ABSTRACT

Amaranth is rich in betalain. Both betalain metabolites and plant growth are affected by external factors. However, studies of amaranth seedling growth and betalain metabolism are limited by a lack of appropriate reference genes for qRT-PCR analyses. We selected 13 candidate reference genes from the amaranth transcriptome database and tested their expression stability under various environmental conditions and plant growth regulators. Five methods were used for the analysis: geNorm, NormFinder, BestKeeper, ΔCt, and RefFinder. The most stable reference genes were SAND and RAN1 for light quality, SAND and EIF5a for photoperiod, Actin-7 and TuA for temperature, SAND and Actin-7 for different tissue, SAND and EIF5a for NaCl, SAND and 60S RPL for GA₃, 28S rRNA and 18S rRNA for 6-BA, 18S rRNA and 60S RPL for 2,4-D, EIF5a and 28S rRNA for ABA, Actin-11 and SKIP35 for SA, 28S rRNA and Actin-7 for MeJA, SAND and EIF5a for all samples. To further verify the reliability of the reference genes identified in this study, the expression levels of CYP76AD1 were studied in amaranth. This is the first systematic analysis aimed at the selection of reference genes for qRT-PCR in A. tricolor under different culture conditions.

摘要

苋菜富含甜菜素。甜菜碱代谢物和植物生长都受到外部因素的影响。然而，由于缺乏用于 qRT-PCR 分析的合适参考基因，苋菜幼苗生长和甜菜碱代谢的研究受到限制。我们从苋菜转录组数据库中选择了 13 个候选参考基因，并测试了它们在各种环境条件和植物生长调节剂下的表达稳定性。分析使用了五种方法：geNorm、NormFinder、BestKeeper、ΔCt 和 RefFinder。最稳定的参考基因是SAND和RAN1为质量轻，SAND和eIF5A的用于光周期，肌动蛋白7和图阿温度，SAND和肌动蛋白7对于不同的组织，SAND和eIF5A的为氯化钠，SAND和60S RPL为GA ₃，28S rRNA的和18S rRNA的6-BA，18S rRNA基因和60S RPL 2,4- d，的eIF5A和28S rRNA的对ABA 、肌动蛋白11和SKIP35用于SA，28S rRNA和肌动蛋白7用于MeJA、SAND和EIF5a用于所有样品。为了进一步验证本研究中鉴定的参考基因的可靠性，在苋菜中研究了CYP76AD1。这是第一次系统分析，旨在选择不同培养条件下三色曲霉 qRT-PCR 的参考基因。