Selection and validation of reference genes for quantitative gene expression studies in Erythroxylum coca

Area of Science:

• Molecular Biology
• Plant Science
• Biochemistry

Background:

• Real-time quantitative PCR (qPCR) is vital for comparative gene expression studies.
• Accurate qPCR relies on stable reference genes for normalization.
• Erythroxylum coca is a model for cocaine biosynthesis, requiring reliable gene expression analysis.

Purpose of the Study:

• To identify stable internal reference genes for Erythroxylum coca.
• To enable accurate comparative gene expression analysis in coca tissues.
• To support the development of E. coca as a model species for alkaloid biosynthesis research.

Main Methods:

• Evaluated expression stability of nine candidate reference genes in E. coca.
• Measured gene expression across seven tissues (flowers, stems, roots, four leaf stages).
• Assessed stability using geNorm, NormFinder, and BestKeeper algorithms.

Main Results:

• Ec10131 and TPB1 demonstrated high expression stability in coca leaves.
• Ec10131 and Ec6409 were identified as the most stable reference genes across all tested tissues.
• Expression stability varied significantly among candidate genes and tissues.

Conclusions:

• Ec10131 and TPB1 are recommended for reference gene selection in coca leaves.
• Ec10131 and Ec6409 are suitable for normalization in broader E. coca tissue studies.
• Validated reference genes are essential for advancing research on coca plant gene expression and biosynthesis.