Changes in nitrogen assimilation, metabolism, and growth in transgenic rice plants expressing a fungal NADP(H)-dependent glutamate dehydrogenase (gdhA).
In plants, glutamine synthetase (GS) is the enzyme that is mainly responsible for the assimilation of ammonium. Conversely, in microorganisms such as bacteria and Ascomycota, NADP(H)-dependent glutamate dehydrogenase (GDH) and GS both have important roles in ammonium assimilation. Here, we report the changes in nitrogen assimilation, metabolism, growth, and grain yield of rice plants caused by an ectopic expression of NADP(H)-GDH (gdhA) from the fungus Aspergillus niger in the cytoplasm. An investigation of the kinetic properties of purified recombinant protein showed that the fungal gdhA had 5.4-10.2 times higher V(max) value and 15.9-43.1 times higher K(m) value for NH(4)(+), compared with corresponding values for rice cytosolic GS as reported in the literature. These results suggested that the introduction of fungal GDH into rice could modify its ammonium assimilation pathway. We therefore expressed gdhA in the cytoplasm of rice plants. NADP(H)-GDH activities in the gdhA-transgenic lines were markedly higher than those in a control line. Tracer experiments by feeding with (15)NH(4)(+) showed that the introduced gdhA, together with the endogenous GS, directly assimilated NH(4)(+) absorbed from the roots. Furthermore, in comparison with the control line, the transgenic lines showed an increase in dry weight and nitrogen content when sufficient nitrogen was present, but did not do so under low-nitrogen conditions. Under field condition, the transgenic line examined showed a significant increase in grain yield in comparison with the control line. These results suggest that the introduction of fungal gdhA into rice plants could lead to better growth and higher grain yield by enhancing the assimilation of ammonium.
Abiko
Tomomi
T
Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
Wakayama
Masataka
M
Kawakami
Akira
A
Obara
Mitsuhiro
M
Kisaka
Hiroaki
H
Miwa
Tetsuya
T
Aoki
Naohiro
N
Ohsugi
Ryu
R
D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D018086:Glutamate Dehydrogenase (NADP+) / Q000235:genetics / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism*; D012275:Oryza / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*
D005656:Fungal Proteins; D018086:Glutamate Dehydrogenase (NADP+); D009584:Nitrogen