title: Asparagine synthetase1, but not asparagine synthetase2, is responsible for the biosynthesis of asparagine following the supply of ammonium to rice roots.
abstract: Asparagine is synthesized from glutamine by the reaction of asparagine synthetase (AS) and is the major nitrogen form in both xylem and phloem sap in rice (Oryza sativa L.). There are two genes encoding AS, OsAS1 and OsAS2, in rice, but the functions of individual AS isoenzymes are largely unknown. Cell type- and NH4(+)-inducible expression of OsAS1 as well as analyses of knockout mutants were carried out in this study to characterize AS1. OsAS1 was mainly expressed in the roots, with in situ hybridization showing that the corresponding mRNA was specifically accumulated in the three cell layers of the root surface (epidermis, exodermis and sclerenchyma) in an NH4(+)-dependent manner. Conversely, OsAS2 mRNA was abundant in leaf blades and sheathes of rice. Although OsAS2 mRNA was detectable in the roots, its content decreased when NH4(+) was supplied. Retrotransposon-mediated knockout mutants lacking AS1 showed slight stimulation of shoot length and slight reduction in root length at the seedling stage. On the other hand, the mutation caused an approximately 80-90% reduction in free asparagine content in both roots and xylem sap. These results suggest that AS1 is responsible for the synthesis of asparagine in rice roots following the supply of NH4(+). Characteristics of the NH4(+)-dependent increase and the root surface cell-specific expression of OsAS1 gene are very similar to our previous results on cytosolic glutamine synthetase1;2 and NADH-glutamate synthase1 in rice roots. Thus, AS1 is apparently coupled with the primary assimilation of NH4(+) in rice roots.
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan.
authors: Miwa
authors: M
authors: Ohashi
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan.
authors: Keiki
authors: K
authors: Ishiyama
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan.
authors: Soichi
authors: S
authors: Kojima
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan.
authors: Noriyuki
authors: N
authors: Konishi
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan Present address: Cambridge Technology Partners Co. Ltd., 1-1-1 Toyosu, Koto-ku, Tokyo 135-8560 Japan.
authors: Kentaro
authors: K
authors: Nakano
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan.
authors: Keiichi
authors: K
authors: Kanno
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan.
authors: Toshihiko
authors: T
authors: Hayakawa
authors: Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Aoba-ku, Sendai, 981-8555 Japan tyamaya@biochem.tohoku.ac.jp.
authors: Tomoyuki
authors: T
authors: Yamaya
mesh_terms: D000595:Amino Acid Sequence; D064751:Ammonium Compounds / Q000494:pharmacology*; D001216:Asparagine / Q000096:biosynthesis*; D001217:Aspartate-Ammonia Ligase / Q000737:chemistry / Q000378:metabolism*; D002244:Carbon / Q000378:metabolism; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D055786:Gene Knockout Techniques; D017343:Genes, Plant; D018519:Meristem / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009584:Nitrogen / Q000378:metabolism; D012275:Oryza / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D010641:Phenotype; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism*; D018517:Plant Roots / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D018520:Plant Shoots / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D036226:Seedlings / Q000187:drug effects / Q000235:genetics
chemical_list: D064751:Ammonium Compounds; D010940:Plant Proteins; D012333:RNA, Messenger; D001216:Asparagine; D002244:Carbon; D001217:Aspartate-Ammonia Ligase; D009584:Nitrogen
keywords: Ammonium assimilation; Asparagine synthetase; Cytosolic glutamine synthetase; Knockout mutant; NADH-glutamate synthase; Rice root