野生毛葡萄水通道蛋白基因VhPIPs的克隆与组织特异性表达研究

Cloning and Tissue Specificity Expression of the Aquaporin Genes from Wild Vitis quinquangularis

从野生毛葡萄组织中克隆了9个质膜水通道蛋白基因(VhPIP)的c DNA全长序列,并研究了其生物信息学特征及组织特异性表达。氨基酸序列分析表明,9个VhPIP分为PIP1和PIP2两类;均含有MIP蛋白家族保守的信号序列SGGHINPAVT和两个NPA(Asn-Pro-Ala)单元,均为稳定性蛋白、疏水性蛋白,且每个VhPIP均含有6个跨膜区。实时荧光定量RT-PCR分析表明,这9个VhPIP在抗旱的毛葡萄‘花溪–9’根、茎、叶中均有表达,其中VhPIP1;1、VhPIP1;2、VhPIP1;4、VhPIP2;1、VhPIP2;2和VhPIP2;4在其根中的表达量最多,可能与根部水分运输的调节密切相关;VhPIP1;5和VhPIP2;3在其叶中的表达量最多,可能主要参与叶片的水分运输,与叶片的蒸腾作用密切相关;VhPIP1;3在根、茎和叶中的表达量无显著性差异;相对于其他基因的表达,VhPIP1;1和VhPIP2;4在根中表达量最高,这两个基因可能在水分运输过程中起较大作用。

Nine plasma membrane aquaporin genes named VhPIPs were cloned from wild Vitis quinquangularis and their bioinformatics features and tissue specificity expressions were studied in order to study the structural features of aquaporin gene in wild grape and its role in the process of drought resistance. Amino acid sequence analysis showed that these 9 VhPIP genes can be divided into 2 groups，PIP1 and PIP2，signal sequence SGGHINPAVT contain MIP protein family conserved and 2 NPA（Asn-Pro-Ala）units，every protein is stable and hydrophobic，and each VhPIP contains 6 trans membrane domains. The real-time RT-PCR result showed they expressed in roots，stems and leaves. And VhPIP1;1，VhPIP1;2，VhPIP1;4，VhPIP2;1，VhPIP2;2 and VhPIP2;4 had higher expression level in the roots of ‘Huaxi-9’than other tissues，they may be closely related to the regulation of root water transportation. VhPIP1;5 and VhPIP2;3 had the highest expression level in leaves，these 2 genes may be mainly related to the water transportation and the transpiration of leaf. The expression level of VhPIP1;3 had no significance in roots，stems and leaves. Compared to the expression of other genes，VhPIP1;1 and VhPIP2;4 had the highest expression in roots，which indicated that they may play a greater role in the water transportation.