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[1]王星淇 孙雪丽 车婧如 石小保 程春振**.两个非洲菊4CL基因的克隆及表达[J].应用与环境生物学报,2020,26(03):1-12.[doi:10.19675/j.cnki.1006-687x.2019.07025]
 WANG Xingqi,SUN Xueli,CHE Jingru,et al.Cloning and expression analysis of two 4CL genes in Gerbera[J].Chinese Journal of Applied & Environmental Biology,2020,26(03):1-12.[doi:10.19675/j.cnki.1006-687x.2019.07025]
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两个非洲菊4CL基因的克隆及表达()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
26卷
期数:
2020年03期
页码:
1-12
栏目:
研究论文
出版日期:
2020-06-25

文章信息/Info

Title:
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**
福建农林大学园艺学院 福州 3 0002
Author(s):
WANG Xingqi SUN Xueli CHE Jingru SHI Xiaobao& CHENG Chunzhen**
College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China
关键词:
非洲菊4-香豆酸:辅酶A连接酶(4CL)木质素类黄酮表达
Keywords:
Gerbera 4-coumarate: coenzyme A ligase (4CL) lignin flavonoid expression analysis
DOI:
10.19675/j.cnki.1006-687x.2019.07025
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。为研究非洲菊4CL基因的序列特征和表达特性,使用反转录PCR克隆了两个非洲菊4CL基因,利用生物信息学分析软件分析了它们的序列特性,同时利用qRT-PCR研究了它们在不同组织部位和低温、干旱、盐胁迫和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1617 bp和1623 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24 h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)
Abstract:
The 4-coumarate: coenzyme A ligase ( 4CL), one of the key enzymes in the phenylpropanoid pathway, plays important roles in plant growth and defense responses. 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 series of bioinformatics analysis. Their expression patterns in different tissues or organs, and under different abiotic stresses, such as cold, drought, salt and SA treatments, were also investigated . Two gerbera 4CL genes (respectively named as Gj4CL-like7 and Gj4CL-like1) were successfully cloned, and the length of their coding sequence was respectively 1617 bp and 1623 bp. Both their encoded proteins are hydrophobic, and transmembrane structure was found in Gj4CL-like7 protein but not in Gj4CL-like1. Moreover, Gj4CL-like1 contains two unique conserved sequences (SSGTTGLPKGV and GEICIRG) of the lignin metabolism related 4CLs. Quantitative real time PCR ( qRT-PCR) result showed that both the two 4CLs’ expression follows the order: root > petiole > leaf. Under low temperature treatment, the expression of Gj4CL-like7 was significantly induced in the early stages and peaked at 8 h, then its expression was found to be significantly inhibited. While the expression of Gj4CL-like1 showed a ‘up-down-up-down’ pattern, and its expression in the three early stages were all higher than control and it highest expression was found at 12 h, at 24 h, however, its expression was found to be significantly lower than the control. Under drought and SA treatments , both the two Gj4CLs’ expression was significantly inhibited, and the inhibitory effect of drought on Gj4CL-like7 was severer. Under salt treatment, the expression of Gj4CL-like7 was greatly inhibited at all the stages, while the expression of Gj4CL-like1 was significantly higher than control after 24 h’s salt treatment although its expression was also inhibited at the early stages. It was predicted that Gj4CL-like1 participates in the lignin metabolism and stress response s of gerbera. Gj4CL-like7 encoded protein contains similar conserved sequences of 4CL, and it was predicted to play roles in the flavonoid metabolism.

备注/Memo

备注/Memo:
收稿日期 Received: 2019-07-14 接受日期 Accepted: 2019-11-15
*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
**通讯作者 Corresponding author (E-mail: ld0532cheng@126.com)
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更新日期/Last Update: 2019-12-04