Wang Cuiting, Yang Yang, Wang Haihai, Ran Xiaojuan, Li Bei, Zhang Jiantao, Zhang Hongxia
National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, China.
Plant Biotechnol J. 2016 Sep;14(9):1838-51. doi: 10.1111/pbi.12544. Epub 2016 Mar 11.
In Arabidopsis thaliana and Oryza sativa, the cytochrome P450 (CYP) 714 protein family represents a unique group of CYP monooxygenase, which functions as a shoot-specific regulator in plant development through gibberellin deactivation. Here, we report the functional characterizations of PtCYP714A3, an OsCYP714D1/Eui homologue from Populus trichocarpa. PtCYP714A3 was ubiquitously expressed with the highest transcript level in cambium-phloem tissues, and was greatly induced by salt and osmotic stress in poplar. Subcellular localization analyses indicated that PtCYP714A3-YFP fusion protein was targeted to endoplasmic reticulum (ER). Expression of PtCYP714A3 in the rice eui mutant could rescue its excessive-shoot-growth phenotype. Ectopic expression of PtCYP714A3 in rice led to semi-dwarfed phenotype with promoted tillering and reduced seed size. Transgenic lines which showed significant expression of PtCYP714A3 also accumulated lower GA level than did the wild-type (WT) plants. The expression of some GA biosynthesis genes was significantly suppressed in these transgenic plants. Furthermore, transgenic rice plants exhibited enhanced tolerance to salt and maintained more Na(+) in both shoot and root tissues under salinity stress. All these results not only suggest a crucial role of PtCYP714A3 in shoot responses to salt toxicity in rice, but also provide a molecular basis for genetic engineering of salt-tolerant crops.
在拟南芥和水稻中,细胞色素P450(CYP)714蛋白家族代表了一组独特的CYP单加氧酶,其通过赤霉素失活在植物发育中作为茎特异性调节剂发挥作用。在此,我们报道了来自毛果杨的OsCYP714D1/Eui同源物PtCYP714A3的功能特性。PtCYP714A3在形成层-韧皮部组织中普遍表达,转录水平最高,并且在杨树中受到盐和渗透胁迫的强烈诱导。亚细胞定位分析表明,PtCYP714A3-YFP融合蛋白定位于内质网(ER)。PtCYP714A3在水稻eui突变体中的表达可以挽救其过度生长的茎表型。PtCYP714A3在水稻中的异位表达导致半矮化表型,分蘖增加,种子大小减小。显著表达PtCYP714A3的转基因系也比野生型(WT)植物积累更低水平的赤霉素。在这些转基因植物中,一些赤霉素生物合成基因的表达受到显著抑制。此外,转基因水稻植株表现出对盐的耐受性增强,并且在盐胁迫下地上部和根部组织中保留更多的Na(+)。所有这些结果不仅表明PtCYP714A3在水稻地上部对盐毒性反应中的关键作用,而且为耐盐作物的基因工程提供了分子基础。