Cloning, Characterization and Expression Analysis of a Stearoyl-ACP Desaturase Gene from Arachis Hypogaea

Shanlin Yu, Mingna Chen, Qingli Yang, Hongsheng Zhang
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引用次数: 1

Abstract

SHANLIN YU b, , MINGNA CHEN , QINGLI YANG, HONGSHENG ZHANG a Shandong Peanut Research Institute, Qingdao 266100, China b State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China Abstract Stearoyl-ACP desaturase (SAD), which catalyzes the first desaturation reaction of the most common pathway of unsaturated fatty acid synthesis in plants, plays a key role in determining the ratio of saturated to unsaturated fatty acids. Using Rapid Amplification of cDNA Ends (RACE) and a peanut cDNA library we had previously constructed, we isolated a 1499 bp cDNA of the SAD gene containing a 1218 bp complete open reading fragment (ORF). Sequence analysis revealed that peanut SAD had a high level of nucleotide and amino acid sequence homology to SAD from other plants. Characteristics of the deduced protein were predicted and analyzed using bioinformatic methods. Phylogenetic analysis showed that three different types of delta-9 fatty acid desaturases form separate clades, indicating that the three isozymes may have evolved independently. Expression analysis via real-time PCR indicated that expression levels of the SAD gene were markedly distinct in different peanut tissues and varieties. The results of the expression analysis in this study, combined with existing research, suggest that SAD may be involved in the regulation of plant seed growth and development.
花生硬脂酰acp去饱和酶基因的克隆、表征及表达分析
b南京农业大学作物遗传与种质增效国家重点实验室,南京210095摘要硬脂酰acp去饱和酶(SAD)是催化植物中最常见的不饱和脂肪酸合成途径的第一次去饱和反应,在决定饱和脂肪酸与不饱和脂肪酸的比例中起着关键作用。利用fast Amplification of cDNA Ends (RACE)和我们之前构建的花生cDNA文库,我们分离出了一个1499 bp的SAD基因cDNA,其中包含一个1218 bp的完整开放阅读片段(ORF)。序列分析表明,花生SAD与其他植物的SAD具有较高的核苷酸和氨基酸同源性。利用生物信息学方法预测并分析了所得蛋白的特征。系统发育分析表明,三种不同类型的δ -9脂肪酸去饱和酶形成了不同的分支,表明这三种同工酶可能是独立进化的。实时荧光定量PCR分析表明,SAD基因在不同花生组织和品种中的表达水平存在显著差异。本研究的表达分析结果,结合已有研究,提示SAD可能参与植物种子生长发育的调控。
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