山杏LACS基因家族生物信息学及表达分析

Bioinformatics and Expression Analysis of the LACS Gene Family in Prunus sibirica

在植物脂肪酸的代谢途径中,长链酰基辅酶A合成酶把游离脂肪酸活化成酰基辅酶A,是调控脂肪酸代谢的重要酶。依据山杏(Prunus sibirica)转录组数据,开展LACS保守域比对分析,共确定6个具有完整ORF的LACS基因,其长度为2 360~2 900 bp,编码氨基酸为654~730 aa。开展LACS蛋白保守域及进化树分析,确定了山杏LACS具有AMP绑定域,说明其在物种间的高度保守性。同时,对不同发育期山杏杏仁的LACS基因开展q RT-PCR表达谱分析。为探究杏仁发育过程中油脂的累积模式,以山杏为试材,采用索氏提取法对其油脂含量进行测定,确定杏仁油的主要累积期在20~50 DAF。结合油脂含量的检测结果,对蛋白互作网络和基因表达等方面加以分析,确定山杏LACS9与油脂累积密切相关。本研究结果可为后续探究LACS基因家族在山杏杏仁发育及油脂累积中的调控机制奠定基础。

In the metabolic pathways of fatty acid in plants, long chain acyl-coenzyme a synthetases （LACSs） activates free fatty acid to acyl-CoA thioesters, which plays an important role in the lipid metabolism. Based on the transcriptomic data of P. sibirica, the homologous analysis on LACS conserved sequence was performed. A total of six LA CSs （the length from 2 360 bp to 2 900 bp） were identified with a completed ORF, which encoded predicted proteins from 654-730 aa. Also, LACS family in P. sibirica was characterized with a conserved AMP binding domain by the analysis of conserved sequence and phylogenetic tree, revealing that it was of high conservation among other species. Meanwhile, expression profiling of LA CS genes at different developmental stages of P. sibirica were carried out by qRT-PCR. In this study, the oil contents in developing kernel of P.sibirica were determined by soxhlet extraction, the result of which showed that the main accumulation period of kernel oil was during 20-50 DAF. According to the detected results of kernel oil, the integrated protein interaction networks and gene expression were analyzed to find that LACS9 in P. sibirica was closely related with the accumulation of oil content. The results of this study could settle the foundation for the further study on the regulatory mechanism of LA CS gene faminly in the almond development and oil accumulation in P. sibirica.