| 摘要: |
| 以切花菊'神马'( Chrysanthemum morifolium 'Jinba')为材料,利用RACE技术与同源克隆方法获得菊花 CmPIN1 基因全长,通过qRT-PCR技术比较 CmPIN1 在不同组织器官的表达,同时对 CmPIN1 响应外施NAA和去顶进行研究。结果表明:1) CmPIN1 基因ORF全长1 761 bp,编码586个氨基酸,跨膜域位于蛋白N端与C端;通过蛋白序列比对分析,菊花CmPIN1蛋白与百日草ZvPIN1蛋白相似性最高;亚细胞定位结果显示CmPIN1位于细胞膜;2)对 CmPIN1 表达分析表明,该基因在菊花茎部与芽部具有相对较高的表达量,同时,在离体茎段中的 CmPIN1 受到生长素诱导;去顶后,CmPIN1表达量降低,而施加5 μmol/L的 NAA 6 h后 CmPIN1 表达恢复;3)对菊花植株进行去顶试验表明,去顶后,近顶端第一个侧芽内 CmPIN1 基因表达量优先恢复,趋向于代替顶芽成为新的生长素源,以维持主茎中生长素极性运输;当主茎中的生长素运输通道再次打开,茎节中 CmPIN1 的表达量均逐渐恢复,其中,在近顶端第一个节间中 CmPIN1 的表达量恢复较缓慢。可见 CmPIN1 参与菊花主茎中生长素的极性运输,以及顶端优势的维持,间接抑制侧芽伸长。 |
| 关键词: 菊花 CmPIN1 基因表达 生长素 去顶 |
| DOI：10.11841/j.issn.1007-4333.2016.03.09 |
| 投稿时间：2015-03-19 |
| 基金项目:国家'863'计划项目(2011AA100208); 农业部"引进国际先进农业科学技术"重点项目(2008-G3) |
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| Isolation and expression analysis of auxin efflux transporter CmPIN1 in Chrysanthemum (Chrysanthemum morifolium 'Jinba') |
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LV Su-hui, XI Lin, NIE Jing, WEN Chao, YUAN Cun-quan, MA Nan, ZHAO Liang-jun
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| (College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China) |
| Abstract: |
| Using Rapid Amplification of cDNA Ends (RACE) approach, the full-length cDNA sequences of an auxin efflux transporter gene was isolated from Chrysanthemum (Chrysanthemum morifolium 'Jinba') and named as CmPIN1 .To characterize CmPIN1, we tested spatial-temporal expression pattern of CmPIN1 in chrysanthemum, as well as effects of auxin and decapitation treatments on CmPIN1 expression using qRT-PCR.The Open Reading Frame of CmPIN1 was 1 761 bp in length, encoding 586 amino acids.The predicted transmembrane domains located in both the C-terminal and N-terminal.Multiple sequence alignment and phylogenetic analysis showed that putative CmPIN1 protein shared the highest similarity with PIN1 protein from Zinnia violacea.Subcellular localization analysis showed that CmPIN1 located in membrane system.Expression of CmPIN1 was higher in stems and buds than in other organs, and it was induced by auxin.After decapitation, CmPIN1 level dropped in stems, while increased after 5 μmol/L NAA was applied to the decapitated plants for 6 hours. CmPIN1 expression level in the axillary bud, which was close to the apical bud, recovered firstly, implying that the axillary bud was tending to become new auxin source and function as apical bud.Once the auxin gradient re-established, the expression level of CmPIN1 was recovered gradually, whereas it recovered more slowly in the first internodes close to the apical bud.Taken together, we concluded that CmPIN1 was mainly expressed in stem, and involved in auxin polar transportation and maintaining of apical dominance to inhibit axillary bud outgrowth indirectly. |
| Key words: chrysanthemum CmPIN1 gene expression auxin decapitation |
