小麦胁迫相关蛋白基因TaSAP12-D的耐盐性分析

胁迫相关蛋白（SAPs）是一类具有A20/AN1锌指结构域的蛋白,在植物中主要参与逆境胁迫响应。为探究小麦胁迫相关蛋白基因TaSAP12-D在耐盐胁迫中的功能,本研究以小麦品种旱选10号为试材,克隆得到TaSAP12-D基因,利用农杆菌瞬时注射烟草叶片进行亚细胞定位,利用实时荧光定量PCR(qRT-PCR)进行不同组织和盐胁迫条件下的表达模式分析,利用蘸花法将TaSAP12-D转化到拟南芥（Arabidopsis thaliana L.）中并进行耐盐性分析。结果表明,TaSAP12-D基因全长519 bp,编码172个氨基酸,预测蛋白分子量为18.41 kDa,等电点为9.21。亚细胞定位显示,TaSAP12-D在烟草细胞核和细胞质中均有表达。qRT-PCR结果显示,TaSAP12-D在小麦萌发期和幼苗期的胚芽、根和叶中均有表达,其中在幼苗期的叶中表达量最高。在盐胁迫条件下,TaSAP12-D的表达量显著上调。在150 mmol·L-1NaCl处理条件下,过表达TaSAP12-D拟南芥的存活率显著提高,表明TaSAP12-D可以增强转基因拟南芥的耐盐性。另外,在转基因拟南芥中盐胁迫相关...

Identification and Analysis of Salt-tolerance of Stress Associated Protein Gene (TaSAP12-D) From Wheat

Stress associated proteins （SAPs） are a group of A20/AN1 zinc-finger domain-containing proteins which are mainly involved in response to abiotic stresses in plants. In order to explore the function of TaSAP12-D in response to salt stress, a stress associated protein gene designated TaSAP12-D was cloned from wheat （Trticum aestivum L.） cultivar Hanxuan 10. The TaSAP12-D：：GFP fusion protein construct was transferred into tobacco leavesthrough Agrobacterium tumefaciens mediated transfromation for subcellular localization. Quantitative real-time PCR was performed to analyze the expression patterns consisting of different tissues and leaves with NaCl treatment. To identify and analysis of salt-tolerance of TaSAP12-D, transgenic Arabidopsis plants were generated by floral infiltration. The results indicated that the full-length sequence of TaSAP12-D gene was 519 bp, encoding a 172-amino acid protein. The predicted relative molecular weight of the protein was 18.41 kDa and the isoelectric point was 9.21. Subcellular localization showed that TaSAP12-D was located in the nucleus and cytoplasm in tobacco leaf cells. The expression of TaSAP12-D was detected in different tissues, including plumule, root and leaf at the germination and seedling stages, and the highest expression occurred in the leaf tissues at the seedling stage. Furthermore, the transcript levels of TaSAP12-D were inducible by salt stress. Under treatment of 150 mmol·L^-1 NaCl, the survival rate of transgenic Arabidopsis plants was significantly higher than that of control. Overexpression of TaSAP12-D in Arabidopsis results in enhanced salt tolerance. In addition, the expression of salt stress-related genes （AtP5CS1, AtRD29A, AtLEA, AtSOS1, AtNHX1 and AtHKT） was significantly upregulated in transgenic Arabidopsis plants. It is speculated that TaSAP12-D can improve tolerance to salt stress by regulating the expression of salt stress-related genes. The results explored the molecular modulation mechanism of TaSAP12-D in response to salt stress, and provided candidate gene for improving salt tolerance in crops.