花生长链酰基辅酶A合成酶6基因(<i>LACS</i>6)的克隆、鉴定与组织表达

徐日荣; 陈湘瑜; 唐兆秀

doi:10.19303/j.issn.1008-0384.2016.11.005

花生长链酰基辅酶A合成酶6基因(LACS6)的克隆、鉴定与组织表达

doi: 10.19303/j.issn.1008-0384.2016.11.005

徐日荣^{1, 2,},
陈湘瑜^{1, 2},
唐兆秀^{1, 2, ,}

1.
福建省农业科学院作物研究所, 福建福州 350013
2.
国家农业部闽台农作物种质资源利用重点开放实验室, 福建福州 350013

基金项目:

福建省科技计划项目——省属公益类科研院所基本科研专项 2014R1026-4

详细信息

作者简介:
徐日荣(1979-),男,硕士,副研究员,研究方向:花生遗传育种(E-mail:rirongxu@163.com)

通讯作者:
唐兆秀(1958-),男,研究员,研究方向:花生遗传育种(E-mail:tzxfz@163.com)

中图分类号: S565.2
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- 被引次数: 0
出版历程
- 收稿日期: 2016-05-12
- 修回日期: 2016-09-19
- 刊出日期: 2016-11-01

Cloning, Verification and Tissue Expression of Long-chain Acyl-CoA Synthetase 6 (LACS6) Gene in Peanut (Arachis hypogaea L.)

XU Ri-rong^{1, 2
,},
CHEN Xiang-yu^{1, 2},
TANG Zhao-xiu^{1, 2
, ,}

1.
Institute of Crop Sciences, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350013, China
2.
Key Laboratory of Crop Germplasm Utilization between Fujian and Taiwan, Ministry of Agriculture, Fuzhou, Fujian 350013, China

摘要

摘要: 长链酰基辅酶A合成酶（long chain acyl-CoA synthetase：LACS）是油脂代谢的重要催化酶。为揭示花生脂肪酸代谢机理，采用RT-PCR技术，首次从花生Arachis hypogaea L.克隆到LACS6（GenBank登录号：KU301860），分析该基因的结构组成，预测编码氨基酸与其他植物的同源性，采用Real-Time PCR技术对LACS6的组织表达进行研究。结果显示，花生LACS6基因全长2 116 bp，包含2 088 bp的ORF，编码695个氨基酸，有23个外显子和22个内含子。氨基酸序列比对显示花生LACS6有真核生物酰基辅酶A合成酶保守结构域，并含有保守的激活位点和绑定位点。同源性分析发现花生LACS6与鹰嘴豆、绿豆、大豆、梅等13种物种的氨基酸一致性在79%~87%，进化树分析显示，花生LACS6与鹰嘴豆等豆科植物亲缘较近。实时荧光PCR分析表明，花生LACS6在花生根、茎、叶、子房柄、仁和花等组织均有表达，且差异明显。子房柄和花的表达量极高，与根、茎、叶和仁等组织有极显著差异，花生LACS6组织的表达量大小排序为花 > 子房柄 > 叶 > 仁 > 茎 > 根。本研究结果为揭示花生脂肪酸代谢和品质改良提供理论依据。
- 花生 /
- 长链酰基辅酶A合成酶 /
- 组织表达 /
- 油脂
Abstract: Long-chain acyl coenzyme A synthetase (LACS) is one of the important catalytic lipid metabolic enzymes in peanut plants. To understand the mechanism of the fatty acid metabolism in peanut (Arachis hypogaea L.), the cDNA of LACS6 gene was cloned by RT-PCR (GenBank accession number:KU301860). Its structure was analyzed, homology with other plants compared, and mRNA expression in different tissues obtained using real-time PCR. The results showed that the cDNA was 2,116 bp including an open reading frame of 2,088 bp, encoded 695 amino acids, and had 23 exons and 22 introns. Its amino acid sequence alignment showed a conserved domain of eukaryotic long-chain fatty acid CoA synthetase, as well as conserved activation and binding sites. The homologies of LACS6 gene with the genes from Cicer arietinum, Vigna radiate, Glycine max, Prunus mume and others were between 79% and 87%. The evolutionary tree indicated that the gene was close to those from leguminous plants, such as chickpeas. The real-time PCR analysis showed that it was differentially expressed in the ovary stalks, roots, stems, leaves, flowers and kernels on a peanut plant. The ovary stalks and flowers had the highest expression among all,and with significant differences,The order of expression ranked as flowers > ovary stalks > leaves > kernels > stems > roots. The information obtained would benefit further studies on the fatty acid metabolism and quality improvement for peanut.
- peanut (Arachis hypogaea L.) /
- long-chain acyl CoA synthetase /
- tissue expression /
- lipid

HTML全文

图 1 花生AhLACS6基因PCR电泳结果

注：1为AhLACS6基因；M为 DL2000 DNA Marker。

Figure 1. Electrophoresis on LACS6 gene of A.hypogaea (Ah)

下载: 全尺寸图片幻灯片

图 2 花生AhLACS6基因的核酸序列及预测氨基酸序列

Figure 2. Nucleic acid and amino acid sequences of AhLACS6 gene

下载: 全尺寸图片幻灯片

图 3 花生AhLACS6基因在基因组中结构

Figure 3. Genome structure of AhLACS6 gene

下载: 全尺寸图片幻灯片

图 4 AhLACS6与其他植物LACS6的多重比较

注：下划线为酰基激活酶AAE保守序列。

Figure 4. Multiple comparisons on AhLACS6 gene and other plant LACS6 genes

下载: 全尺寸图片幻灯片

图 5 AhLACS6与其他植物进化树分析

Figure 5. Evolutionary tree analysis on AhLACS6 gene and other plant LACS6 genes

下载: 全尺寸图片幻灯片

图 6 AhLACS6在不同组织的荧光定量PCR分析

注：不同大写字母表示差异极显著(P＜0.01)。

Figure 6. Fluorescence quantitative PCR analysis on AhLACS6 gene in various plant organs

下载: 全尺寸图片幻灯片

表 1 AhLACS6与其他植物LACS6的一致性

Table 1. Homology of AhLACS6 gene and other plant LACS6 genes

	物种	登录号	一致性 /%
鹰嘴豆	Cicer arietinum	XP_004496750.1	87
绿豆	Vigna radiata	XP_014524185.1	86
大豆	Glycine max	XP_003555336.1	86
梅	Prunus mume	XP_008222945.1	85
葡萄	Vitis vinifera	XP_002277936.1	84
蓖麻	Ricinus communis	NP_001310618.1	84
白梨	Pyrus x bretschneideri	XP_009364219.1	85
苹果	Malus domestica	XP_008390867.1	85
枣	Ziziphus jujuba	XP_015888035.1	84
野草莓	Fragaria vesca	XP_004296905.1	83
杨	Populus trichocarpa	XP_002315784.2	82
黄瓜	Cucumis sativus	XP_004134142.1	81
欧洲油菜	Brassica napus	XP_013735920.1	79

下载: 导出CSV

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