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Identification of Rubisco rbcL and rbcS in Camellia oleifera and their potential as molecular markers for selection of high tea oil cultivars.

Chen Y, Wang B, Chen J, Wang X, Wang R, Peng S, Chen L, Ma L, Luo J - Front Plant Sci (2015)

Bottom Line: The Co-rbcL expression in 'Xianglin 14' was significantly higher than 'Xianglin 1', and 'Xianglin 1' was greater than 'Hengchong 89'.The expression levels of Co-rbcS in 'Xianglin 1' and 'Xianglin 14' were similar but were significantly greater than in 'Hengchong 89'.In combination with the measurement of net photosynthetic rates, the early identification of potential high oil production cultivars would significantly shorten plant breeding time and increase breeding efficiency.

View Article: PubMed Central - PubMed

Affiliation: National Engineering Technology Research Center of Oil-tea Camellia, Hunan Academy of Forestry Changsha, China.

ABSTRACT
Tea oil derived from seeds of Camellia oleifera Abel. is high-quality edible oil in China. This study isolated full-length cDNAs of Rubisco subunits rbcL and rbcS from C. oleifera. The rbcL has 1,522 bp with a 1,425 bp coding region, encoding 475 amino acids; and the rbcS has 615 bp containing a 528 bp coding region, encoding 176 amino acids. The expression level of the two genes, designated as Co-rbcL and Co-rbcS, was determined in three C. oleifera cultivars: Hengchong 89, Xianglin 1, and Xianglin 14 whose annual oil yields were 546.9, 591.4, and 657.7 kg ha(-1), respectively. The Co-rbcL expression in 'Xianglin 14' was significantly higher than 'Xianglin 1', and 'Xianglin 1' was greater than 'Hengchong 89'. The expression levels of Co-rbcS in 'Xianglin 1' and 'Xianglin 14' were similar but were significantly greater than in 'Hengchong 89'. The net photosynthetic rate of 'Xianglin 14' was significantly higher than 'Xianglin 1', and 'Xianglin 1' was higher than 'Hengchong 89'. Pearson's correlation analysis showed that seed yields and oil yields were highly correlated with the expression level of Co-rbcL at P < 0.001 level; and the expression of Co-rbcS was correlated with oil yield at P < 0.01 level. Net photosynthetic rate was also correlated with oil yields and seed yields at P < 0.001 and P < 0.01 levels, respectively. Our results suggest that Co-rbcS and Co-rbcL in particular could potentially be molecular markers for early selection of high oil yield cultivars. In combination with the measurement of net photosynthetic rates, the early identification of potential high oil production cultivars would significantly shorten plant breeding time and increase breeding efficiency.

No MeSH data available.


Related in: MedlinePlus

Polymerase chain reaction cloning of rbcS from leaves of C. oleifera and its recombinant strains. (A) The degenerate PCR amplification fragment of rbcS in lane 1. (B) Lanes 1, 2, and 3 are positive recombinants of rbcS. (C) Lane 1 indicates the 5′ RACER result of Co-rbcS. (D) Lanes 1–6 were positive recombinants of the 5′ RACER of Co-rbcS. (E) Lane 1 is the 3′ RACER result of Co-rbcS. (F) Lanes 1–10 are positive clones of Co-rbcS. M represents 100 bp plus DNA ladder.
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Figure 4: Polymerase chain reaction cloning of rbcS from leaves of C. oleifera and its recombinant strains. (A) The degenerate PCR amplification fragment of rbcS in lane 1. (B) Lanes 1, 2, and 3 are positive recombinants of rbcS. (C) Lane 1 indicates the 5′ RACER result of Co-rbcS. (D) Lanes 1–6 were positive recombinants of the 5′ RACER of Co-rbcS. (E) Lane 1 is the 3′ RACER result of Co-rbcS. (F) Lanes 1–10 are positive clones of Co-rbcS. M represents 100 bp plus DNA ladder.

Mentions: A partial sequence of rbcS gene was first obtained from degenerate RT-PCR reactions. Sequence analysis showed that this fragment had 324 bp (Figures 4A,B) encoding a deduced peptide sequence of 122 amino acids. This sequence shared 94% homology with rbcS of C. sinensis (GenBank accession no. EF011075), suggesting that it was a fragment of rbcS gene. Based on this fragment, a set of gene specific primers were designed in order to obtain the 5′ and 3′ ends (Figures 4C–F). The resultant full-length rbcS gene was found to be 615 bp. This sequence had an open reading frame of 528 bp starting with an initiation codon ATG at position of the 2nd and 4th nucleotides and ending with a termination codon TAA at position of the 530th and 532nd nucleotides, encoding 176 amino acid residues with a molecular mass of 19.69 kDa. The GC content of this sequence was about 49.15%. The predicted nucleotide sequence of the coding region shared more than 95% homology with that of rbcS mRNA from C. sinensis (GenBank accession no. EF011075). This sequence was designated as Co-rbcS, and has been deposited in GenBank with an accession no. KJ721196.


