Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, Wuhan 430079, China.
Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Sciences, Central China Normal University, Wuhan 430079, China.
Plant Physiol Biochem. 2014 Feb;75:70-9. doi: 10.1016/j.plaphy.2013.12.003. Epub 2013 Dec 17.
In this study, a cDNA, GhMADS14, encoding a typical MADS-box protein with 223 amino acids was isolated from a cotton cDNA library. Fluorescent microscopy indicated that the GhMADS14 protein was localized in the cell nucleus. GhMADS14 was specifically expressed in the elongating fibers, and its expression was gradually enhanced at early stages of fiber elongation and reached its peak in 9-10 DPA fibers. Overexpression of GhMADS14 in Arabidopsis hindered plant growth. Measurement and statistical analysis revealed that hypocotyl length of GhMADS14 transgenic seedlings was significantly reduced, and the height of the mature transgenic plants was remarkably less than that of the wild type. Furthermore, expression of GA 20-oxidase (AtGA20ox1 and AtGA20ox2) and GA 3-oxidase (AtGA3ox1 and AtGA3ox2) genes was remarkably reduced, whereas AtGA2ox1 and AtGA2ox8 were dramatically up-regulated in the transgenic plants, compared with the wild type. These results suggested that overexpression of GhMADS14 in Arabidopsis may alter expression levels of the genes related to GA biosynthetic and metabolic pathways, resulting in the reduction of endogenous GA amounts in cells. As a result, the transgenic plants grew slowly and display a GA-deficient phenotype.
在这项研究中,从棉花 cDNA 文库中分离出一个编码具有 223 个氨基酸的典型 MADS 盒蛋白的 cDNA,命名为 GhMADS14。荧光显微镜观察表明,GhMADS14 蛋白定位于细胞核中。GhMADS14 特异性表达在伸长纤维中,其表达在纤维伸长的早期阶段逐渐增强,在 9-10 DPA 纤维中达到峰值。GhMADS14 在拟南芥中的过表达会阻碍植物生长。测量和统计分析表明,GhMADS14 转基因幼苗的下胚轴长度显著缩短,成熟转基因植株的高度明显低于野生型。此外,与野生型相比,转基因植株中 GA 20-氧化酶(AtGA20ox1 和 AtGA20ox2)和 GA 3-氧化酶(AtGA3ox1 和 AtGA3ox2)基因的表达显著降低,而 AtGA2ox1 和 AtGA2ox8 则显著上调。这些结果表明,GhMADS14 在拟南芥中的过表达可能改变了与 GA 生物合成和代谢途径相关基因的表达水平,导致细胞内内源 GA 含量减少。因此,转基因植株生长缓慢,表现出 GA 缺陷表型。