2021年4月23日 星期五
薄荷McHD-Zip3基因的克隆及其调控腺毛发育的功能分析
Cloning of McHD-Zip3 gene of Mentha canadensis and function analysis on its regulatory of development of glandular trichomes
2020年 第29卷 第3期 页码[1-10]
摘要

以薄荷(Mentha canadensis Linn.)cDNA为模版,克隆获得1个HD-Zip转录因子基因,对该基因的组织表达特性以及对转基因烟草(Nicotiana tabacum Linn.)叶片腺毛发育的影响效应进行分析,并对该基因编码的氨基酸序列的特性及其与其他植物的同源性和亲缘关系以及亚细胞定位和转录激活活性进行研究。结果显示:该基因被命名为McHD-Zip3,其编码区(CDS)全长2 226 bp,编码741个氨基酸残基;该基因编码的氨基酸序列的理论相对分子质量为81 375.67,理论等电点为pI 5.92。McHD-Zip3基因编码的氨基酸序列与其他植物HD-Zip转录因子的氨基酸序列具有一定的序列相似性,且均具有保守的Homeodomain、bZip motif、START domain和SAD domain,其与同科植物一串红(Salvia splendens Ker-Gawl.)的SsHD-Zip转录因子的序列相似度最高(82.57%)。在系统树上,McHD-Zip3转录因子与其他植物的多个腺毛发育相关的HD-Zip转录因子聚为一支,同属于HD-Zip Ⅳ亚家族,其与黄花蒿(Artemisia annua Linn.)的AaHD7转录因子的亲缘关系最近。McHD-Zip3转录因子定位于细胞核中,为核定位蛋白,且具有转录激活活性。McHD-Zip3基因的相对表达量在薄荷不同组织中差异显著,其在幼叶和老叶中较高,但在根和茎中较低;McHD-Zip3转基因烟草叶片的腺毛密度明显高于野生型烟草。研究结果表明:McHD-Zip3基因主要在薄荷叶片中表达,McHD-Zip3转录因子属于HD-Zip Ⅳ亚家族,对薄荷腺毛发育具有调控功能。

Abstract

Taking cDNA  of Mentha canadensis Linn. as template,  a HD-Zip transcription factor gene was cloned, and its tissue expression characteristics and effect on development of glandular trichomes on leaf of transgenic Nicotiana tabacum Linn. were analyzed, meanwhile the characteristics of amino acid sequence encoded by this gene, and its homology and genetic relationship with other plants, and subcellular localization and transcriptional activation activity were studied. The results show that this gene is named as McHD-Zip3, and its full length of coding sequence (CDS) is 2 226 bp, which encodes 741 amino acid residues; the theoretical relative molecular mass of amino acid sequence encoded by this gene is 81 375.67, and the theoretical isoelectric point is pI 5.92. There are some sequence similarities in amino acid sequence encoded by McHD-Zip3 gene with those of HD-Zip transcription factors of other plants, and all of them possess conserved Homeodomain, bZip motif, START domain and SAD domain, and its sequence similarity with SsHD-Zip transcription factor of Salvia splendens Ker-Gawl. in the same family is the highest (82.57%). On phylogenetic tree, McHD-Zip3 transcription factor and many HD-Zip transcription factors associated with development of glandular trichomes in other plants are clustered into one branch, all of them belong to HD-Zip Ⅳ subfamily, and its genetic relationship with AaHD7 transcription factor of Artemisia annua Linn. is the closest. McHD-Zip3 transcription factor is a nuclear locating protein localized in cell nucleus, and being with transcriptional activation activity. The relative expression of McHD-Zip3 gene is significantly different in different tissues of M. canadensis, which is relatively high in young leaf and old leaf, but is relatively low in root and stem; the density of glandular trichomes on leaf of McHD-Zip3 transgenic N. tabacum is evidently higher than that of wild type N. tabacum. It is suggested that McHD-Zip3 gene is mainly expressed in leaf of M. canadensis, and McHD-Zip3 transcription factor belongs to HD-Zip Ⅳ subfamily, which has regulatory function on development of glandular trichomes of M. canadensis.

关键词薄荷; McHD-Zip3基因; 表达特性; 亚细胞定位; 转录激活活性; 腺毛发育
Key wordsMentha canadensis Linn.; McHD-Zip3 gene; expression characteristics; subcellular localization; transcriptional activation activity; development of glandular trichome
作者陈泽群, 亓希武, 房海灵, 于盱, 李莉, 梁呈元
所在单位江苏省中国科学院植物研究所(南京中山植物园) 江苏省植物资源研究与利用重点实验室, 江苏 南京 210014
点击量642
基金项目国家自然科学基金资助项目(31970353); 江苏省植物资源研究与利用重点实验室开放基金资助项目(JSPKLB201838)