| 引用本文: | 黄小龙, 陈婷婷, 张琴琴, 莫佳佳, 龚盼琴, 闫慧清.刺梨GL2同源基因的克隆、系谱树和表达分析[J].广西植物,2020,40(1):119-127.[点击复制] |
| HUANG Xiaolong, CHEN Tingting, ZHANG Qinqin,
MO Jiajia, GONG Panqin, YAN Huiqing.Cloning, phylogenic and expression analysis of GL2 homology gene in Rosa roxburghii[J].Guihaia,2020,40(1):119-127.[点击复制] |
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| 刺梨GL2同源基因的克隆、系谱树和表达分析 |
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黄小龙1,2, 陈婷婷1,2, 张琴琴1,2, 莫佳佳1,2, 龚盼琴1,2, 闫慧清1*
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1. 贵州师范大学 生命科学学院, 贵州省植物生理与发育调控重点实验室, 贵阳 550001;2. 国家林业局西南喀斯特山地生物多样性保护重点实验室, 贵阳 550001
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| 摘要: |
| 为了观察刺梨果实的果刺细胞学发育过程,该研究以刺梨‘贵农5号'的cDNA为模板,通过RACE克隆获得刺梨中与拟南芥表皮毛形成GL2的同源基因RrGL2,并对该基因进行生物信息学分析和表达分析。结果表明:(1)刺结构在花芽形成早期基部内的细胞首先不断分裂,向外继续发育,中部的细胞变细、变长形成“针”状结构,顶部的细胞逐渐木质化使刺变硬,形成果刺。(2)通过RACE扩增得到RrGL2的cDNA全长2 292 bp,编码763 aa氨基酸。(3)RrGL2具有Homeodomain同源结构域和StAR磷脂酰胆碱转移蛋白的结构域,RrGL2与其他物种编码的GL2氨基酸同源性高度相似,并且系谱树分析揭示刺梨RrGL2和野草莓的GL2密切相关。(4)qRT-PCR分析表明,RrGL2在茎和果实中的表达水平高于其他组织,在花后7周果刺中的表达最高,是3周和5周果刺中的7.87倍和2.10倍。综上结果发现RrGL2的功能与果刺的形成发育密切相关,该研究为刺梨中刺形成的分子机制和育种提供了理论基础。 |
| 关键词: 刺梨, 表皮毛, 果刺, RrGL2, 基因表达 |
| DOI：10.11931/guihaia.gxzw201810025 |
| 分类号:Q949.45 |
| 文章编号:1000-3142(2020)01-0119-09 |
| 基金项目:国家自然科学基金(31660554); 贵州省科学技术基金(黔科合J字[2015]2117号)[Supported by the National Natural Science Foundation of China(31660554); Guizhou Science and Technology Fund([2015]2117)]。 |
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| Cloning, phylogenic and expression analysis of GL2 homology gene in Rosa roxburghii |
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HUANG Xiaolong1,2, CHEN Tingting1,2, ZHANG Qinqin1,2,
MO Jiajia1,2, GONG Panqin1,2, YAN Huiqing1*
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1. Key Laboratory of Plant Physiology and Development Regulation, School of Life Sciences, Guizhou Normal University, Guiyang 550001, China;2. National Forestry Administration, Karst Key Laboratory of Biodiversity Conservation in Southwest China, Guiyang 550001, China
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| Abstract: |
| In order to observe cytological development of prickles in fruits of Rosa roxburghii, R. roxburghii GLABROUS 2(RrGL2), a prickle-development related AtGL2 homology gene, was isolated from ‘Guinong 5' and relative biological information and expression were analyzed in this paper. The cytological development of fruit thorn of Rosa roxburghii was observed by paraffin section. Leaves of Rosa roxburghii was used to synthesize cDNA based on the manufacturer's instructions of RACE. Subsequently RrGL2 was made relative informatics analysis and the gene expression level was eva-luated. The results were as follows:(1)The base cells continuously divided at the early stage of flower bud, then outward developed. The middle cells continued to become thinner and longer to form a “needle” structure. In the early stage of flower bud formation, the cells in the base of the thorn structure first divided continuously and continued to develop outwards. The cells in the middle became thinner and longer, forming a “needle” structure. The lignification gradually was observed at the top cells to make the prickles hard.(2)The full lengths of RrGL2 was 2 292 bp by RACE, encoding 763 amino acids.(3)The RrGL2 had a structure of Homeodomain and StAR phosphatidylcholine transfer protein, which is likely to regulate the development of Rosa roxburghii prickles. Then, a search for homologous species in the NCBI databases revealed a high similarity of amino acid homology encoded by the RrGL2 with other Rosa species, and phylogenic analysis revealed a close relationship of structure domains between Rosa roxburghii and Fragaria vesca.(4)Finally, real-time PCR analysis showed that the relative expression value of RrGL2 in fruit prickles after seven weeks after flowering was the highest, almost respectively 7.87 times and 2.10 times than that during three weeks and during five weeks after flowering. RrGL2, a prickles-forming gene acted to regulate the morphology and development of prickles. Therefore, the function of RrGL2 is closely related to thorn formation. These results could provide theoretical basis for thorn formation and development. |
| Key words: Rosa roxburghii, trichomes, prickles, RrGL2, gene expression |
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