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Characterization and expression of Rubisco activase genes in Ipomoea batatas.

Jiang Y, Wang J, Tao X, Zhang Y - Mol. Biol. Rep. (2013)

Bottom Line: The results indicated that these two RCA isoforms may play different roles in regulating photosynthesis and they may be regulated by light, heat or both.In addition, there were interactions between Rubisco large subunit (RBCl) and short isoform RCA (RCAs) as well as RCAs and long isoform RCA (RCAl), but no interaction between RBCl and RCAl, implying they might form a sandwich-like structure (RBCl-RCAs-RCAl), at least in yeast cells.These results provided new information on the modulation of RCA genes in sweet potato, which could be useful in improving photosynthesis and plant growth in sweet potato.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Resource Biology and Eco-environment of Ministry of Education, Sichuan Key Laboratory of Molecular Biology and Biotechnology, College of Life Science, Sichuan University, Chengdu, 610064, People's Republic of China.

ABSTRACT
Two-dimensional electrophoresis, coupled with MALDI-TOF-MS, was used to identify differentially expressed proteins between young and mature leaves of sweet potato [Ipomoea batatas (L.) Lam]. The results showed that there were 25 differential proteins between young and mature leaves. The Rubisco activase (RCA) that catalyzes the activation of Rubisco in vivo and plays a crucial role in photosynthesis was among these 25 proteins. So far, little was known about the molecular biology of RCA in sweet potato. Here, this research reports the cloning and characterization of two genes encoding the short isoform and the long isoform of sweet potato RCAs. Analysis of DNA sequences of RCA suggested that the corresponding mRNAs were transcribed from two different genes. To study the roles of these two RCA isoforms in photosynthesis, we investigated the expression patterns of these RCA genes at the mRNA and protein levels every 2 h in a photoperiod and under different temperatures conditions. The results indicated that these two RCA isoforms may play different roles in regulating photosynthesis and they may be regulated by light, heat or both. In addition, there were interactions between Rubisco large subunit (RBCl) and short isoform RCA (RCAs) as well as RCAs and long isoform RCA (RCAl), but no interaction between RBCl and RCAl, implying they might form a sandwich-like structure (RBCl-RCAs-RCAl), at least in yeast cells. These results provided new information on the modulation of RCA genes in sweet potato, which could be useful in improving photosynthesis and plant growth in sweet potato.

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Structural features of the two Ib-RCA isoforms. a Nucleotide differences of the three Ib-RCAs homologous genes (named Ib-RCAs I, II, III) and the three Ib-RCAl homologous genes (named Ib-RCAl I, II, III). b Genomic structures of Ib-RCAs and Ib-RCAl genes. Exons (shown in gray boxes) and introns (shown in white lines) are drawn to scale
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Fig3: Structural features of the two Ib-RCA isoforms. a Nucleotide differences of the three Ib-RCAs homologous genes (named Ib-RCAs I, II, III) and the three Ib-RCAl homologous genes (named Ib-RCAl I, II, III). b Genomic structures of Ib-RCAs and Ib-RCAl genes. Exons (shown in gray boxes) and introns (shown in white lines) are drawn to scale

Mentions: In order to understand the relationship between the efficient photosynthesis and Rubisco activase in sweet potato, two isoforms of RCA genes were cloned and sequenced (Fig. 3). The sequencing results indicated that the two isoforms contained a 1,320 bp ORF (named Ib-RCAs) corresponding to a deduced protein of 440 amino acid residues and a 1,455 bp ORF (named Ib-RCAl) of a deduced protein of 485 amino acid residues, respectively. It was also found that, there were probably three highly homologous isoforms (homology >99 %) in both Ib-RCAs (named Ib-RCAs I, II, III) and Ib-RCAl (named Ib-RCAl I, II, III) (Fig. 3a). To ensure that these changes were not due to experimental errors, direct sequencing of PCR products of the full-length ORFs also confirmed the existence of three highly homologous isoforms with two peaks at the same base location (Fig. 3a). Although nucleotide sequences of three similar isoforms had different numbers of SNP, most of them occurred in the first or third base of codons, so amino acid sequences changed rarely (Supplementary Table S4).Fig. 3


Characterization and expression of Rubisco activase genes in Ipomoea batatas.

