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[1]张璐 吴菁华** 姚馨婷 王革伏 张志忠**.中国水仙NtTFL1-1和NtTFL1-2基因的克隆和表达*[J].应用与环境生物学报,2020,26(02):1-9.[doi:10.19675/j.cnki.1006-687x.2019.06007]
 ZHANG Lu,WU Jinghua**,YAO Xinting,et al.Cloning and expression of NtTFL1-1 and NtTFL1-2 genes from Narcissus tazetta var. chinensis*[J].Chinese Journal of Applied & Environmental Biology,2020,26(02):1-9.[doi:10.19675/j.cnki.1006-687x.2019.06007]
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中国水仙NtTFL1-1和NtTFL1-2基因的克隆和表达*()
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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02期
页码:
1-9
栏目:
研究论文
出版日期:
2020-03-20

文章信息/Info

Title:
Cloning and expression of NtTFL1-1 and NtTFL1-2 genes from Narcissus tazetta var. chinensis*
作者:
张璐1 吴菁华1** 姚馨婷1 王革伏12 张志忠1**
1福建农林大学园艺学院,福建农林大学-戴尔豪西联合实验室 福州 350002
2加拿大戴尔豪西大学农学院 加拿大特鲁罗 N2B 5E3
Author(s):
ZHANG Lu1 WU Jinghua1** YAO Xinting1 Gefu WANG-PRUSKI 1 2 & ZHANG Zhizhong 1**
1 Joint FAFU-Dalhousie Lab, College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, 350002, China
2 Faculty of Agriculture, Dalhousie University, Truro, B2N 5E3, Canada
关键词:
中国水仙TFL1基因花芽分化荧光定量PCR亚细胞定位
Keywords:
Narcissus tazetta var. chinensis TFL1 gene flower bud differentiation qPCR subcellular localization
DOI:
10.19675/j.cnki.1006-687x.2019.06007
摘要:
中国水仙(Narcissus tazetta var. chinensis)是十大传统名花之一,只在冬季开花,为典型的年宵花卉,开花时间的局限使其应用价值受到一定限制。水仙花球在夏季的休眠期进行花芽分化,是花期调控研究的重点阶段,但目前对其开花机制和调控的研究尚处于起步阶段。TFL1(TERMINAL FLOWER 1)是植物重要的开花相关基因,为了解该基因在中国水仙花芽分化和开花过程中的作用,以中国水仙‘金盏银台’为材料,利用逆转录聚合酶链式反应(RT-PCR,Reverse Transcription-Polymerase Chain Reaction)技术克隆了中国水仙的TFL1基因:NtTFL1-1和NtTFL1-2,对获得的基因序列进行了相关生物信息学分析,对该基因编码蛋白进行了亚细胞定位;利用荧光定量PCR技术对其在中国水仙花芽分化不同时期、不同器官类型中的差异表达情况进行了测定。结果显示NtTFL1-1开放阅读框(ORF,Open Reading Frame)共498碱基对(bp,base pair),编码166个氨基酸,NtTFL1-2的ORF共522 bp,编码174个氨基酸;二者均具有TFL1家族的保守结构域;NtTFL1-1和NtTFL1-2均定位在细胞质中。NtTFL1-1和NtTFL1-2在中国水仙花芽分化初期和花序原基形成期的表达量较低,在花原基形成期表达量较高;NtTFL1-1和NtTFL1-2在根中的表达量高于茎、叶和成熟的花。本研究结果显示NtTFL1-1和NtTFL1-2和中国水仙花芽分化密切相关,实验结果可为中国水仙花期调控研究提供理论参考。(图6 表2 参30)
Abstract:
Objectives: Narcissus tazetta var. chinensis ( Nt) is one of the ten famous traditional flowers. It blossoms only in winter and is a typical Spring Festival flowers. The fixed florescence limits its application value. The flower bud differentiation of Nt occurs in the dormancy period of the bulb in summer. This period is the key stage of the research of flowering time regulator. However, the study on flowering mechanism and regulation is still in its infancy. TFL1 is an important flowering-related gene in plants. In order to reveal the role of TFL1 gene in flower bud differentiation and flowering of Nt, we cloned the TFL1 gene from Nt and analyzed its related bioinformatics. TFL1 protein subcellular localization and TFL1 gene expression level were analyzed. The results will be helpful to provide references for florescence regulation and breeding of different florescence Nt varieties. Methods: T he TFL1 gene of Nt were cloned by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) using Nt cv. ‘ Jinzhanyintai’ as material. Two clones were obtained in this experiment and named NtTFL1-1 and NtTFL1-2 . The obtained gene sequence was analyzed by bioinformatics, and the subcellular localization of the gene encoding protein was carried out. The differential expression of the gene in different stages of flower bud differentiation and different organ types of Nt was determined by qPCR. Results: Bioinformatics analysis showed that the ORF of NtTFL1-1 was 498 bp, encoding 166 amino acids, and the ORF of NtTFL1-2 was 522 bp, encoding 174 amino acids, all of which had conserved domains of the TFL1 family. NtTFL1-1 and NtTFL1-2 were localized in the cytoplasm. The results of qRT-PCR analysis showed that the expression of NtTFL1-1 and NtTFL1-2 genes was lower in the early stage of flower bud differentiation and inflorescence primordium formation and higher in flower primordium formation. The expression of NtTFL1-1 and NtTFL1-2 in roots was significantly higher than that in stems, leaves and mature flowers. Conclusions: The results confirmed that NtTFL1-1 and NtTFL1-2 expressed differently in the flower bud differentiation stage of Nt, which were closely related to the flower bud differentiation of Nt. The experimental results could provide theoretical reference for flower bud regulation of Nt.

相似文献/References:

[1]姚红,周平,范雨昕,等.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):993.[doi:10.19675/j.cnki.1006-687x.2018.10009]
 YAO Hong,ZHOU Ping,FAN yuxin,et al.Cloning and functional analysis of DFR gene promoter in Narcissus tazetta var. chinensis[J].Chinese Journal of Applied & Environmental Biology,2019,25(02):993.[doi:10.19675/j.cnki.1006-687x.2018.10009]

备注/Memo

备注/Memo:
收稿日期 Received: 2019-06-05 接受日期 Accepted: 2019-08-16
*国家自然科学基金项目(31601783)、福建省级扶贫开发工作重点县人才支持计划专项(K1519060A)和福建农林大学创新基金项目(CXZX2017533)资助
**通讯作者 Corresponding author (E-mail: wjhham@163.com; zeada2001@163.com)
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更新日期/Last Update: 2019-09-04