Table 1.
Details of primers used for qRT-PCR analysis.
Table 2.
Description of different tissue samples used for qRT-PCR analysis.
Fig 1.
Ct variation and expression stability analysis of each candidate reference gene among different tissue samples using geNorm.
(a) Boxplot depicting absolute Ct values, which was calculated using GenEx program. Lower and upper boxes indicate the 25th and 75th percentile, respectively. The median is depicted by the line and all outliers are indicated by dots (b) Gene expression stability graph based on average expression stability values (M-value), using stepwise exclusion process. The lower the M-value indicates, higher the stability of gene. The direction of the arrow indicates the most and least stable housekeeping genes.
Fig 2.
Ct variation and expression stability analysis of each candidate reference gene among different tissue samples using NormFinder.
(a) Boxplot depicting absolute Ct values, which was calculated using GenEx program. Lower and upper boxes indicate the 25th and 75th percentile, respectively. The median is depicted by the line and all outliers are indicated by dots. (b) Gene expression stability graph using NormFinder algorithm based on stability values. Lower the stability value indicates higher stability of the housekeeping gene. The direction of the arrow indicates the most and least stable housekeeping genes.
Table 3.
Ranking of tested housekeeping genes for drought stress conditions using geNorm and NormFinder algorithms.
Fig 3.
Validation of selected housekeeping genes under drought stress conditions.
Expression profiling of candidate gene responsible for universal stress protein A-like (uspA) protein in drought imposed tissues (root, stem and leaves). The expression value of candidate gene was normalized with stable, combination of stable and least stable genes namely (i) IF4α (ii) TUB6 (iii) HSP90 (iv) IF4α + TUB6 (v) IF4α + HSP90 (vi) IF4α + TUB6+HSP90 (vii) ACT1 and (viii) 18SrRNA. The relative quantitative values of selected drought responsive candidate gene were obtained after scaling to control samples. EDR: vegetative root stressed; LDR: reproductive root stressed; EDS: vegetative stem stressed; LDS: reproductive stem stressed; EDL: vegetative leaves stressed; LDL: reproductive leaves stressed.