一个与氰苷生物合成相关的植物CYP基因的克隆与表达分析

Cloning and Expression Analysis of a Plant Cytochrome P450 Gene Involved in the Biosynthesis of Cyanogenic Glycoside

氰苷是由植物细胞色素P450单加氧酶(CYP)催化氨基酸生成的次生代谢产物,与植物的防御和抗逆境胁迫相关。为了获得蒜头果中与氰苷生物合成有关的CYP基因,本研究通过分析蒜头果转录组数据,从中分离并克隆得到1条细胞色素P450基因(命名为MoCYP79),并对其进行生物信息学分析及检测该基因在果实不同发育时期的具体表达情况。结果显示,MoCYP79基因的cDNA全长1632 bp,编码543个氨基酸;MoCYP79蛋白含有CYP家族的识别结构域血红素结合域(FSTGRRGC),且与巨桉(XP_010027351.1)、木薯(AAF27290.1,AAF27289.1)等植物CYP79蛋白聚在一支;MoCYP79基因在蒜头果花谢后的果实中表达量呈下降趋势,除发育3个月的果实外,该基因在其余的果实中的表达量均显著高于叶片,从大到小依次为花谢后的果实1个月>2个月>4个月3个月。本实验对研究蒜头果的对病虫害的防御、逆境胁迫的抵御及蒜头果中有效次生代谢产物的发掘具有重要的意义。

Cyanogenic glycoside is a secondary plant metabolism product,which derived from the conversion of amino acid catalyzing by the cytochrome P450 monooxygenase(CYP),related to plants’defense and stress resistance.In order to obtain the CYP gene corresponding to the biosynthesis of cyanogenic glycosides,the present study isolated and cloned a cytochrome P450 gene(named Mo CYP79)through analyzing the transcriptome of Malania oleifera,simultaneously,carried on bioinformatic analysis of MoCYP79 and detected the relative expression this gene during different developmental period.The results showed that the full length c DNA of MoCYP79 has1632 bp,codes 543 amino acids.The protein MoCYP79 contains the identification characters heme-binding domain(FSTGRRGC)of CYP79 family,the protein and Eucalyptus grandis(XP010027351.1),Manihot esculenta(AAF27290.1,AAF27289.1)clustered one clan.The expression level of MoCYP79 in the fruit of M.oleifera after blossom fading occurred downward trend,the expression level of this gene was higher than other developmental period of the fruit except 3 months fruit,the expression level was 1 st month fruit>2 nd month fruit>4 th month fruit successively.The experiment is of great significance to study the defense from the disease and insect damaging,anti-envirommental stress and the exploration of secondary metabolism in M.oleifera.