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Table 1.

The characteristics of primers used for quantitative real-time PCR in C. sinensis.

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Fig 1.

Confirmation of primer specificity and amplicon size.

(a) Melting curve analysis of 12 candidate reference genes. (b) Amplification results for 12 candidate genes using a C. sinensis cDNA template. M: DL2000 DNA Marker.

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Fig 2.

Quantification cycle (Cq) values of the 12 candidate reference genes in C. sinensis leaves under metal stresses.

The lines across boxes represent the mean Cq values. The boxes indicate the 25th and 75th percentiles, while the whiskers correspond to the maximum and minimum values.

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Fig 3.

Determination of the optimal number of reference genes required for effective data normalization.

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Table 2.

Gene expression stability ranked by geNorm, NormFinder, and BestKeeper software programs.

SD: standard deviation; CV: coefficient of variation.

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Fig 4.

Relative quantification of CsPCS1 gene expression using candidate reference genes in Al-stressed C. sinensis leaves.

Data are presented as the means ± standard deviation of four replicates. Significant differences were determined by Duncan’s multiple range test (* P < 0.05, ** P < 0.01, and ** P < 0.001).

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