背景：牡丹具有显著的观赏性，药用和油的价值。脱水和盐度等渗透胁迫限制了牡丹栽培面积的扩大。关于渗透胁迫和激素刺激下牡丹参考基因选择的信息仍然有限。本研究旨在确定在渗透胁迫和激素处理下适合牡丹的稳定内参基因。为分子生物学研究提供理论依据。
结果：在渗透胁迫和激素处理下，通过RT-qPCR评估了牡丹的12个候选内参基因。DeltaCt方法,geNorm,和NormFinder用于综合表达稳定性排序比较。结果表明，微管蛋白-α是干旱和ABA处理的首选内参基因，微管蛋白β在干旱和OPDA诱导下被鉴定为最合适的内参基因，18s-rRNA被认为是盐度和JA处理最稳定的基因，eIF-5A被列为JA和MeJA治疗中最稳定的基因。实验还表明，在ABA和BR激素处理下，EF1-α相对不稳定。
结论：这些优选的内参基因可用于涉及牡丹渗透或激素应激的qPCR研究。预计该结果将有利于未来牡丹功能基因组学研究和分子育种研究。
BACKGROUND: Tree peony possess significant ornamental, medicinal and oil values. Osmotic stresses including dehydratiuon and salinity limit the expansion of cultivation area of tree peony. Information on reference genes selection under osmotic stress and hormone stimulation of tree peony still limited. This study aimed to determine the stable reference genes suitable for tree peony under osmotic stresses and hormone treatments, and provide a theoretical basis for the molecular biology research.
RESULTS: Twelve candidate reference genes were evaluated in Paeonia ostii \'Fengdan\' under osmotic stress and hormone treatments by RT-qPCR. Delta Ct method, geNorm, and NormFinder were used for the comprehensive expression stability ranking comparison. The results revealed that tubulin-α was the preferred internal reference genes for drought and ABA treatment, tubulin-β was identified as the most suitable reference gene under drought and OPDA induction, 18s-rRNA was regarded as the most stable gene for salinity and JA treatment, eIF-5 A was listed as the most stable gene for JA and MeJA treatments. The experiments also displayed that EF1-α were comparatively unstable under ABA and BR hormone treatments.
CONCLUSIONS: These preferred reference genes could be useful in qPCR studies involving osmotic or hormonal stresses in Paeonia ostii \'Fengdan\'. It is anticipated that the results will benefit tree peony functional genomics studies and molecular breeding research in the future.
