Abstract
Lilium is an important commercial market flower bulb. qRT-PCR is an extremely important technique to track gene expression levels. The requirement of suitable reference genes for normalization has become increasingly significant and exigent. The expression of internal control genes in living organisms varies considerably under different experimental conditions. For economically important Lilium, only a limited number of reference genes applied in qRT-PCR have been reported to date. In this study, the expression stability of 12 candidate genes including α-TUB, β-TUB, ACT, eIF, GAPDH, UBQ, UBC, 18S, 60S, AP4, FP, and RH2, in a diverse set of 29 samples representing different developmental processes, three stress treatments (cold, heat, and salt) and different organs, has been evaluated. For different organs, the combination of ACT, GAPDH, and UBQ is appropriate whereas ACT together with AP4, or ACT along with GAPDH is suitable for normalization of leaves and scales at different developmental stages, respectively. In leaves, scales and roots under stress treatments, FP, ACT and AP4, respectively showed the most stable expression. This study provides a guide for the selection of a reference gene under different experimental conditions, and will benefit future research on more accurate gene expression studies in a wide variety of Lilium genotypes.
Highlights
The genus Lilium is one of the most valuable commercial market flower bulbs in the world, mainly owing to its ornamental function as a cut flower or as a potted plant
The specificity of the amplicons was confirmed by the presence of a single peak in melting curve analyses following quantitative real time PCR (qRT-PCR) (S1 Fig), and no products were detected in negative controls
It has often been assumed that the choice of stably expressed reference genes for normalization is paramount to accurate interpretation of the results [12,44]
Summary
The genus Lilium is one of the most valuable commercial market flower bulbs in the world, mainly owing to its ornamental function as a cut flower or as a potted plant. Many Lilium species and cultivars are valued for their magnificent and showy flowers, more or less recurved tepals, distinctive fragrance, wide adaptability to soils and climates, and resistance to biotic stresses [1,2,3,4] These characteristics have encouraged widespread biochemical, physiological and molecular biological studies of Lilium [5,6,7,8]. Unicolor from China, is famous for its economic and ornamental value resulting from its deep red and reflexed petals It has long been thought of as the best edible lily in China since the scales are jade white and thick, glutinous and sweet, delicate, and without residue. Since genomic resources for Lilium are still scarce, the analysis of Lilium genes, gene transcription and expression has been slow since most of the genes remain unknown
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