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园艺学报 ›› 2013, Vol. 40 ›› Issue (1): 145-154.

• 研究报告 • 上一篇    下一篇

唐菖蒲质膜水孔蛋白基因GhPIP1;1 的克隆及表达分析

林燕飞, 李红梅, 丁岳练, 黄新敏, 冼锡金, 何生根   

  1. (仲恺农业工程学院生命科学学院,广州 510225)
  • 出版日期:2013-01-25 发布日期:2013-01-25

Cloning and Expression Analysis of a Plasma Membrane Aquaporin Gene GhPIP1;1 in Cut Gladiolus Flowers

 LIN  Yan-Fei, LI  Hong-Mei, DING  Yue-Lian, HUANG  Xin-Min, XIAN  Xi-Jin, HE  Sheng-Gen   

  1. (College of Life Sciences,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China)
  • Online:2013-01-25 Published:2013-01-25

摘要: 采用RT-PCR和RACE技术从唐菖蒲Gladiolus hybridus‘Eerde’)花瓣中克隆得到1个质膜内在蛋白(plasma membrane intrinsic proteins,PIPs)类的水孔蛋白基因,命名为GhPIP1;1。该基因cDNA 全长1 130 bp,包含867 bp完整开放阅读框(ORF),编码288个氨基酸。克隆和分析相应的gDNA序列(2 098 bp)表明,其包含由4个外显子和3个内含子组成的编码区序列。氨基酸序列分析表明GhPIP1;1具有水孔蛋白家族高度保守的2个NPA(Asn-Pro-Ala)基序。同源性分析显示GhPIP1;1氨基酸序列与同科的荷兰鸢尾(Iris hollandica)PIP1氨基酸序列的同源性达94%。半定量RT-PCR分析表明,GhPIP1;1在唐菖蒲花瓣、雄蕊、雌蕊、茎、苞片和叶片等均有表达,但表达量以花瓣中最高,叶片中最低;GhPIP1;1在花蕾至花朵盛开期间的表达水平较高且变化不明显,但在花朵盛开后的萎蔫过程中表达水平明显降低。

关键词: 唐菖蒲, 水孔蛋白, 分子特征, 基因表达

Abstract: A plasma membrane intrinsic proteins(PIPs)gene,designated GhPIP1;1,was cloned from petals on gladiolus(Gladiolus hybridus‘Eerde’)flowers by RT-PCR and RACE. The full cDNA sequence of GhPIP1;1 is 1 130 bp,containing an open reading frame(867 bp)and encoding a protein of 288 amino acids. The GhPIP1;1 genomic DNA(2 098 bp)was also cloned,which contains 4 exons and 3 introns in its coding sequence. Two highly conserved NPA(Asn-Pro-Ala)motifs of aquaporins were found in GhPIP1;1. Homology of amino acids sequences between GhPIP1;1 and PIP1 from Iris hollandica was up to 94%. The semi-quantitative RT-PCR analysis showed that GhPIP1;1 gene was expressed in petals,stamina,pistils,stems,bracts,and leaves on gladiolus flowering stems,and the expression level was the highest in petals,and the lowest in leaves. Then after further expression analysis of GhPIP1;1 gene in petals of gladiolus florets during opening process,it was found that it maintained high and relatively stable levels from bud stage to flower full opening,however it was obviously down-regulated during the subsequent flower wilting.

Key words: gladiolus, Gladiolus hybridus, aquaporin, molecular characterization, gene expression

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