FA) constituted the predominant lipid component, accounting for approximately 50%–60% of the total lipids, and triglycerides (TAG) ranked the second, comprising around 12% of the total, consecutively followed by phosphatidylcholine (PC) and diacylglycerol (DAG) with approximately 10% and 8% of the total, respectively. The synchronous transcriptomic analysis revealed a significant up-regulation of genes related to glycerophospholipid and glycerolipid metabolism, specifically those (, etc.) involved in phospholipid and galactosyl synthesis in response to low-Pi stress. GUS fusing reporter assay showed that promoter induced GUS gene expression and demonstrated initiation activity. Based on expression analysis, a dual-luciferase reporter assay, as well as yeast one-hybrid (Y1H) identification, was justified to bind with the promoter and positively regulate plant tolerance to low-Pi stress. To further elucidate the role of , CRISPR/Cas9 and -overexpressing vectors were successfully introduced into apple (‘Royal Gala’) calli. Interestingly, the -KO line calli exhibited the remarkable decreases in the contents of phosphodiesterase (PDE), activity, as well as the contents of total Pi, and Pi in comparison with those of the wild type. Conversely, -OE ones demonstrated the significant elevation in Pi accumulations, further justifying its potential role in Pi remobilization in apple. Therefore, substantially involves elevating low-Pi tolerance via promoting Pi release in .

中文翻译：

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脂质组和转录组的关联分析强调了MmGDPD1参与调节Malus mandshurica的低磷耐受性

磷（Pi）在植物的生长和发育中起着至关重要的作用。膜脂调节是植物适应磷缺乏的主要机制之一。此前，对低 Pi 压力的高耐受性在精英品系 MSDZ 109 中得到了证明，该品系来自 .为了更好地理解低Pi胁迫高适应机制，目前，脂质组学和转录组学分析以及CRISPR/Cas9和过表达方法被全面整合到阐明低Pi胁迫高耐受性的策略中。总共从低Pi和无胁迫的根中鉴定出770种差异代谢物，属于21个脂质化合物亚类。脂肪酸（FA）是主要的脂质成分，约占总脂质的50%~60%，甘油三酯（TAG）位居第二，约占总量的12%，其次是磷脂酰胆碱（PC）和二酰甘油(DAG) 分别约占总数的 10% 和 8%。同步转录组分析揭示了与甘油磷脂和甘油脂代谢相关的基因显着上调，特别是那些参与磷脂和半乳糖基合成以响应低Pi应激的基因。 GUS 融合报告基因测定表明启动子诱导 GUS 基因表达并表现出起始活性。基于表达分析，双荧光素酶报告基因测定以及酵母单杂交体 (Y1H) 鉴定被证明与启动子结合并正向调节植物对低 Pi 胁迫的耐受性。为了进一步阐明 的作用，将 CRISPR/Cas9 和 过表达载体成功引入苹果（“Royal Gala”）愈伤组织中。有趣的是，与野生型相比，-KO系愈伤组织表现出磷酸二酯酶(PDE)含量、活性以及总Pi和Pi含量的显着降低。相反，-OE 表现出 Pi 积累的显着升高，进一步证明了其在苹果中 Pi 再动员中的潜在作用。因此，主要涉及通过促进 Pi 释放来提高低 Pi 耐受性。