《植物生理学报》 2015, 51(9): 1405-1413

‘富士’苹果Ca²⁺/H⁺反向转运蛋白基因生物信息学及表达模式分析

程玉豆^1,2, 冯云霄^1,2, 关军锋^1,2,*

¹河北省农林科学院遗传生理研究所, 石家庄050051; ²河北省植物转基因研究中心, 石家庄050051

通信作者：关军锋；E-mail: junfeng-guan@263.net；Tel: 0311-87652132

摘　要：

本研究借助生物信息学分析从苹果基因组数据库中获得了6个含全长编码序列的Ca²⁺/H⁺反向转运蛋白(Ca²⁺/H⁺ antiporter, CAX)基因, 分别命名为MdCAX1、MdCAX2、MdCAX3、MdCAX4、MdCAX5和MdCAX6。基因和蛋白结构特性分析结果显示: 6个CAX在基因结构和蛋白特性等方面存在差异; MdCAXs隶属于I-A型和I-B 2个亚型, 均含有N-端自我抑制区(N-terminal regulatory region, NRR)、CaD功能域、C-端功能域、C功能域和D功能域。荧光定量PCR分析表明, MdCAX1在‘富士’苹果叶片中表达量表现为基部叶片>中部叶片>顶端叶片, 在果皮和果肉中的表达量低于叶片; MdCAX2在果皮和果肉中表达量高于叶片; MdCAX3在叶片和果实中的表达模式与MdCAX1相似, 但在叶片中的表达量低于MdCAX1; MdCAX4在各被检测样本中表达量无明显变化; MdCAX5和MdCAX6在果皮和果肉组织中的表达量随发育进程呈逐渐升高趋势。叶片中Ca²⁺含量在不同叶位叶片中差异显著, 果皮和果肉中Ca²⁺含量随着果实发育均呈下降趋势。上述研究结果表明: 6个MdCAXs基因在基因结构、蛋白特性以及表达模式上存在差异, MdCAX1和MdCAX3基因表达与‘富士’苹果叶片和果实Ca²⁺含量变化密切相关。

关键词：苹果; Ca²⁺/H⁺反向转运蛋白; 生物信息学分析; 钙; 基因表达

收稿：2015-03-09   修定：2015-04-24

资助：河北省科技支撑计划项目(14826820D)和农业科技创新人才队伍建设(2013055003)。

Bioinformatics and Expression Analysis of Ca²⁺/H⁺ Antiporter Gene Family in ‘Fuji’ Apple

CHENG Yu-Dou^1,2, FENG Yun-Xiao^1,2, GUAN Jun-Feng^1,2,*

¹Institute of Genetics and Physiology, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050051, China; ²Plant Genetic Engineering Center of Hebei Province, Shijiazhuang 050051, China

Corresponding author: GUAN Jun-Feng; E-mail: junfeng-guan@263.net; Tel: 0311-87652132

Abstract:

Six Ca²⁺/H⁺ antiporter (CAX) genes which contained full-length coding sequences were obtained from the apple genome database by bioinformatics analysis in this work, and designated as MdCAX1, MdCAX2, MdCAX3, MdCAX4, MdCAX5 and MdCAX6 respectively. The studies of gene and protein characteristics indicated that the gene formations and protein characteristics of the six MdCAXs were different. The MdCAXs were clustered into two groups: I-A type and I-B type, and all of the 6 CAXs contained N-terminal regulatory region (NRR), CaD domain, C-terminal domain, C domain and D domain. The results of real-time PCR showed that the expression level of MdCAX1 presented base leaf > middle leaf > top leaf, and showed lower levels in peel and flesh than that in leaves. The expression levels of MdCAX2 in peel and flesh were higher than that in leaf. The expression pattern of MdCAX3 was similar to MdCAX1 in leaf, but the expression level was lower than MdCAX1. No markedly change of MdCAX4 expression level was detected in all the tested simples. The expression levels of MdCAX5 and MdCAX6 gradually increased during the development of peel and flesh. The contents of Ca²⁺ increased gradually accompanied by the grade of maturity rising in leaf, while the Ca²⁺ content in peel and flesh showed a downtrend in fruit during the course of development. The above-mentioned results showed that the gene formations, protein characteristics and expression patterns of the six MdCAXs were different, and the expression levels of MdCAX1 and MdCAX3 were closely related to Ca²⁺ content in leaf of ‘Fuji’ apple.

Key words: apple; Ca²⁺/H⁺ antiporter; bioinformatics analysis; calcium; gene expression