摘要
【目的】探讨山核桃中oleosin基因家族表达模式,筛选响应油脂合成的关键oleosin基因。【方法】以山核桃(Carya cathayensis)和薄壳山核桃(Carya illinoinensis)为研究对象,利用生物信息学方法鉴定oleosin基因家族成员,分析其蛋白的理化性质、保守基序;解析基因结构、启动子区域顺式作用元件;预测oleosin基因的系统进化关系;探究oleosin基因在胚发育不同阶段的表达模式及亚细胞定位。【结果】山核桃和薄壳山核桃基因组中分别鉴定到7个和8个含“脯氨酸结”保守基序PX5SPX3P的oleosin基因家族成员,这些基因所编码的蛋白分子质量14.54~17.51 ku;启动子区域顺式作用元件分析显示,oleosin基因家族成员均含有响应光周期、逆境胁迫和激素应答等相关的顺式作用元件;系统进化分析显示15个oleosin基因家族成员主要分布在U-Oleosin、SL-Oleosin和SH-Oleosin 3个亚家族;转录组数据显示,山核桃7个oleosin基因在胚发育不同阶段均表现出不同程度上调表达,其中CCA0520S0025和CCA0791S0002在山核桃胚油脂迅速增长阶段基因表达量显著上调;荧光定量PCR结果显示,山核桃oleosin基因CCA0520S0025和CCA0791S0002响应ABA和低温胁迫,同时亚细胞定位分析表明山核桃CCA0520S0025和CCA0791S0002蛋白均定位于油体。【结论】山核桃和薄壳山核桃oleosin基因家族成员均具有相似的基因结构与保守基序,亲缘关系较近,另外山核桃CCA0520S0025和CCA0791S0002可能在油脂合成过程中发挥重要作用。
【Objective】Lipid droplet(LD)plays an essential role in seed germination.Oil body proteins(oleosin)are a kind of proteins localized on the surface of LD,which participates in the regulation of LD biogenesis,morphology homeostasis,and stabilization.Chinese hickory(Carya cathayensis)is an arbor with high oil content in its embryo.However,the molecular mechanism of high oil accumulation in hickory embryos remains unclear.This study aimed to systematically investigate the oleosin genes in the Chinese hickory and analyze their expression patterns.【Methods】Based on the Pfam database,the oleosin genes in Chinese hickory(C.cathayensis)and pecan(C.illinoinensis)were identified and characterized in this study.Then,gene structures and conserved motifs were analyzed by TBtools,and the cis-acting elements of oleosin gene promoter regions were performed using the Plant CARE database,and phylogenetic analysis of oleosin gene family from C.cathayensis,C.illinoinensis,Arabidopsis thaliana,Glycine max,Arachis hypogaea and Brassica napuswere conducted using MEGA 7.0 software(Bootstrap=1000).Moreover,the expression patterns during different developmental stages of the embryo were analyzed.Furthermore,subcellular localization of oleosin proteins was conducted.【Results】The results showed that seven and eight oleosin genes containing a conserved domain“proline knot”were identified in Chinese hickory and pecan,respectively.The oleosin protein molecular weights ranged from 14.54 to 17.51 ku,and the isoelectric point of oleosin protein ranged from 9.33 to 10.36.The gene structure analysis showed that all members of the oleosin genes in hickory and pecan contained one exon but not intron.Four conserved motifs were found in the oleosin gene family.Based on the analysis of the conserved motifs,CCA0791S0002,CIL0938S0150 and CIL0252S0009 might share a similar function.The results of multiple amino acid sequence alignment showed that oleosin proteins of hickory and pecan contained a typical and highly conserved“proline knot”-
作者
董茗秀
易芳
胡帅
高燕丽
DONG Mingxiu;YI Fang;HU Shuai;GAO Yanli(Zhejiang A&F University/State Key Laboratory of Subtropical Silviculture,Lin’an 311300,Zhejiang,China)
出处
《果树学报》
CAS
CSCD
北大核心
2023年第1期35-47,共13页
Journal of Fruit Science
基金
浙江农林大学科研发展基金(2021FR012,2020KJ001)。
