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作物学报 ›› 2015, Vol. 41 ›› Issue (05): 673-682.doi: 10.3724/SP.J.1006.2015.00673

• 作物遗传育种·种质资源·分子遗传学 •    下一篇

小麦脂质转运蛋白基因TaLTP克隆及功能分析

李倩1,2,王景一2,毛新国2,李昂2,高丽锋2,刘惠民1,景蕊莲2,*   

  1. 1山西大学生物工程学院, 山西太原 030006; 2农作物基因资源与基因改良国家重大科学工程 / 中国农业科学院作物科学研究所, 北京 100081
  • 收稿日期:2015-01-02 修回日期:2015-03-19 出版日期:2015-05-12 网络出版日期:2015-03-30
  • 通讯作者: 景蕊莲, E-mail: jingruilian@caas.cn, Tel: 010-82105829
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2011AA100501, 2012AA10A308)资助。

Cloning and Functional Analysis of Lipid Transfer Protein Gene TaLTP in Wheat

LI Qian1,2,WANG Jing-Yi2,MAO Xin-Guo2,Li Ang2,GAO Li-Feng2,LIU Hui-Min1,JING Rui-Lian2,*   

  1. 1 College of Bioengineering, Shanxi University, Taiyuan 030006, China; 2 National Key Facility for Crop Gene Resources and Genetic Improvement / Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing
  • Received:2015-01-02 Revised:2015-03-19 Published:2015-05-12 Published online:2015-03-30
  • Contact: 景蕊莲, E-mail: jingruilian@caas.cn, Tel: 010-82105829

摘要:

脂质转运蛋白(lipid transfer protein, LTP)是广泛存在于植物中的一类小分子蛋白,因其能在植物细胞膜间运输脂类物质而得名,在植物角质层合成和适应多种环境胁迫中起重要作用。本研究利用同源克隆方法得到TaLTP-scDNA全长序列510 bp,开放阅读框为339 bp,编码112个氨基酸。利用RIL群体将TaLTP-s定位在染色体1A上,距离两侧标记WMC449WMC93的遗传距离分别为2.1 cM5.9 cM。通过原生质体亚细胞定位显示TaLTP-s定位于细胞膜和细胞质中。小麦开花期TaLTP-s在小花中表达量较高,尤其是在花药中的表达量远高于在根和叶中。在ABAPEGNaCl4低温胁迫诱导下,小麦TaLTP-s均上调表达,可能与抗逆性调节相关。在高盐胁迫处理下,转基因拟南芥幼苗的细胞膜稳定性和存活率显著高于野生型。研究结果为利用小麦脂质转运蛋白基因改良作物抗逆性奠定了基础。

关键词: 小麦, 脂质转运蛋白, 基因克隆, 非生物胁迫, 耐盐性

Abstract:

Lipid transfer protein (LTP) is a kind of small molecular protein, which name stands for its ability to transfer lipid between cell membranes in plant. LTP plays a key role in cuticle synthesis and adaptation to abiotic stress. We cloned a full-length cDNA sequence of TaLTP gene encoding a lipid transfer protein from wheat (Triticum aestivum L.), which length is 510 bp with a 339 bp open reading frame. The cDNA sequence contains 112 amino acids with a N-terminal signal peptide within the first 25 amino acids. This gene contains eight cysteine residues conserved in amino acid sequences. TaLTP-s was detected to be located between markers WMC449 and WMC93 on chromosome 1A in wheat, with genetic distances of 2.1 cM and 5.9 cM, respectively. The result of subcellular localization exhibited that TaLTP-s is located in the cell membrane and cytoplasm. TaLTP-s was expressed in all tissues at flowering stage, including leaf, root, floret, anther and pistil, and mature seeds of wheat. The highest expression level was identified in the floret, especially in the anther. TaLTP-s was up-regulated by ABA, PEG, NaCl and 4ºC treatments which indicates that TaLTP-s is involved in different signal pathways responding to abiotic stress. Arabidopsis thaliana overexpressing TaLTP-s showed higher cell membrane stability and survival rate than the controls under salinity stress. These results provide a candidate gene for wheat improvement in response to salt and other abiotic stresses.

Key words: Wheat, TaLTP-s, Gene cloning, Abiotic stress, Salt tolerance

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