Chen Jingguang, Zhang Yong, Tan Yawen, Zhang Min, Zhu Longlong, Xu Guohua, Fan Xiaorong
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.
Key Laboratory of Plant Nutrition and Fertilization in Low-Middle Reaches of the Yangtze River, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.
Plant Biotechnol J. 2016 Aug;14(8):1705-15. doi: 10.1111/pbi.12531. Epub 2016 Jan 30.
The importance of the nitrate (NO3-) transporter for yield and nitrogen-use efficiency (NUE) in rice was previously demonstrated using map-based cloning. In this study, we enhanced the expression of the OsNRT2.1 gene, which encodes a high-affinity NO3- transporter, using a ubiquitin (Ubi) promoter and the NO3--inducible promoter of the OsNAR2.1 gene to drive OsNRT2.1 expression in transgenic rice plants. Transgenic lines expressing pUbi:OsNRT2.1 or pOsNAR2.1:OsNRT2.1 constructs exhibited the increased total biomass including yields of approximately 21% and 38% compared with wild-type (WT) plants. The agricultural NUE (ANUE) of the pUbi:OsNRT2.1 lines decreased to 83% of that of the WT plants, while the ANUE of the pOsNAR2.1:OsNRT2.1 lines increased to 128% of that of the WT plants. The dry matter transfer into grain decreased by 68% in the pUbi:OsNRT2.1 lines and increased by 46% in the pOsNAR2.1:OsNRT2.1 lines relative to the WT. The expression of OsNRT2.1 in shoot and grain showed that Ubi enhanced OsNRT2.1 expression by 7.5-fold averagely and OsNAR2.1 promoters increased by about 80% higher than the WT. Interestingly, we found that the OsNAR2.1 was expressed higher in all the organs of pUbi:OsNRT2.1 lines; however, for pOsNAR2.1:OsNRT2.1 lines, OsNAR2.1 expression was only increased in root, leaf sheaths and internodes. We show that increased expression of OsNRT2.1, especially driven by OsNAR2.1 promoter, can improve the yield and NUE in rice.
先前通过图位克隆证明了硝酸盐(NO3-)转运蛋白对水稻产量和氮利用效率(NUE)的重要性。在本研究中,我们使用泛素(Ubi)启动子和OsNAR2.1基因的NO3-诱导型启动子来驱动OsNRT2.1基因在转基因水稻植株中的表达,该基因编码一种高亲和力的NO3-转运蛋白。与野生型(WT)植株相比,表达pUbi:OsNRT2.1或pOsNAR2.1:OsNRT2.1构建体的转基因株系的总生物量增加,产量分别提高了约21%和38%。pUbi:OsNRT2.1株系的农业氮利用效率(ANUE)降至野生型植株的83%,而pOsNAR2.1:OsNRT2.1株系的ANUE提高到野生型植株的128%。相对于野生型,pUbi:OsNRT2.1株系向籽粒中的干物质转运减少了68%,而pOsNAR2.1:OsNRT2.1株系增加了46%。OsNRT2.1在地上部和籽粒中的表达表明,Ubi平均使OsNRT2.1表达提高了7.5倍,OsNAR2.1启动子使其表达比野生型高出约80%。有趣的是,我们发现OsNAR2.1在pUbi:OsNRT2.1株系的所有器官中表达更高;然而,对于pOsNAR2.1:OsNRT2.1株系,OsNAR2.1表达仅在根、叶鞘和节间增加。我们表明,OsNRT2.1表达的增加,特别是由OsNAR2.1启动子驱动的,可提高水稻的产量和氮利用效率。