National Key Laboratory of Crop Genetic Improvement, and Microelement Research Center, Huazhong Agricultural University, Wuhan, China.
PLoS One. 2013;8(4):e60801. doi: 10.1371/journal.pone.0060801. Epub 2013 Apr 3.
Phytate is the major storage form of organic phosphorus in soils and plant seeds, and phosphorus (P) in this form is unavailable to plants or monogastric animals. In the present study, the phytase genes phyA and appA were introduced into Brassica napus cv Westar with a signal peptide sequence and CaMV 35S promoter, respectively. Three independent transgenic lines, P3 and P11 from phyA and a18 from appA, were selected. The three transgenic lines exhibited significantly higher exuded phytase activity when compared to wild-type (WT) controls. A quartz sand culture experiment demonstrated that transgenic Brassica napus had significantly improved P uptake and plant biomass. A soil culture experiment revealed that seed yields of transgenic lines P11 and a18 increased by 20.9% and 59.9%, respectively, when compared to WT. When phytate was used as the sole P source, P accumulation in seeds increased by 20.6% and 46.9% with respect to WT in P11 and a18, respectively. The P3 line accumulated markedly more P in seeds than WT, while no significant difference was observed in seed yields when phytate was used as the sole P source. Phytase activities in transgenic canola seeds ranged from 1,138 to 1,605 U kg(-1) seeds, while no phytase activity was detected in WT seeds. Moreover, phytic acid content in P11 and a18 seeds was significantly lower than in WT. These results introduce an opportunity for improvement of soil and seed phytate-P bioavailability through genetic manipulation of oilseed rape, thereby increasing plant production and P nutrition for monogastric animals.
植酸是土壤和植物种子中有机磷的主要储存形式,这种形式的磷对植物或单胃动物是不可用的。在本研究中,分别使用带有信号肽序列和 CaMV 35S 启动子的植酸酶基因 phyA 和 appA 导入甘蓝型油菜品种 Westar。选择了三个独立的转基因株系,phyA 的 P3 和 P11 以及 appA 的 a18。与野生型(WT)对照相比,这三个转基因株系表现出明显更高的分泌植酸酶活性。石英砂培养实验表明,转基因油菜具有显著提高的磷吸收和植物生物量。土壤培养实验表明,与 WT 相比,转基因株系 P11 和 a18 的种子产量分别增加了 20.9%和 59.9%。当植酸作为唯一的磷源时,与 WT 相比,P11 和 a18 种子中的磷积累分别增加了 20.6%和 46.9%。与 WT 相比,P3 株系在种子中积累的磷明显更多,但当植酸作为唯一的磷源时,种子产量没有明显差异。转基因油菜种子中的植酸酶活性范围为 1138-1605 U kg(-1)种子,而 WT 种子中未检测到植酸酶活性。此外,P11 和 a18 种子中的植酸含量明显低于 WT。这些结果为通过遗传操作油菜来提高土壤和种子植酸-P 的生物有效性提供了机会,从而提高了植物生产和单胃动物的磷营养。