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RNA-seq 数据揭示了参与沙棘浆果果肉中棕榈油酸和油高积累的多基因的协调调控机制。

RNA-seq data reveals a coordinated regulation mechanism of multigenes involved in the high accumulation of palmitoleic acid and oil in sea buckthorn berry pulp.

机构信息

Key Laboratory of Biotechnology and Bioresources Utilization, Ministry of Education, Institute of Plant Resources, Dalian Minzu University, 18 Liaohe West Road, Dalian, 116600, Liaoning, China.

Institute of Berries, Heilongjiang Academy of Agricultural Sciences, 5 Fansheng Street, Suiling, Heilongjiang, 152230, China.

出版信息

BMC Plant Biol. 2019 May 20;19(1):207. doi: 10.1186/s12870-019-1815-x.

Abstract

BACKGROUND

Sea buckthorn is a woody oil crop in which palmitoleic acid (C16:1n7, an omega-7 fatty acid (FA)) contributes approximately 40% of the total FA content in berry pulp (non-seed tissue). However, the molecular mechanisms contributing to the high accumulation of C16:1n7 in developing sea buckthorn berry pulp (SBP) remain poorly understood.

RESULTS

We identified 1737 unigenes associated with lipid metabolism through RNA-sequencing analysis of the four developmental stages of berry pulp in two sea buckthorn lines, 'Za56' and 'TF2-36'; 139 differentially expressed genes were detected between the different berry pulp developmental stages in the two lines. Analyses of the FA composition showed that the C16:1n7 contents were significantly higher in line 'Za56' than in line 'TF2-36' in the mid-late developmental stages of SBP. Additionally, qRT-PCR analyses of 15 genes involved in FA and triacylglycerol (TAG) biosynthesis in both lines revealed that delta9-ACP-desaturase (ACP-Δ9D) competed with 3-ketoacyl-ACP-synthase II (KASII) for the substrate C16:0-ACP and that ACP-Δ9D and delta9-CoA-desaturase (CoA-Δ9D) gene expression positively correlated with C16:1n7 content; KASII and fatty acid elongation 1 (FAE1) gene expression positively correlated with C18:0 content in developing SBP. Specifically, the abundance of ACP-Δ9D and CoA-Δ9D transcripts in line 'Za56', which had a higher C16:1n7 content than line 'TF2-36', suggests that these two genes play an important role in C16:1n7 biosynthesis. Furthermore, the high expressions of the glycerol-3-phosphate dehydrogenase (GPD1) gene and the WRINKLED1 (WRI1) transcription factor contributed to increased biosynthesis of TAG precursor and FAs, respectively, in the early developmental stages of SBP, and the high expression of the diacylglycerol O-acyltransferase 1 (DGAT1) gene increased TAG assembly in the later developmental stages of SBP. Overall, we concluded that increased ACP-Δ9D and CoA-Δ9D levels coupled with decreased KASII and FAE1 activity is a critical event for high C16:1n7 accumulation and that the coordinated high expression of WRI1, GPD1, and DGAT1 genes resulted in high oil accumulation in SBP.

CONCLUSION

Our results provide a scientific basis for understanding the mechanism of high C16:1n7 accumulation in berry pulp (non-seed tissue) and are valuable to the genetic breeding programme for achieving a high quality and yield of SBP oil.

摘要

背景

沙棘是一种木本油料作物,其中棕榈油酸(C16:1n7,一种ω-7 脂肪酸(FA))约占浆果果肉(非种子组织)中总 FA 含量的 40%。然而,高浓度 C16:1n7 在沙棘浆果果肉(SBP)中的积累的分子机制仍知之甚少。

结果

我们通过对两个沙棘品种“Za56”和“TF2-36”的四个发育阶段的浆果果肉进行 RNA-seq 分析,鉴定了 1737 个与脂质代谢相关的基因;在这两个品种中,不同浆果果肉发育阶段之间检测到 139 个差异表达基因。对 FA 组成的分析表明,在 SBP 的中晚期发育阶段,“Za56”线中的 C16:1n7 含量明显高于“TF2-36”线。此外,对这两个品种中参与 FA 和三酰基甘油(TAG)生物合成的 15 个基因的 qRT-PCR 分析表明,Δ9-ACP-脱饱和酶(ACP-Δ9D)与 3-酮酰基-ACP 合酶 II(KASII)竞争 C16:0-ACP 底物,并且 ACP-Δ9D 和 Δ9-CoA-脱饱和酶(CoA-Δ9D)基因表达与 C16:1n7 含量呈正相关;KASII 和脂肪酸延长酶 1(FAE1)基因表达与发育中的 SBP 中 C18:0 含量呈正相关。具体来说,在 C16:1n7 含量高于“TF2-36”线的“Za56”线中,ACP-Δ9D 和 CoA-Δ9D 转录本的丰度表明这两个基因在 C16:1n7 生物合成中发挥重要作用。此外,甘油-3-磷酸脱氢酶(GPD1)基因和 WRINKLED1(WRI1)转录因子的高表达分别有助于 SBP 早期发育阶段 TAG 前体和 FAs 的生物合成,而二酰基甘油 O-酰基转移酶 1(DGAT1)基因的高表达则增加了 SBP 后期发育阶段的 TAG 组装。总的来说,我们得出的结论是,ACP-Δ9D 和 CoA-Δ9D 水平的增加伴随着 KASII 和 FAE1 活性的降低是 C16:1n7 积累增加的关键事件,而 WRI1、GPD1 和 DGAT1 基因的协调高表达导致 SBP 中油的积累增加。

结论

我们的研究结果为理解浆果果肉(非种子组织)中 C16:1n7 积累的机制提供了科学依据,对实现沙棘籽油优质高产的遗传育种计划具有重要价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/18b9/6528223/29c5ea106e5c/12870_2019_1815_Fig1_HTML.jpg

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