Identification of Rubisco rbcL and rbcS in Camellia oleifera and their potential as molecular markers for selection of high tea oil cultivars.

Chen Y, Wang B, Chen J, Wang X, Wang R, Peng S, Chen L, Ma L, Luo J - Front Plant Sci (2015)

Polymerase chain reaction cloning of rbcS from leaves of C. oleifera and its recombinant strains. (A) The degenerate PCR amplification fragment of rbcS in lane 1. (B) Lanes 1, 2, and 3 are positive recombinants of rbcS. (C) Lane 1 indicates the 5′ RACER result of Co-rbcS. (D) Lanes 1–6 were positive recombinants of the 5′ RACER of Co-rbcS. (E) Lane 1 is the 3′ RACER result of Co-rbcS. (F) Lanes 1–10 are positive clones of Co-rbcS. M represents 100 bp plus DNA ladder.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4379756&req=5

Figure 4: Polymerase chain reaction cloning of rbcS from leaves of C. oleifera and its recombinant strains. (A) The degenerate PCR amplification fragment of rbcS in lane 1. (B) Lanes 1, 2, and 3 are positive recombinants of rbcS. (C) Lane 1 indicates the 5′ RACER result of Co-rbcS. (D) Lanes 1–6 were positive recombinants of the 5′ RACER of Co-rbcS. (E) Lane 1 is the 3′ RACER result of Co-rbcS. (F) Lanes 1–10 are positive clones of Co-rbcS. M represents 100 bp plus DNA ladder.
Mentions: A partial sequence of rbcS gene was first obtained from degenerate RT-PCR reactions. Sequence analysis showed that this fragment had 324 bp (Figures 4A,B) encoding a deduced peptide sequence of 122 amino acids. This sequence shared 94% homology with rbcS of C. sinensis (GenBank accession no. EF011075), suggesting that it was a fragment of rbcS gene. Based on this fragment, a set of gene specific primers were designed in order to obtain the 5′ and 3′ ends (Figures 4C–F). The resultant full-length rbcS gene was found to be 615 bp. This sequence had an open reading frame of 528 bp starting with an initiation codon ATG at position of the 2nd and 4th nucleotides and ending with a termination codon TAA at position of the 530th and 532nd nucleotides, encoding 176 amino acid residues with a molecular mass of 19.69 kDa. The GC content of this sequence was about 49.15%. The predicted nucleotide sequence of the coding region shared more than 95% homology with that of rbcS mRNA from C. sinensis (GenBank accession no. EF011075). This sequence was designated as Co-rbcS, and has been deposited in GenBank with an accession no. KJ721196.

Bottom Line: The Co-rbcL expression in 'Xianglin 14' was significantly higher than 'Xianglin 1', and 'Xianglin 1' was greater than 'Hengchong 89'.The expression levels of Co-rbcS in 'Xianglin 1' and 'Xianglin 14' were similar but were significantly greater than in 'Hengchong 89'.In combination with the measurement of net photosynthetic rates, the early identification of potential high oil production cultivars would significantly shorten plant breeding time and increase breeding efficiency.

View Article: PubMed Central - PubMed

Affiliation: National Engineering Technology Research Center of Oil-tea Camellia, Hunan Academy of Forestry Changsha, China.

ABSTRACT
Tea oil derived from seeds of Camellia oleifera Abel. is high-quality edible oil in China. This study isolated full-length cDNAs of Rubisco subunits rbcL and rbcS from C. oleifera. The rbcL has 1,522 bp with a 1,425 bp coding region, encoding 475 amino acids; and the rbcS has 615 bp containing a 528 bp coding region, encoding 176 amino acids. The expression level of the two genes, designated as Co-rbcL and Co-rbcS, was determined in three C. oleifera cultivars: Hengchong 89, Xianglin 1, and Xianglin 14 whose annual oil yields were 546.9, 591.4, and 657.7 kg ha(-1), respectively. The Co-rbcL expression in 'Xianglin 14' was significantly higher than 'Xianglin 1', and 'Xianglin 1' was greater than 'Hengchong 89'. The expression levels of Co-rbcS in 'Xianglin 1' and 'Xianglin 14' were similar but were significantly greater than in 'Hengchong 89'. The net photosynthetic rate of 'Xianglin 14' was significantly higher than 'Xianglin 1', and 'Xianglin 1' was higher than 'Hengchong 89'. Pearson's correlation analysis showed that seed yields and oil yields were highly correlated with the expression level of Co-rbcL at P < 0.001 level; and the expression of Co-rbcS was correlated with oil yield at P < 0.01 level. Net photosynthetic rate was also correlated with oil yields and seed yields at P < 0.001 and P < 0.01 levels, respectively. Our results suggest that Co-rbcS and Co-rbcL in particular could potentially be molecular markers for early selection of high oil yield cultivars. In combination with the measurement of net photosynthetic rates, the early identification of potential high oil production cultivars would significantly shorten plant breeding time and increase breeding efficiency.

No MeSH data available.


Related in: MedlinePlus