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[1]姚 红 周 平 范雨昕 孙瑞琦 Muhammad Anwar 曾黎辉**.中国水仙DFR基因启动子的克隆及功能*[J].应用与环境生物学报,2019,25(04):1-9.[doi:10.19675/j.cnki.1006-687x.2018.10009]
 YAO Hong,ZHOU Ping,FAN yuxin,et al.Cloning and Functional Analysis of Promotor of DFR gene in Narcissus tazetta var. Chinensis*[J].Chinese Journal of Applied & Environmental Biology,2019,25(04):1-9.[doi:10.19675/j.cnki.1006-687x.2018.10009]
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中国水仙DFR基因启动子的克隆及功能*()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年04期
页码:
1-9
栏目:
简报
出版日期:
2019-07-30

文章信息/Info

Title:
Cloning and Functional Analysis of Promotor of DFR gene in Narcissus tazetta var. Chinensis*
文章编号:
201810009
作者:
姚 红 周 平 范雨昕 孙瑞琦 Muhammad Anwar 曾黎辉**
福建农林大学园艺学院 福州 350002
Author(s):
YAO Hong ZHOU Ping FAN yuxin SUN ruiqi MUHAMMAD Anwar&ZENG Lihui**
College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002
关键词:
中国水仙二氢黄酮醇4-还原酶基因启动子功能鉴定
Keywords:
Narcissus tazetta var. Chinensis dihydroflavonol 4-reductase gene promoter functional identification
DOI:
10.19675/j.cnki.1006-687x.2018.10009
摘要:
为了解中国水仙花青素合成途径关键基因DFR(NtDFR)的表达调控以及中国水仙不能合成花青素的分子机制,本研究采用染色体步移法从中国水仙中克隆了NtDFR基因起始密码子上游962 bp的启动子序列。生物信息学分析结果表明,启动子序列除包含TATA-box、CAAT-box等基本启动子元件外,还包含光调控元件、植物激素响应元件、胁迫响应元件等多个顺式作用元件。此外,该启动子序列还含有MYB转录因子结合位点。为验证启动子的表达特性,将NtDFR启动子取代植物表达载体pBI121上35S启动子,构建pBI121-pNtDFR::GUS载体,利用农杆菌转化烟草叶片瞬时表达,通过GUS组织染色法确定了克隆的启动子的活性。将中国水仙R2R3-MYB转录因子NtMYB2、NtMYB5分别和pBI121-pNtDFR::GUS共同注射烟草,定量PCR和GUS组织化学染色结果表明NtMYB2和NtMYB5 都使NtDFR启动子诱导的烟草叶片GUS颜色变浅以及GUS基因表达量下降,表明NtMYB2和NtMYB5是NtDFR的抑制因子。研究结果有助于了解中国水仙花青素合成途径的分子调控机制。
Abstract:
Objectives: To understand the expression regulation of dihydroflavonol 4-reductase gene (NtDFR), the key gene in the pathway of anthocyanin biosynthsis in Chinese narcissus (Narcissus tazetta var. chinensis) and the molecular mechanism of no anthocyanin accumulation in Chinese narcissus.Methods: The 962 bp promoter fragment of NtDFR gene was cloned from genomic DNA by genome walking method. The plant expression vector pBI121 was used to construct the pBI121-pNtDFR::GUS vector. The activity of promoter was determined by GUS histochemical staining with agro-infiltrated tobacco leaves. Results: The results of promoter sequence analysis show that the promoter contains a number of TATA-box, CAAT-box elements and other promoter elements, such as light responsive elements, hormone response elements, stress-response elements etc. Moreover, the promoter also contains the MYB transcription factor combination elements. In addition, NtMYB2 and NtMYB5, R2R3-MYB genes of Chinese narcissus, were transient expression with pBI121-pNtDFR::GUS in tobacco leaves seperately. The results of real-time quantitative PCR and GUS histochemical staining analysis indicate that NtMYB2 and NtMYB5 repress the activity of the NtDFR promoter. Conclusions: NtMYB2 and NtMYB5are the repressors of NtDFR.? ? ? ? ? ? ?

备注/Memo

备注/Memo:
收稿日期:2018-10-09 接受日期 Accepted: 2018-11-29*福建农林大学科技创新基金(KFA17352A,KFA17602A)和福建农林大学国际科技合作与交流项目(Kxb16013A)资助 **通讯作者(E-mail: lhzeng@hotmail.com)点击摘要页题目后的“PDF”可下载阅读全文;本文为已录用的作者修定稿,尚未经编辑全面修改。引用本文请注明出处本刊;发表刊期和页码将以正式出版时的安排为准,但DOI确定不变。
更新日期/Last Update: 2018-12-28