菊花CmPIN1同源基因的克隆与表达分析

发布时间：2018-06-24 04:52

  本文选题：菊花 + CmPIN　； 参考：《中国农业大学学报》2016年03期

【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 761bp,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5μmol/L的NAA 6h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。
[Abstract]:The full-length CmPIN1 gene of Chrysanthemum morifolium Jinba' was obtained by race and homologous cloning. The expression of CmPIN1 in different tissues and organs was compared by qRT-PCR, and the response of CmPIN1 to NAA and topping was studied. The results showed that the ORF of the CmPIN1 gene was 1761 BP, encoding 586 amino acids, and the transmembrane domain was located at the N-terminal and C-terminal of the protein, and the similarity between CmPIN1 protein and ZvPIN1 protein of Chrysanthemum chrysanthemum was the highest. The results of subcellular localization showed that CmPIN1 was located on cell membrane 2) and the expression of CmPIN1 was relatively high in stems and buds of chrysanthemum, meanwhile, CmPIN1 was induced by auxin in stem segments in vitro, and the expression of CmPIN1 was decreased after topping. CmPIN1 gene expression in the first lateral bud of the near apical bud recovered preferentially and tended to replace the apical bud as a new auxin source after the ceilings were removed by 5 渭 mol / L NAA for 6 h after the restoration of CmPIN1 expression in the chrysanthemum plants, and the results showed that the expression of CmPIN1 gene in the first lateral bud of the near apical bud recovered preferentially. In order to maintain the polar transport of auxin in the main stem, when the auxin transport channel in the main stem was opened again, the expression of CmPIN1 was gradually restored in the stem nodes, and the expression of CmPIN1 was slowly restored in the first internode near the top of the stem. CmPIN1 was involved in the polar transport of auxin in the main stem of chrysanthemum and the maintenance of apical dominance, which indirectly inhibited the elongation of lateral buds.
【作者单位】： 中国农业大学农学与生物技术学院;
【基金】：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)
【分类号】：S682.11
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本文编号：2060101