利用基因芯片技术研究甘蓝型油菜油酸合成中差异表达基因

Differentially Expressed Genes in Oleic Acid Synthesis of Brassica napus by Detected Gene Chip

采用基因芯片技术对甘蓝型油菜高油酸(71.71%)和低油酸(55.6%)材料进行分析,探索油酸的差异表达基因。结果检测到差异表达基因562个,其中上调表达基因194个,下调表达基因368个。以基因芯片中油菜上调基因NM_100489和下调基因NM_130183为材料,用实时荧光定量方法验证基因芯片的结果,二者完全相符。根据基因芯片的实验结果,采用Go注释系统和数据库查询对562个差异表达基因进行功能注释表明,主要为各种酶类、结合功能、转录调控、代谢等,还有的功能未知或与糖代谢及脂肪酸合成相关,其中丙酮酸激酶、果糖二磷酸、酰基传递/酰基ACP硫脂酶、作用于酯键的水解酶、Δ9硬脂酰-乙酰载体蛋白去饱和酶(ADS1)、Δ9酰基-油脂减饱和酶2(ADS2)、ω-3脂肪酸减饱和酶(fad3)等被鉴定为差异表达基因。

Oil from rapeseed with high oleic oil content is insensitive to oxidation and long shelf life.In daily diet with high oleic acid oil can reduce low-density lipoprotein cholesterol levels in blood and prevent arteriosclerosis.In addition,high oleic acid content in edible oil is an important fat indicator.Oleic acid can be effectively converted to methyl ester,which is beneficial to produce biodiesel.Oleic acid content in conventional rapeseed is only 17%.However,oleic acid content in low erucic acid rapeseed reaches around 60%.Differential expressed genes of oleic acid have been found in the high oleic acid line（71.71%） and low oleic acid line（55.6%） of Brassica napus by gene chip technique.The results showed that there were 562 differential expressed genes relating to high and low oleic acid contents in rapeseed,among them 194 were up-regulated genes and 368 were down-regulated genes.The up-regulated gene NM_100489 and the down-regulated gene NM_130183 were then taken as the materials.Real time RT-PCR was further used to confirm the results of gene chip,and the results of real time RT-PCR analysis agreed with the results by gene chip.According to the results of gene chip,562 differentially expressed genes were annotated functionally using Go Annotation System.The molecular function of differential expressed genes mainly included catalysis of enzymes,binding,transcriptional regulation,metabolism and others with unknown functions.Some genes related to sugar metabolism and fatty acid synthesis were identified as differential expressed genes.They are pyruvate kinase,fructose diphosphate,acyl transfer/acyl-ACP S lipase,hydrolase acting on ester bond,Δ9 stearoyl acid-N-carrier protein desaturase（ADS1）,Δ9 acyl-lipid desaturase 2（ADS2）,ω-3 fatty acid desaturation enzymes（fad3） and so on.