分类： 生物学 >> 植物学 >> 植物生物化学、植物生物物理学 提交时间： 2016-05-04
摘要： 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.
分类： 生物学 >> 植物学 >> 植物生物化学、植物生物物理学 提交时间： 2016-05-03
摘要： OsNRT1.1a is a low-affinity nitrate (NO3 ) transporter gene. In this study, another mRNA splicing product, OsNRT1.1b, putatively encoding a protein with six transmembrane domains, was identified based on the rice genomic database and bioinformatics analysis. OsNRT1.1a/OsNRT1.1b expression in Xenopus oocytes showed OsNRT1.1a-expressing oocytes accumulated 15N levels to about half as compared to OsNRT1.1b-expressing oocytes. The electrophysiological recording of OsNRT1.1b-expressing oocytes treated with 0.25mM NO3 confirmed 15N accumulation data. More functional assays were performed to examine the function of OsNRT1.1b in rice. The expression of both OsNRT1.1a and OsNRT1.1b was abundant in roots and downregulated by nitrogen (N) deficiency. The shoot biomass of transgenic rice plants with OsNRT1.1a or OsNRT1.1b overexpression increased under various N supplies under hydroponic conditions compared to wild-type (WT). The OsNRT1.1a overexpression lines showed increased plant N accumulation compared to the WT in 1.25mM NH4NO3 and 2.5mM NO3 – or NH4 þ treatments, but not in 0.125mMNH4NO3. However, OsNRT1.1b overexpression lines increased total N accumulation in all N treatments, including 0.125mM NH4NO3, suggesting that under low N condition, OsNRT1.1b would accumulate more N in plants and improve rice growth, but also that OsNRT1.1a had no such function in rice plants.(NO3) transporter gene. In this study, another mRNA splicing product, OsNRT1.1b, putatively encoding a protein with six transmembrane domains, was identified based on the rice genomic database and bioinformatics analysis. OsNRT1.1a/OsNRT1.1b expression in Xenopus oocytes showed OsNRT1.1a-expressing oocytes accumulated 15N levels to about half as compared to OsNRT1.1b-expressing oocytes. The electrophysiological recording of OsNRT1.1b-expressing oocytes treated with 0.25mM NO3 confirmed 15N accumulation data. More functional assays were performed to examine the function of OsNRT1.1b in rice. The expression of both OsNRT1.1a and OsNRT1.1b was abundant in roots and downregulated by nitrogen (N) deficiency. The shoot biomass of transgenic rice plants with OsNRT1.1a or OsNRT1.1b overexpression increased under various N supplies under hydroponic conditions compared to wild-type (WT). The OsNRT1.1a overexpression lines showed increased plant N accumulation compared to the WT in 1.25mM NH4NO3 and 2.5mM NO3 or NH4+ treatments, but not in 0.125mM NH4NO3. However, OsNRT1.1b overexpression lines increased total N accumulation in all N treatments, including 0.125mM NH4NO3, suggesting that under low N condition, OsNRT1.1b would accumulate more N in plants and improve rice growth, but also that OsNRT1.1a had no such function in rice plants.
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