Identification of a dominant housekeeping genes to normalize mRNA level in dioecious Palmyra palm

Abstract

A stable housekeeping gene is required for precise and consistent qPCR data analysis. Palmyra palm is an important tropical dioecious tree crop with diverse uses. However, no reliable gene expression studies or housekeeping gene standardization have been documented, which limit crop improvement and utilization. In this work, four frequently used statistical tools such as RefFinder, NormFinder, Delta CT method and BestKeeper were applied to evaluate the constancy and fluctuation in expression of a chosen 10 internal control genes such as NADH dehydrogenase subunit 5-like (NAD5), β-actin, ACTIN CRD, alpha-tubulin, UBCE, EIF, EF2, AP1, GAPDH and UBQ2 in both vegetative and reproductive tissues of dioecious Palmyra palm. Six flower tissues and two leaf tissues from male and female palms were used in this study. The findings indicated that NAD5-2 is the most reliable and constant housekeeping gene expressed during Palmyra palm flower development. The AGAMOUS and SEEDSTICK genes expression pattern in the flower development were used to validate the normalized expression of the four housekeeping genes that were chosen in order to further support the stability of those genes. Our results will be useful for further research on gene transcription level in Palmyra palms and also the above mentioned tool is very useful for the normalization of housekeeping genes of biological samples with different experimental conditions.