唐菖蒲质膜水孔蛋白基因GhPIP1;1的克隆及表达分析  

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

作　　者：林燕飞[1] 李红梅[1] 丁岳练[1] 黄新敏[1] 冼锡金[1] 何生根[1] 

机构地区：[1]仲恺农业工程学院生命科学学院,广州510225

出　　处：《园艺学报》2013年第1期145-154,共10页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31071829;31272193);广东省自然科学基金项目(10151022501000035;S2012010010418)

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。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.

关 键 词：唐菖蒲 水孔蛋白 分子特征 基因表达 

分 类 号：S681[农业科学—观赏园艺]