Abstract
Eustoma grandiflorum, commonly known as prairie gentian or Texas bluebells, is among the most popular agriculturally propagated species of cut flowers. Due to its widespread appeal, there is increasing interest in understanding the molecular genetic factors underlying floral development and resistance to abiotic stresses. We analyzed 18 potential reference genes in different organs, at different floral developmental stages and under drought- and salt-stress treatments, for use in RT-qPCR analysis. A total of four analytical tool packages, including geNorm, NormFinder, BestKeeper, and RefFinder were employed to determine the most appropriate reference genes under each treatment condition. The results demonstrate that different reference genes should be used for normalization under different experimental treatments. EgPP and EgPP2A2 were the most stable internal control genes across different organ types, EgPP and Eg18S were the most stable under salt-stress, EgPP and EgACT1 were the most stable across different floral development stages, and EgEF1A and EgTUA were the most stable reference genes under drought-stress. Additional gene expression analyses of EgMIXTA1, EgTOE1, and EgP5CS1 further confirmed the applicability of these reference genes. The results represent a significant contribution to future studies of reference gene selection for the normalization of gene expression in Eustoma grandiflorum.
Highlights
Eustoma grandiflorum, commonly known as prairie gentian or Texas bluebells, are widely distributed across North America and bloom year-round [1]
The experimental samples, ‘Double Flower Development’, were collected from Eustoma grandiflorum cv. ‘620’ and others were all collected from E. grandiflorum cv. ‘2003-2-2’
Arabidopsis thaliana were used as queries to search the local transcriptome database of
Summary
Commonly known as prairie gentian or Texas bluebells, are widely distributed across North America and bloom year-round [1]. This species has become a popular, high-grade cut flower variety in the international flower market due to its long vase life, diverse colors, and multitudes of flower patterns. Research on E. grandiflorum has gradually shifted from morphological descriptions and flowering physiology to molecular genetic analyses of floral organ development, as well as tolerance to drought-, salt-, and other abiotic-stressors [2,3]. The basis of molecular genetic research, is important for understanding gene function. The accuracy of results is affected by the quality and integrity of RNA [8], the quality
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