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百合LhTPS基因启动子的克隆及序列分析
韩婷,王婷,李兴桃,杜方
0
(山西农业大学 园艺学院, 山西 太谷 030800)
摘要:
为明确百合LhTPS基因的启动子序列特征,利用反向PCR技术和DNAMAN、PlantCARE等分析软件,对8个不同百合品种萜烯合酶基因LhTPS的启动子进行克隆和生物信息分析。结果表明:1)克隆得到的LhTPS基因启动子序列长度在1 376~1 414 bp,相似性为90.26%,核苷酸多态性位点404个,大于5 bp的插入缺失有6处;2)不同百合品种的LhTPS启动子序列均包括核心元件和应答元件,应答元件又分为生长发育、光响应、胁迫响应及激素响应4类,其中与MeJA反应相关的顺式作用元件(CGTCA-motif、TGACG-motif、MYC和MYB)所占比例最大;3)根据启动子序列信息,可将8个百合品种分为4组,即东方百合组、亚洲百合组、喇叭百合组和东喇百合组,分类结果与传统分类结果一致。综上,LhTPS基因启动子序列的首次获得,为日后进行LhTPS启动子活性和功能分析奠定了基础,也为阐明百合萜类挥发物形成的调控机制奠定了理论基础。
关键词:  百合  萜烯合酶  启动子  顺式作用元件  花香
DOI:10.11841/j.issn.1007-4333.2022.05.17
投稿时间:2021-09-09
基金项目:山西农业大学科技创新基金项目(2019yz005)、山西省基础研究计划(自由探索类)项目(20210302123416)
Cloning and sequence analysis of LhTPS gene promoter in lilies
HAN Ting,WANG Ting,LI Xingtao,DU Fang
(College of Horticulture, Shanxi Agricultural University, Taigu 030801, China)
Abstract:
In order to clarify the promoter sequence characteristics of terpene synthase gene(LhTPS)in lily, the promoters of eight lily cultivars were cloned and bioinformatically analyzed using inverse PCR technique and bioinformatic software such as DNAMAN and PlantCARE. The results showed that: 1)The cloned LhTPS gene promoter sequences were 1 376-1 414 bp in length. The homology of gene promoters was 90. 26%. A total of 404 nucleotide polymorphism sites and 6 indels larger than 5 bp were identified; 2)There were core elements and responsiveness elements in LhTPS promoters. The responsiveness elements were classified into into 4 categories, which were plant growth and development responsiveness, light responsiveness, stress responsiveness and phytohormone, responsiveness, respectively. CGTCA-motif, TGACG-motif, MYC and MYB associated with MeJA-responsiveness and contained the largest proportion of cis-element accounting; 3)Based on the promoter sequence information, the eight lily cultivars were divided into four groups, e. g. , Oriental, Asiatic, Trumpet and Orienpet, and it was consisted with the traditional classification. In conclusion, research on the first cloned LhTPS promoters laid a basis for future studies on their activity and function, and would also be beneficial for elucidation of regulatory mechanism of lily terpenoids.
Key words:  Lilium  terpene synthase  promoter  cis-acting elements  floral scent