The NO
3
-
transporter plays an important role in rice nitrogen acquisition and nitrogen-use efficiency. Our previous studies have shown that the high affinity systems for nitrate uptake in rice is mediated by a two-component NRT2/NAR2 transport system. In this study, transgenic plants were successful developed by overexpression of
OsNAR2.1
alone,
OsNRT2.3a
alone and co-overexpression of
OsNAR2.1
and
OsNRT2.3a
. Our field experiments indicated that transgenic lines expressing
p35S:OsNAR2.1
or
p35S:OsNAR2.1-p35S:OsNRT2.3a
constructs exhibited increased grain yields of approximately 14.1% and 24.6% compared with wild-type (cv. Wuyunjing 7, WT) plants, and the agricultural nitrogen use efficiency increased by 15.8% and 28.6%, respectively. Compared with WT, the
15
N influx in roots of
p35S:OsNAR2.1
and
p35S: OsNAR2.1-p35S:OsNRT2.3a
lines increased 18.9%‑27.8% in response to 0.2 mM, 2.5 mM
15
NO
3
–
, and 1.25 mM
15
NH
4
15
NO
3
, while there was no significant difference between
p35S:OsNAR2.1
and
p35S:OsNAR2.1-p35S:OsNRT2.3a
lines; only the
15
N distribution ratio of shoot to root for
p35S:OsNAR2.1-p35S:OsNRT2.3a
lines increased significantly. However, there were no significant differences in nitrogen use efficiency,
15
N influx in roots and the yield between the
p35S:NRT2.3a
transgenic lines and WT. This study indicated that co-overexpression of
OsNAR2.1
and
OsNRT2.3a
could increase rice yield and nitrogen use efficiency.