两个非洲菊4CL基因的克隆及表达  

Cloning and expression analysis of two 4CL genes in gerbera

作　　者：王星淇 孙雪丽 车婧如 石小保 程春振[1] WANG Xingqi;SUN Xueli;CHE Jingru;SHI Xiaobao;CHENG Chunzhen(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农林大学园艺学院,福州350002

出　　处：《应用与环境生物学报》2020年第2期272-279,共8页Chinese Journal of Applied and Environmental Biology

基　　金：福建省高原学科建设经费(102/71201801101);福建农林大学“校杰出青年科研人才”计划项目(xjq201721)资助

摘　　要：香豆酸:辅酶A连接酶(4CL)是苯丙烷类代谢的一个关键酶,在植物生长发育和抗逆防御反应中发挥着重要作用.为了解非洲菊4CL基因的序列特征和表达特性,使用反转录PCR克隆两个非洲菊4CL基因,利用生物信息学分析软件分析它们的序列特性,同时利用qRT-PCR研究它们在不同组织部位和低温、干旱、盐胁迫和水杨酸(SA)处理下的表达情况.结果显示,两个非洲菊4CL基因(Gj4CL-like7和Gj4CL-like1)CDS长度分别为1 617 bp和1 623 bp,它们编码的蛋白均为疏水蛋白,其中Gj4CL-like7拥有跨膜结构.Gj4CL含有木质素代谢相关4CL特有保守序列SSGTTGLPKGV和GEICIRG.定量结果显示,它们在不同组织部位中的表达情况相同均为根>叶柄>叶;低温处理下Gj4CL-like7的表达先升高,在处理8 h时达到最大值,之后显著下调并显著低于对照,而Gj4CL-like1的表达呈"升—降—升—降"的趋势,低温处理前12 h的表达量均高于对照且在12 h时最高,之后显著低于对照;盐胁迫下,Gj4CL-like7的表达受到显著抑制,而Gj4CL-like1的表达早期呈下调趋势,处理24h后的表达量显著高于对照;干旱胁迫和SA处理下这两个基因的表达均受到显著抑制,且Gj4CL-like7受抑制程度更高.本研究表明Gj4CL-like1主要参与木质素代谢,在非洲菊逆境胁迫应答中发挥着重要作用;Gj4CL-like7编码蛋白具有4CL保守序列相似序列,可能参与类黄酮代谢.(图6表1参36)One of the key enzymes in the phenylpropanoid pathway,4-coumarate:coenzyme A ligase(4CL),plays an important role in plant growth and defense.To investigate the sequence and expression characteristics of Gerbera 4CL genes,two Gerbera 4CL genes were amplified using reverse transcription PCR.Then,the sequences were subjected to bioinformatics analysis.Their expression patterns in different tissues or organs as well as under different abiotic stress conditions(e.g.,cold,drought,salt,and salicylic acid(SA)),were also investigated.Two gerbera 4CL genes,named Gj4CL-like7 and Gj4CL-like1,were successfully cloned,and the length of their coding sequences were 1617 bp and 1623 bp,respectively.Both their encoded proteins were hydrophobic,and transmembrane structure was found in Gj4CL-like7 but not in Gj4CL-like1.Moreover,Gj4CLlike1 contains two unique conserved sequences,SSGTTGLPKGV and GEICIRG,of the lignin metabolismrelated 4CLs.Quantitative real time PCR(qRT-PCR)showed that the expression of both 4CLs followed the order of root>petiole>leaf.In the low temperature treatment,the expression of Gj4CL-like7 was significantly induced in the early stages and peaked at 8 h,after which its expression was found to be significantly inhibited.The expression of Gj4CL-like1 showed an‘up-down-up-down’pattern,and its expression in the three early stages was higher than that of the control.The highest expression was found at 12 h;however,its expression was found to be significantly lower than that of the control at 24 h.In the drought and SA treatments,the expression of both Gj4CLs was significantly inhibited,and the inhibitory effect of drought on Gj4CL-like7 was the most severe.In the salt treatment,the expression of Gj4CL-like7 was greatly inhibited at all stages,while the expression of Gj4CL-like1 was significantly higher than that of the control after a 24-h salt treatment,although its expression was also inhibited in the early stages.It was predicted that Gj4CL-like1 participates in the lignin metabolism and stress responses of

关 键 词：非洲菊 4-香豆酸:辅酶A连接酶(4CL) 木质素 类黄酮 基因表达 

分 类 号：S682.11[农业科学—观赏园艺] Q943.2[农业科学—园艺学]