Selection and Validation of Reliable Reference Genes for <i>Liquidambar formosana</i> Leaves with Different Leaf Colors
2024
Fangwei Zhou | Liang Xu | Congguang Shi | Shaozong Yang | Yahui Chen
<i>Liquidambar formosana</i> Hance is renowned for its rich leaf color and possesses notable advantages, such as robust adaptability, strong resistance to diseases and pests, and rapid growth, making it a preferred choice for urban greening and carbon sequestration forest initiatives. The completion of whole-genome sequencing of <i>L. formosana</i> has spurred an increased interest in exploring the molecular mechanisms underlying seasonal changes in leaf color, marking a significant focus in <i>L. formosana</i> breeding research. However, there is currently a lack of stable reference genes suitable for analyzing the expression patterns of functional genes in <i>L. formosana</i> exhibiting varying leaf colors. This study selected five <i>L. formosana</i> varieties with significant differences in leaf colors. Through the RT-qPCR analysis, and evaluation using BestKeeper, geNorm, NormFinder, Delta Ct, and RefFinder, the expression stability of 14 candidate reference genes was examined. Consequently, two reference genes (<i>LifEF1-α</i> and <i>LifACT</i>) with stable expression, suitable for RT-qPCR of <i>L. formosana</i> with diverse leaf colors, were identified. The stability of these selected reference genes was further validated by examining the <i>LifbHLH137</i> gene, which promoted the biosynthesis of anthocyanins. This advancement facilitated molecular biology and genetic breeding investigations of <i>L. formosana</i>, providing essential data for the precise quantification of functional genes associated with leaf color variation.
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