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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。
Chrysanthemum morifolium’Jinba ’was used as a material to obtain the full length of CmPIN1 gene by RACE technique and homologous cloning method. The expression of CmPIN1 in different tissues and organs was compared by qRT-PCR and the response to CmPIN1 Apply NAA and go top study. The results showed that: 1) The ORF of CmPIN1 gene was 1 761 bp in length and encoded a protein of 586 amino acids. The transmembrane domain was located at the N- and C-terminus of the protein. The CmPIN1 protein had the highest similarity with ZvPIN1 protein The results of cell localization showed that CmPIN1 was located on the cell membrane. 2) The expression of CmPIN1 showed that the gene had a relatively high expression level in stem and bud of chrysanthemum, while the CmPIN1 in in vitro stem was induced by auxin; While the expression of CmPIN1 was restored after applying 5μmol / L NAA for 6h. 3) The de-toping test of chrysanthemum plants showed that the expression of CmPIN1 gene in the first lateral bud near the tip preferentially recovered, In the process of replacing the terminal bud, a new auxin source was added 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 level of CmPIN1 gradually recovered in the stem, The expression of CmPIN1 in one internode recovered more slowly. CmPIN1 can be seen to participate in the polar transport of auxin in the main chrysanthemum stem, as well as the maintenance of the top advantage, indirect inhibition of lateral bud elongation.