a1_Two full-length cDNAs (SGrca1 and SGrca2) encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase (RCA) were cloned from a heterophyllous aquatic plant, Sagittaria graminea, using Rapid-Amplification of cDNA Ends (RACE). SGrca1 contains a 1,320 bp open reading frame encoding a protein of 440 amino acids, and SGrca2 is exactly identical to SGrca1 except for 330 bp missing in the middle of SGrca1. Sequence analysis of cDNA and genomic DNA indicated both two cDNAs were generated from a common gene via alternative splicing. The deduced amino acid sequence encoded by SGrca1 showed 75-82% identity with other RCAs from higher plants and showed high homology in three highly conserved motifs associated with ATP-binding sites. RT-PCR analysis suggested both SGrca1 and SGrca2 were expressed in green tissues. During a 14 h light/10 h dark photoperiod, both aerial and submerged leaves exhibited the similar expression pattern of SGrca1 and SGrca2 with SGrca1 as the dominant form, but the accumulation of both SGrca1 and SGrca2 mRNA was significantly inhibited in the submerged leaves., a2_Western blot analysis showed that both SGrca1 and SGrca2 had their translation products, the 43 kDa form and the 31 kDa form expressing in leaves. Interestingly, the aerial leaves expressed higher amount of the 43 kDa form compared with the 31 kDa form, while it was reversed in the submerged leaves. The results demonstrated that both environments regulated the RCA gene expression at both transcriptional and posttranscriptional level. In addition, co-immunoprecipitation assay revealed that the isolated Rubisco-RCA complex contained both the 43 and 31 kDa forms, and the proportion of the 31 kDa form was obviously enhanced in the submerged leaves. The results indicated that both the 43 kDa and 31 kDa forms were involved in Rubisco and RCA interaction and the increased incorporation of the 31 kDa form was associated with submerged photosynthetic environment., D. Wang, S. Z. Xie, J. Yang, Q. F. Wang., and Obsahuje bibliografii
To investigate into the relationship between two Rubisco activase (RCA) isoforms and photosynthetic rate, a set of enzyme-linked immunosorbent assay (ELISA) were developed for accurate quantification of two RCA polypeptides based on two specific monoclonal antibodies against different RCA isoforms. The results showed that content of RCA small isoform (RCAS) was 5-fold more than that of RCA large isoform (RCAL) content in all leaves and the RCAL/RCAS ratio reached maximum in the leaf with the highest photosynthetic rate. Although the difference in two RCA polypeptides accumulation in leaves was caused by different transcript level of two isoforms, the decrease of RCAL/RCAS ratio during leaf aging was not attributed to transcriptional regulation. The leaves with higher photosynthetic capacity exhibited higher RCAL/RCAS ratio and the decrease in photosynthetic rate and Rubisco activation state highly correlated with the decline of RCAL/RCAS ratio during leaf aging. Our results suggest that there is a posttranscriptional mechanism regulating the RCAL/RCAS ratio, which may play as a regulator modulating photosynthetic capacity during leaf aging in rice plant. and D. Wang ... [et al.].
To investigate how excess excitation energy is dissipated in a ribulose-1,5-bisphospate carboxylase/oxygenase activase antisense transgenic rice with net photosynthetic rate (PN) half of that of wild type parent, we measured the response curve of PN to intercellular CO2 concentration (Ci), electron transport rate (ETR), quantum yield of open photosystem 2 (PS2) reaction centres under irradiation (Fv'/Fm'), efficiency of total PS2 centres (ΦPS2), photochemical (qP) and non-photochemical quenching (NPQ), post-irradiation transient increase in chlorophyll (Chl) fluorescence (PITICF), and P700+ re-reduction. Carboxylation efficiency dependence on Ci, ETR at saturation irradiance, and Fv'/Fm', ΦPS2, and qP under the irradiation were significantly lower in the mutant. However, NPQ, energy-dependent quenching (qE), PITICF, and P700+ re-reduction were significantly higher in the mutant. Hence the mutant down-regulates linear ETR and stimulates cyclic electron flow around PS1, which may generate the ΔpH to support NPQ and qE for dissipation of excess excitation energy. and S.-H. Jin ... [et al.].