鸭儿芹肉桂酸4-羟化酶基因的克隆与不同温度下的表达分析

Isolation and Expression Analysis of Cinnamic Acid 4-hydroxylase Gene under Different Temperatures in Cryptotaenia japonica Hassk

为研究鸭儿芹木质素合成相关基因,以贵州省野生鸭儿芹为研究对象,克隆到1条1 631bp长的肉桂酸4-羟化酶(C4H)基因(CjC4 H),并以鸭儿芹生长过程中主要面临的4种不同温度(10℃、18℃、30℃和38℃)处理0、0.5、1、2、4、8、12和24h,考察鸭儿芹木质素合成基因CjC4 H的表达情况,探讨鸭儿芹木质素合成基因受温度的调控关系,为鸭儿芹适宜生长温度的选择提供参考。开放阅读框(ORF)分析显示,CjC4 H基因的开放阅读框(ORF)长度为1 521bp,可编码506个氨基酸,蛋白分子质量为58.13kD,等电点为9.38。进化树分析表明,同科植物之间C4H进化上相对保守。实时定量PCR检测鸭儿芹中CjC4 H基因与温度的响应表达,结果表明38℃高温处理4h时,CjC4 H基因表达量远高于其他温度处理;而10℃和18℃相对低温处理,CjC4 H基因表达在24h时明显提高。

In order to study the lignin synthesis related genes of Cryptotaenia japonica Hassk,the gene CjC4H with 1 631 bp,which encoding the cinnamate 4-hydroxylase（C4H） involved into lignin synthesis,was cloned from wild variety C.japonica Hassk of Guizhou Province.C.japonica Hassk was grown under 4 different temperatures（10 ℃,18 ℃,30 ℃ and 38 ℃）.The CjC4H genes expression was detected at 0,0.5,1,2,4,8,12 and 24 h to provide reference for C.japonica Hassk suitable growth temperature selec tion.The ORF（Open Reading Frame） length was 1 521 bp,which encoding 506 amino acids.The molecular weight of the enzyme of CjC4H was 58.13 kD,and the pI was 9.38.Phylogenetic analysis demonstrated that the enzyme of C4H relative conservative in different plant species.The CjC4H genes expression under different temperature detected by real time quantitative PCR,the results showed that the CjC4H gene expression under high temperature of 38 ℃ 4 h was much higher than that of other temperature treatments,and the expression of CjC4H increased significantly at 24 h under 10 ℃ and 18 ℃ low temperature treatments.