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伸长棉纤维中质膜和液泡膜水通道蛋白的表征与表达

Characterization and expression of plasma and tonoplast membrane aquaporins in elongating cotton fibers.

作者信息

Liu Diqiu, Tu Lili, Wang Li, Li Yunjing, Zhu Longfu, Zhang Xianlong

机构信息

National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.

出版信息

Plant Cell Rep. 2008 Aug;27(8):1385-94. doi: 10.1007/s00299-008-0545-6. Epub 2008 Apr 5.

Abstract

Cotton fiber (Gossypium hirsutum L. and G. barbadense L.) is a good model for studies of plant cell elongation and cell wall biogenesis. Aquaporins are ancient membrane channel proteins that facilitate the permeation of water across biological membranes. We studied GhPIP1-2, encoding plasma membrane intrinsic protein, and GhgammaTIP1, encoding tonoplast intrinsic protein, during cotton fiber development. The full-length cDNAs of GhPIP1-2 and GhgammaTIP1 were obtained through 5' RACE. The deduced amino acid sequences of GhPIP1-2 and GhgammaTIP1 share high sequence identity with aquaporins from diverse plant species. Phylogenetic analysis of GhPIP1-2 and GhgammaTIP1 with other plant aquaporins showed that GhPIP1-2 belongs to the PIP1 group of the PIP subfamily and GhgammaTIP1 belongs to the gammaTIP group of the TIP subfamily. GhPIP1-2 and GhgammaTIP1 contain three and two introns, respectively. Genomic Southern blot analysis indicated that GhPIP1-2 and GhgammaTIP1 have several copies and multiple homologous genes in allotetraploid cotton. Northern blot analysis with gene-specific probes and real-time PCR demonstrated that GhPIP1-2 and GhgammaTIP1 are predominantly expressed during cotton fiber elongation, with the highest expression levels at 5 days post-anthesis. Moreover, expression patterns of the two genes in G. hirsutum and G. barbadense are similar, whereas the expression levels in G. barbadense are much lower than that in G. hirsutum. The high and preferential expression of GhPIP1-2 and GhgammaTIP1 during fiber cell elongation suggests that they may play important roles in supporting the rapid influx of water into vacuoles during cotton fiber cell expansion.

摘要

棉纤维(陆地棉和海岛棉)是研究植物细胞伸长和细胞壁生物合成的良好模型。水通道蛋白是古老的膜通道蛋白,可促进水穿过生物膜。我们研究了在棉纤维发育过程中编码质膜内在蛋白的GhPIP1-2和编码液泡膜内在蛋白的GhgammaTIP1。通过5' RACE获得了GhPIP1-2和GhgammaTIP1的全长cDNA。GhPIP1-2和GhgammaTIP1推导的氨基酸序列与来自不同植物物种的水通道蛋白具有高度的序列同一性。对GhPIP1-2和GhgammaTIP1与其他植物水通道蛋白的系统发育分析表明,GhPIP1-2属于PIP亚家族的PIP1组,GhgammaTIP1属于TIP亚家族的gammaTIP组。GhPIP1-2和GhgammaTIP1分别包含三个和两个内含子。基因组Southern杂交分析表明,GhPIP1-2和GhgammaTIP1在异源四倍体棉花中有多个拷贝和多个同源基因。用基因特异性探针进行的Northern杂交分析和实时PCR表明,GhPIP1-2和GhgammaTIP1在棉纤维伸长过程中主要表达,在开花后5天表达水平最高。此外,这两个基因在陆地棉和海岛棉中的表达模式相似,而在海岛棉中的表达水平远低于陆地棉。GhPIP1-2和GhgammaTIP1在纤维细胞伸长过程中的高表达和优先表达表明,它们可能在支持棉纤维细胞扩张过程中液泡快速吸水方面发挥重要作用。

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