玉米抗逆基因ZmqLTG3-1的克隆及功能分析

doi:10.16035/j.issn.1001-7283.2016.04.008

摘要/Abstract

摘要：

以玉米自交系豫82为材料,同源克隆了与水稻qLTG3-1基因同源的玉米ZmqLTG3-1基因。序列分析发现,该基因cDNA序列全长606bp,开放阅读框441bp,编码包含147个氨基酸的蛋白。该蛋白含有AAI_LTSS超蛋白家族的LTP（含8个半胱氨酸）保守结构,分子量为14.16kDa,等电点是8.3,与高粱、水稻的相似度分别为92%和88%。荧光定量RT-PCR分析结果表明,ZmqLTG3-1在玉米胚中的表达量最高,其次是茎尖,在叶、根等器官中的表达量较低;随着胚萌发时间的延长,其表达量逐渐增加,在萌发24h时呈现最高丰度表达,之后表达丰度与萌发时间呈现规律性下降。亚细胞定位结果表明,该基因所编码的产物被定位在细胞膜上,是一跨膜蛋白。通过农杆菌介导的花序侵染法将过表达载体OE-ZmqLTG3-1和空载体pCAMBIA1304转入拟南芥,在诸如干旱、高温和盐等非生物胁迫下,与对照相比,T₃转基因植株的成活率显著提高,说明ZmqLTG3-1基因对于抵抗非生物胁迫发挥了重要作用。

关键词: 玉米, ZmqLTG3-1, 克隆, 功能分析

Abstract:

By the method of orthologus gene cloning,the ZmqLTG3-1 gene in maize was successfully cloned, which was homologous to the qLTG3-1 gene in rice. The sequence analysis indicated that the full-length cDNA was composed of 606bp, including an open reading frame (ORF) of 441bp, which encoded 147 amino acids. The protein contained LTP conservative structure of the extra protein family AAI-LTSS, and protein molecular weight and isoelectric point were 14.16kDa and 8.3, respectively, with the similarity of amino acid sequence for ZmqLTG3-1 with sorghum and rice being 92% and 88%. The results of quantitative RT-PCR showed that the highest expression level was in the embryo, then in the stem tip, lower was in the leaf and root. The highest expression abundance at 24h and then regular down along with germination time. The result of subcell location revealed that the product of ZmqLTG3-1 gene encoding is positioned on the cell membrane and it was a trans-membrane protein. To carry the preliminary validation of its function,transgenic T₃ generation, trans-empty vector and wild type Arabidopsis thaliana as materials were studied. Results showed that in different abiotic conditions, such as drought, high temperature and salt, compared with control group, the performance and the survival rate of transgenic plant are significantly higher. All these suggest that ZmqLTG3-1 gene plays an important role in resisting abiotic stress.

Key words: Maize (Zea mays L.), ZmqLTG3-1, Cloning, Function analysis

韩赞平,陈彦惠,郭书磊,祖小峰,王顺喜,赵西拥. 玉米抗逆基因ZmqLTG3-1的克隆及功能分析[J]. 作物杂志, 2016 (4): 47–55

Zanping Han,Yanhui Chen,Shulei Guo,Xiaofeng Zu,Shunxi Wang,Xiyong Zhao. Cloning and Function Analysis of Resistance Gene ZmqLTG3-1 in Maize[J]. Crops, 2016(4): 47–55

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