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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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Related in: MedlinePlus

Semi-quantitative RT-PCR analysis of genes differentially expressed in young and mature leaves. The products of RT-PCR were separated on 1.2 % agarose gel. Spot 5, oxygen evolving enhancer protein 2; Spot 7, fructose-bisphosphate aldolase; Spot 11/12, Rubisco activase; Spot 13, nucleoside diphosphate kinase; Spot 17, elongation factor-1 α subunit; Spot 19, granule bound starch synthase; Spot 20, cyclophilin; Spot 21, carbonic anhydrase; Spot 25, phosphoribulokinase. The β-actin gene was used as an internal control
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Fig2: Semi-quantitative RT-PCR analysis of genes differentially expressed in young and mature leaves. The products of RT-PCR were separated on 1.2 % agarose gel. Spot 5, oxygen evolving enhancer protein 2; Spot 7, fructose-bisphosphate aldolase; Spot 11/12, Rubisco activase; Spot 13, nucleoside diphosphate kinase; Spot 17, elongation factor-1 α subunit; Spot 19, granule bound starch synthase; Spot 20, cyclophilin; Spot 21, carbonic anhydrase; Spot 25, phosphoribulokinase. The β-actin gene was used as an internal control

Mentions: To investigate whether the proteins were correlated with their transcript levels, nine proteins with significant differences were chosen for further study by semi-quantitative RT-PCR. As shown in Fig. 2, the proteins and mRNAs appear to be generally correlated. However, there also were some variations at the protein and mRNA levels. For example, the expression of spot 13 (nucleoside diphosphate kinase), spot 20 (cyclophilin), spot 25 (phosphoribulokinase) in young leaves were about −9.5, −6.4, 2.6 times higher than those of the mature leaves at protein level, while at the mRNA level, the differences between the young and mature leaves were not striking (Figs. 1, 2).Fig. 2


Characterization and expression of Rubisco activase genes in Ipomoea batatas.

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

Semi-quantitative RT-PCR analysis of genes differentially expressed in young and mature leaves. The products of RT-PCR were separated on 1.2 % agarose gel. Spot 5, oxygen evolving enhancer protein 2; Spot 7, fructose-bisphosphate aldolase; Spot 11/12, Rubisco activase; Spot 13, nucleoside diphosphate kinase; Spot 17, elongation factor-1 α subunit; Spot 19, granule bound starch synthase; Spot 20, cyclophilin; Spot 21, carbonic anhydrase; Spot 25, phosphoribulokinase. The β-actin gene was used as an internal control
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig2: Semi-quantitative RT-PCR analysis of genes differentially expressed in young and mature leaves. The products of RT-PCR were separated on 1.2 % agarose gel. Spot 5, oxygen evolving enhancer protein 2; Spot 7, fructose-bisphosphate aldolase; Spot 11/12, Rubisco activase; Spot 13, nucleoside diphosphate kinase; Spot 17, elongation factor-1 α subunit; Spot 19, granule bound starch synthase; Spot 20, cyclophilin; Spot 21, carbonic anhydrase; Spot 25, phosphoribulokinase. The β-actin gene was used as an internal control
Mentions: To investigate whether the proteins were correlated with their transcript levels, nine proteins with significant differences were chosen for further study by semi-quantitative RT-PCR. As shown in Fig. 2, the proteins and mRNAs appear to be generally correlated. However, there also were some variations at the protein and mRNA levels. For example, the expression of spot 13 (nucleoside diphosphate kinase), spot 20 (cyclophilin), spot 25 (phosphoribulokinase) in young leaves were about −9.5, −6.4, 2.6 times higher than those of the mature leaves at protein level, while at the mRNA level, the differences between the young and mature leaves were not striking (Figs. 1, 2).Fig. 2

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