Jiang Y, Wang J, Tao X, Zhang Y - Mol. Biol. Rep. (2013)

Structural features of the two Ib-RCA isoforms. a Nucleotide differences of the three Ib-RCAs homologous genes (named Ib-RCAs I, II, III) and the three Ib-RCAl homologous genes (named Ib-RCAl I, II, III). b Genomic structures of Ib-RCAs and Ib-RCAl genes. Exons (shown in gray boxes) and introns (shown in white lines) are drawn to scale
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824211&req=5

Fig3: Structural features of the two Ib-RCA isoforms. a Nucleotide differences of the three Ib-RCAs homologous genes (named Ib-RCAs I, II, III) and the three Ib-RCAl homologous genes (named Ib-RCAl I, II, III). b Genomic structures of Ib-RCAs and Ib-RCAl genes. Exons (shown in gray boxes) and introns (shown in white lines) are drawn to scale
Mentions: In order to understand the relationship between the efficient photosynthesis and Rubisco activase in sweet potato, two isoforms of RCA genes were cloned and sequenced (Fig. 3). The sequencing results indicated that the two isoforms contained a 1,320 bp ORF (named Ib-RCAs) corresponding to a deduced protein of 440 amino acid residues and a 1,455 bp ORF (named Ib-RCAl) of a deduced protein of 485 amino acid residues, respectively. It was also found that, there were probably three highly homologous isoforms (homology >99 %) in both Ib-RCAs (named Ib-RCAs I, II, III) and Ib-RCAl (named Ib-RCAl I, II, III) (Fig. 3a). To ensure that these changes were not due to experimental errors, direct sequencing of PCR products of the full-length ORFs also confirmed the existence of three highly homologous isoforms with two peaks at the same base location (Fig. 3a). Although nucleotide sequences of three similar isoforms had different numbers of SNP, most of them occurred in the first or third base of codons, so amino acid sequences changed rarely (Supplementary Table S4).Fig. 3

Bottom Line: The results indicated that these two RCA isoforms may play different roles in regulating photosynthesis and they may be regulated by light, heat or both.In addition, there were interactions between Rubisco large subunit (RBCl) and short isoform RCA (RCAs) as well as RCAs and long isoform RCA (RCAl), but no interaction between RBCl and RCAl, implying they might form a sandwich-like structure (RBCl-RCAs-RCAl), at least in yeast cells.These results provided new information on the modulation of RCA genes in sweet potato, which could be useful in improving photosynthesis and plant growth in sweet potato.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Resource Biology and Eco-environment of Ministry of Education, Sichuan Key Laboratory of Molecular Biology and Biotechnology, College of Life Science, Sichuan University, Chengdu, 610064, People's Republic of China.

ABSTRACT
Two-dimensional electrophoresis, coupled with MALDI-TOF-MS, was used to identify differentially expressed proteins between young and mature leaves of sweet potato [Ipomoea batatas (L.) Lam]. The results showed that there were 25 differential proteins between young and mature leaves. The Rubisco activase (RCA) that catalyzes the activation of Rubisco in vivo and plays a crucial role in photosynthesis was among these 25 proteins. So far, little was known about the molecular biology of RCA in sweet potato. Here, this research reports the cloning and characterization of two genes encoding the short isoform and the long isoform of sweet potato RCAs. Analysis of DNA sequences of RCA suggested that the corresponding mRNAs were transcribed from two different genes. To study the roles of these two RCA isoforms in photosynthesis, we investigated the expression patterns of these RCA genes at the mRNA and protein levels every 2 h in a photoperiod and under different temperatures conditions. The results indicated that these two RCA isoforms may play different roles in regulating photosynthesis and they may be regulated by light, heat or both. In addition, there were interactions between Rubisco large subunit (RBCl) and short isoform RCA (RCAs) as well as RCAs and long isoform RCA (RCAl), but no interaction between RBCl and RCAl, implying they might form a sandwich-like structure (RBCl-RCAs-RCAl), at least in yeast cells. These results provided new information on the modulation of RCA genes in sweet potato, which could be useful in improving photosynthesis and plant growth in sweet potato.

Show MeSH
Related in: MedlinePlus