作物学报2016，Vol.42Issue(9)：1282-1290,9.DOI:10.3724/SP.J.1006.2016.01282
小麦蛋白磷酸酶2A基因TaPP2AbB″-α启动子的克隆及表达分析
Cloning and Expression Analysis of Protein Phosphatase 2A GeneTaPP2AbB″-α Promoterin Wheat
摘要
Abstract
Protein phosphatase 2A (PP2A) is a heterotrimeric protein, consisting of a scaffolding subunit (A), a catalytic subunit (C), and a member of four families of regulatory subunits (B). PP2A plays significant roles in the pathway responding to abiotic stresses in plants.TaPP2AbB″-α, a member of regulatory subunit B″ in wheat (Triticum aestivum L.), enhanced root development and could develop more lateral roots in the gene overexpressedArabidopsis, especially under the osmotic stresses of mannitol and NaCl. In order to elucidate transcriptional regulatory mechanism of the promoter PB″α ofTaPP2AbB″-α, we isolated an 1899 bp full-length sequence of promoter PB″α from a drought-tolerant wheat cultivar Hanxuan 10. The PB″α sequence contained TATA-box, CAAT-box and a series ofcis-acting elements responding to drought and osmotic stresses, such as elements of EECCRCAH1 (from-1058 to-1052 bp), GCCCORE (from-1073 to-1068 bp) and MYCCONSE (from-1179 to-1174 bp). The full-length promoter PB″α and five 5′-end truncated PB″α promoters in different lengths fused with the reporter geneβ-glucuronidas (GUS) were transformed intoArabidopsis, respectively. The histochemical staining results showed that the full-length promoter PB″α, and the deletion fragments of PB″α-1545 and PB″α-1389 could driveGUS gene expression in shoots and roots of transgenicArabidopsis seedlings. As the result of quantitative fluorometric GUS assay, only the expression of PB″α, PB″α-1545 and PB″α-1389 could be up-regulated by salt and osmotic stresses in transgenicArabidopsis lines, and the active re-gion of PB″α promoter was located in the interval between-1389 bp and-946 bp. In conclusion, PB″α has strong basic promoter activity which is up-regulated significantly by salt and osmotic stresses. These findings contribute to the selection of a suitable promoter for crop improvement.关键词
小麦/蛋白磷酸酶2A/启动子/顺式作用元件/GUS组织化学染色/GUS定量分析Key words
Wheat/PP2A/Promoter/cis-acting Element/GUS histochemical staining/Quantitative fluorometric GUS assay引用本文复制引用
扆珩,李昂,刘惠民,景蕊莲..小麦蛋白磷酸酶2A基因TaPP2AbB″-α启动子的克隆及表达分析[J].作物学报,2016,42(9):1282-1290,9.基金项目
本研究由国家自然科学基金项目(31201206)和中国农业科学院创新工程项目资助。This study was supported by the National Natural Science Foundation of China (31201206) and the Agricultural Science and Technology Innovation Program (CAAS) （31201206）
