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Table 1.

Primers used to amplify partial sequences of 7 reference genes and miR159 from cucumber samples.

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Fig 1.

RT-qPCR cycle threshold values of seven reference genes in cucumber leaf, stem and root samples at different time points post-inoculation with CGMMV.

Error bars represent the mean of three technical replicates ± SD.

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Table 2.

Expression stability of seven reference genes in cucumber analyzed using the delta-Ct algorithm.

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Fig 2.

GeNorm data for 5 cucumber reference genes in cucumber leaf, stem and root samples at different time points post-inoculation with CGMMV.

(A) to (D) Average expression stability values (M) of cucumber reference genes during stepwise exclusion of the least stable reference gene in different tissue samples. (A) leaf, (B) stem, (C) root, and (D) overall. (E) to (H) Optimal number of reference genes required for accurate normalization of expression based on pairwise variation (Vn/n+1) analysis of the normalization factors of the reference genes in different tissue samples. (E) leaf, (F) stem, (G) root, and (H) overall.

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Table 3.

Expression stability of seven reference genes in cucumber analyzed using the NormFinder algorithm.

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Table 4.

Most suitable combination of reference genes for normalizing expression in cucumber according to NormFinder analysis.

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Table 5.

Expression stability of five reference genes in cucumber analyzed using the BestKeeper algorithm.

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Table 6.

RefFinder ranking of seven reference genes in cucumber in descending order of expression stability from 1 to 7.

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Fig 3.

Expression profile of miR159 and validation of selected reference genes in different cucumber tissues infected with CGMMV.

Error bars represent the mean of three technical replicates ± SD.

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