摘要
HD-Zip是植物界一类特有的转录因子,影响植物的生长发育及次生代谢。其中的HD-ZipⅣ被认为在毛状体的发育中具有重要的调控作用。为了探究荆芥HD-ZipⅣ亚家族的2个基因StHD1与StHD8在荆芥腺鳞中的调控作用,利用转录组测序技术、酵母双杂技术等对2个基因的表达模式、蛋白之间的互作进行了分析,并对基因进行了亚细胞定位、在烟草体内和荆芥体内的功能验证。结果表明:荆芥的StHD1与StHD8与其他植物的HD-ZipⅣ亚家族蛋白序列相似性较高,具有HD-ZipⅣ亚家族特征保守结构域,亚细胞定位显示被定位于细胞核;以上2个基因主要在幼嫩组织和花穗中表达;酵母双杂结果表明StHD1与StHD8蛋白之间存在互作。StHD1与StHD8过表达的转基因烟草腺毛密度和长度明显增加。通过VIGS(virus-induced gene silencing)技术在荆芥体内抑制StHD1与StHD8表达会导致腺鳞密度、单萜合成关键基因表达量下降,单萜主要成分柠檬烯和胡薄荷酮相对含量降低。以上结果表明,荆芥的StHD1与StHD8通过形成复合体对荆芥的腺毛具有调控作用,同时影响单萜的生物合成。
Hd-Zip, a unique transcription factor in plant kingdom, influences the growth, development, and secondary metabolism of plants. Hd-zip Ⅳ is thought to play an important role in trichome development of Schizonepeta tenuifolia. This study aims to explore the functions of StHD1 and StHD8 in Hd-zip Ⅳ subfamily in peltate glandular trichome development. To be specific, the expression patterns of the two genes and interaction between the proteins encoded by them were analyzed based on transcriptome sequencing and two-hybrid screening. The subcellular localization was performed and functions of the genes were verified in tobacco and S. tenuifolia. The results showed that StHD1 and StHD8 had high similarity to HD-Zip Ⅳ proteins of other plants and they all had the characteristic conserved domains of HD-Zip Ⅳ subfamily. They were located in the nucleus. The two genes mainly expressed in young tissues and spikes, and StHD1 and StHD8 proteins interacted with each other. The density and length of glandular trichomes increased significantly in tobacco plants with the overexpression of StHD1 and StHD8. Inhibiting the expression of StHD1 and StHD8 by VIGS(virus-induced gene silencing) in S. tenuifolia resulted in the reduction in the density of peltate glandular trichomes, the expression of key genes related to mono-terpene synthesis, and the relative content of limonene and pulegone, the main components of monoterpene. These results suggested that StHD1 and StHD8 of S. tenuifolia formed a complex to regulate glandular trichomes and affect the biosynthesis of monoterpenes.
作者
周佩娜
党静洁
邵永芳
石遵睿
张琳
刘潺潺
吴啟南
ZHOU Pei-na;DANG Jing-jie;SHAO Yong-fang;SHI Zun-rui;ZHANG Lin;LIU Chan-chan;WU Qi-nan(School of Pharmacy,Nanjing Universily of Chinese Medicine,Nanjing 210023,China;Collaborative Innovation Center of Chinese Medicinal Resources Indlustrialization,Nanjing 210023,China;National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Inovativre Medicine,Nanjing 210023,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2022年第21期5838-5848,共11页
China Journal of Chinese Materia Medica
基金
国家自然科学基金面上项目(81973435)
国家自然科学基金青年基金项目(81903756)
江苏省研究生科研与实践创新计划项目(KYCX21_1759,KYCX22_2031)。
关键词
荆芥
VIGS
蛋白互作
转基因
qPCR
Schizonepeta tenuifolia
VIGS
protein-protein interaction
transgenic
qPCR
