当前位置: 首页 > 文章 > 唐菖蒲质膜水孔蛋白基因GhPIP1;1的克隆及表达分析 园艺学报 2013,40 (1) 145-154
Position: Home > Articles > Cloning and Expression Analysis of a Plasma Membrane Aquaporin Gene GhPIP1;1 in Cut Gladiolus Flowers Acta Horticulturae Sinica 2013,40 (1) 145-154

唐菖蒲质膜水孔蛋白基因GhPIP1;1的克隆及表达分析

作  者:
林燕飞;李红梅;丁岳练;黄新敏;冼锡金;何生根
单  位:
仲恺农业工程学院生命科学学院
关键词:
唐菖蒲;水孔蛋白;分子特征;基因表达
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。
译  名:
Cloning and Expression Analysis of a Plasma Membrane Aquaporin Gene GhPIP1;1 in Cut Gladiolus Flowers
作  者:
LIN Yan-fei,LI Hong-mei * ,DING Yue-lian,HUANG Xin-min,XIAN Xi-jin,and HE Sheng-gen * (College of Life Sciences,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China)
关键词:
gladiolus;Gladiolus hybridus;aquaporin;molecular characterization;gene expression
摘  要:
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 stablelevels from bud stage to flower full opening,however it was obviously down-regulated during the subsequent flower wilting.

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