title: Small subunit of a cold-resistant plant, Timothy, does not significantly alter the catalytic properties of Rubisco in transgenic rice.
abstract: Effects of overexpression of high activity-type Rubisco small subunit (RbcS) from a cold-resistant plant, timothy (Phleum pratense), on kinetic properties of Rubisco were studied in rice (Oryza sativa). The full-length mRNA sequence of timothy RbcS (PpRbcS1) was determined by 5'RACE and 3'RACE. The coding sequence of PpRbcS1 was fused to the chlorophyll a/b-binding protein promoter and introduced into rice. PpRbcS was highly expressed in leaf blade and accounted for approximately 30 % of total RbcS in homozygous transgenic lines. However, the catalytic turnover rate and K m for CO2 of Rubisco did not significantly change in these transgenic lines compared to non-transgenic rice, suggesting that PpRbcS1 is not effective for improvement of catalytic efficiency of rice Rubisco. The photosynthetic rate and growth were essentially unchanged, whereas the photosynthetic rate at low CO2 condition was marginally increased in transgenic lines. Rubisco content was significantly increased, whereas soluble protein, nitrogen, and chlorophyll contents were unchanged in transgenic lines compared to non-transgenic rice. Because the kinetic properties were similar, observed slight increase in photosynthetic rate at low CO2 is considered to be large due to increase in Rubisco content in transgenic lines. Introduction of foreign RbcS is an effective approach for the improvement of Rubisco kinetics and photosynthesis. However, in this study, it was suggested that RbcS of high activity-type Rubisco, even showing higher amino acid identity with rice RbcS, did not always enhance the catalytic turnover rate of Rubisco in rice. Thus, we should carefully select RbcS to be overexpressed before introduction.
authors.affiliation: Graduate School of Agricultural Science, Laboratory of Crop Science, Kobe University, Nada-ku, Kobe, 657-8501, Japan, fukayama@people.kobe-u.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Fukayama
authors.forename: Atsushi
authors.initials: A
authors.lastname: Koga
authors.forename: Tomoko
authors.initials: T
authors.lastname: Hatanaka
authors.forename: Shuji
authors.initials: S
authors.lastname: Misoo
mesh_terms: D000595:Amino Acid Sequence; D055162:Biocatalysis; D002734:Chlorophyll / Q000378:metabolism; D003080:Cold Temperature; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism; D012275:Oryza / Q000235:genetics*; D031784:Phleum / Q000201:enzymology*; D010788:Photosynthesis; D030821:Plants, Genetically Modified; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism*; D012273:Ribulose-Bisphosphate Carboxylase / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment
chemical_list: D021122:Protein Subunits; D002734:Chlorophyll; D012273:Ribulose-Bisphosphate Carboxylase; D009584:Nitrogen