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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。
One aquaporin gene of plasma membrane intrinsic proteins (PIPs) was cloned from Gladiolus hybridus’Eerde ’petals by RT-PCR and RACE techniques and named as GhPIP1. The full-length cDNA of this gene is 1 130 bp in length and contains 867 bp complete open reading frame (ORF) encoding 288 amino acids. Cloning and analysis of the corresponding gDNA sequence (2098 bp) indicated that it contains a coding sequence consisting of 4 exons and 3 introns. Amino acid sequence analysis revealed that GhPIP1; 1 has two NPA (Asn-Pro-Ala) motifs highly conserved among aquaporin families. Homology analysis showed that the amino acid sequence of GhPIP1; 1 shared 94% homology with the Iris hollandica PIP1 amino acid sequence in the same family. Semi-quantitative RT-PCR analysis showed that GhPIP1; 1 was expressed in the petals, stamens, pistils, stems, bracts and leaves of gladiolus, but its expression level was the highest among the petals and the lowest among the leaves. GhPIP1; During the bloom period, the expression level was higher and did not change significantly, but the expression level was significantly decreased during the wilting process after the flower was full bloom.