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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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Protein expression patterns in young leaves (a) and mature leaves (b) of sweet potato. Proteins were extracted from the young and mature leaves, separated by 2DE gels (24 cm IPG strip, pH3-10, 12.5 % SDS-PAGE), and stained by CBB-staining. The numbers indicate the differentially expressed proteins between young and mature leaves. The relative levels of protein expression were analyzed by imagemaster 5.0 software
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Fig1: Protein expression patterns in young leaves (a) and mature leaves (b) of sweet potato. Proteins were extracted from the young and mature leaves, separated by 2DE gels (24 cm IPG strip, pH3-10, 12.5 % SDS-PAGE), and stained by CBB-staining. The numbers indicate the differentially expressed proteins between young and mature leaves. The relative levels of protein expression were analyzed by imagemaster 5.0 software

Mentions: Proteomic characterization of sweet potato leaves was achieved by 2-DE analysis of proteins extracted from leaves at young and mature stages. Representative gels were shown in Fig. 1. There was 806 and 753 protein spots detected in young and mature leaves, respectively. Relative intensity analysis of protein spots by Imagemaster 5.0 software displayed that 565 protein points were matched and the related coefficient was 0.798 (data not shown). To evaluate the differentially expressed proteins, we performed statistical analysis with these data using the student T test and found 25 differentially expressed proteins with good reproducibility (the differences were more than 2.5-fold; p < 0.05). Seventeen out of 25 proteins were up-regulated, and the other 8 were down-regulated (Fig. 1). All 25 proteins were excised from gels, digested and subjected to MALDI-TOF–TOF/MS analysis. Since there was no available genome sequence of sweet potato, identification of sweet potato spots by MALDI-TOF–TOF-MS experiments proved challenging. Therefore, homology searches were carried out in NCBI GreenPlants database and the transcriptomic database of sweet potato [31]. The functions of 20 proteins were known, but the other five proteins could not be identified. Among the 20 known proteins, 13 were up-regulated and 7 were down-regulated (Table 1). Based on their functions, these 20 proteins were classified into six categories: photosynthesis, energy metabolism, transcription regulation, stress-responsive, skeleton protein, and transport protein (Supplementary Table S3).Fig. 1


Characterization and expression of Rubisco activase genes in Ipomoea batatas.

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

Protein expression patterns in young leaves (a) and mature leaves (b) of sweet potato. Proteins were extracted from the young and mature leaves, separated by 2DE gels (24 cm IPG strip, pH3-10, 12.5 % SDS-PAGE), and stained by CBB-staining. The numbers indicate the differentially expressed proteins between young and mature leaves. The relative levels of protein expression were analyzed by imagemaster 5.0 software
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig1: Protein expression patterns in young leaves (a) and mature leaves (b) of sweet potato. Proteins were extracted from the young and mature leaves, separated by 2DE gels (24 cm IPG strip, pH3-10, 12.5 % SDS-PAGE), and stained by CBB-staining. The numbers indicate the differentially expressed proteins between young and mature leaves. The relative levels of protein expression were analyzed by imagemaster 5.0 software
Mentions: Proteomic characterization of sweet potato leaves was achieved by 2-DE analysis of proteins extracted from leaves at young and mature stages. Representative gels were shown in Fig. 1. There was 806 and 753 protein spots detected in young and mature leaves, respectively. Relative intensity analysis of protein spots by Imagemaster 5.0 software displayed that 565 protein points were matched and the related coefficient was 0.798 (data not shown). To evaluate the differentially expressed proteins, we performed statistical analysis with these data using the student T test and found 25 differentially expressed proteins with good reproducibility (the differences were more than 2.5-fold; p < 0.05). Seventeen out of 25 proteins were up-regulated, and the other 8 were down-regulated (Fig. 1). All 25 proteins were excised from gels, digested and subjected to MALDI-TOF–TOF/MS analysis. Since there was no available genome sequence of sweet potato, identification of sweet potato spots by MALDI-TOF–TOF-MS experiments proved challenging. Therefore, homology searches were carried out in NCBI GreenPlants database and the transcriptomic database of sweet potato [31]. The functions of 20 proteins were known, but the other five proteins could not be identified. Among the 20 known proteins, 13 were up-regulated and 7 were down-regulated (Table 1). Based on their functions, these 20 proteins were classified into six categories: photosynthesis, energy metabolism, transcription regulation, stress-responsive, skeleton protein, and transport protein (Supplementary Table S3).Fig. 1

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