Cloning and sequencing of a full-length cDNA encoding the rubpcase small subunit (RbcS) in tea (Camellia sinensis).

This study was aimed to isolate ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) from tea plant [Camellia sinensis (L.) O. Kuntze]. In the study of transcriptional profiling of gene expression from tea flower bud development stage by cDNA-AFLP (cDNA amplified fragment length polymorphism), we have isolated some transcript-derived fragments (TDFs) occurring in both the young and mature flower bud. One of them showed a high degree of similarity to RbcS. Based on the fragment, the full length of RbcS with 769-bp (EF011075) cDNA was obtained via rapid amplification of cDNA ends (RACE). It contained an open reading frame of 176 amino acids consisting of a chloroplast transit peptide with 52 amino acids and a mature protein of 124 amino acids. The amino acids sequence presented a high identity to those of other plant RbcS genes. It also contains three conserved domains and a protein kinase C phosphorylation site, one tyrosine kinase phosphorylation site and two N-myristoylation sites. Analysis by RT-PCR showed that the expression of RbcS in tea from high to low was leaf, young stem, young flower bud and mature flower bud, respectively. The isolation of the tea Rubisco small subunit gene establishes a good foundation for further study on the photosynthesis of tea plant.

[1]  W. Gruissem,et al.  Organ-Specific Differential Regulation of a Promoter Subfamily for the Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Small Subunit Genes in Tomato , 1995, Plant physiology.

[2]  G. Coruzzi,et al.  Tissue‐specific and light‐regulated expression of a pea nuclear gene encoding the small subunit of ribulose‐1,5‐bisphosphate carboxylase. , 1984, The EMBO journal.

[3]  J. Langdale,et al.  Transcripts of maize RbcS genes accumulate differentially in C3 and C4 tissues , 1998, Plant Molecular Biology.

[4]  M. Kozak.  Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes , 1986, Cell.

[5]  W. Gruissem,et al.  Expression dynamics of the tomato rbcS gene family during development. , 1991, The Plant cell.

[6]  M. Kozak,et al.  Recognition of AUG and alternative initiator codons is augmented by G in position +4 but is not generally affected by the nucleotides in positions +5 and +6 , 1997, The EMBO journal.

[7]  Hardy Pfanz,et al.  Non-foliar photosynthesis – a strategy of additional carbon acquisition , 2003 .

[8]  Wang Xiaochun.  Analysis of Differential Gene Expression of Flower bud of Tea Plant (Camellia sinensis) by cDNA-AFLP , 2007 .

[9]  P. Maliga,et al.  Plastome engineering of ribulose-1,5-bisphosphate carboxylase/oxygenase in tobacco to form a sunflower large subunit and tobacco small subunit hybrid. , 1999, Plant physiology.

[10]  R. J. Spreitzer.  Questions about the complexity of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase , 1999, Photosynthesis Research.

[11]  A. Thomson,et al.  Identification of a Novel AU-Rich Element in the 3′ Untranslated Region of Epidermal Growth Factor Receptor mRNA That Is the Target for Regulated RNA-Binding Proteins , 2001, Molecular and Cellular Biology.

[12]  C. Bachem,et al.  Transcript Imaging with cDNA-AFLP: A Step-by-Step Protocol , 1998, Plant Molecular Biology Reporter.

[13]  T. Sasanuma.  Characterization of the rbcS multigene family in wheat: subfamily classification, determination of chromosomal location and evolutionary analysis , 2001, Molecular Genetics and Genomics.

[14]  R. J. Spreitzer,et al.  Role of the small subunit in ribulose-1,5-bisphosphate carboxylase/oxygenase. , 2003, Archives of biochemistry and biophysics.

[15]  A. Dhingra,et al.  Enhanced translation of a chloroplast-expressed RbcS gene restores small subunit levels and photosynthesis in nuclear RbcS antisense plants. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[16]  S. Rodermel.  Subunit control of Rubisco biosynthesis – a relic of an endosymbiotic past? , 1999, Photosynthesis Research.

[17]  B. Read,et al.  A hybrid ribulosebisphosphate carboxylase/oxygenase enzyme exhibiting a substantial increase in substrate specificity factor. , 1992, Biochemistry.

[18]  R. Van der Hoeven,et al.  Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: analysis of gene expression during potato tuber development. , 1996, The Plant journal : for cell and molecular biology.

[19]  A. Makino,et al.  Photosynthesis and Ribulose 1,5-Bisphosphate Carboxylase in Rice Leaves: Changes in Photosynthesis and Enzymes Involved in Carbon Assimilation from Leaf Development through Senescence. , 1983, Plant physiology.

[20]  H. Bohnert,et al.  Identification of an assembly domain in the small subunit of ribulose-1,5-bisphosphate carboxylase. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[21]  I. Andersson,et al.  Reexamination of the Three-Dimensional Structure of the Small Subunit of RuBisCo from Higher Plants , 1989, Science.

[22]  C. Dean,et al.  Structure, evolution and regulation of RbcS genes in higher plants , 1989 .