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本研究选取茅苍术转录组数据库中18S rRNA、β-Actin、EF-1α、GAPDH、UBQ1和UBQ2作为候选内参基因,运用实时荧光定量PCR技术分析6种候选内参基因在正常和干旱胁迫条件下不同组织中的表达,并用Delta CT、geNorm、Norm-Finder、Best Keeper及RefFinder等方法评价候选内参基因的表达稳定性。结果表明,UBQ2和EF-1α在正常条件下不同组织中的表达均较稳定,在干旱胁迫下EF-1α的表达最稳定。在此基础上,以UBQ2作为内参基因,研究正常条件下茅苍术活性成分生物合成关键酶HMGR、FPPS基因在其开花初期不同组织中的表达特性,结果显示花中HMGR、FPPS基因相对表达量均最高,根茎次之,茎和叶中相对表达量均较低。
In this study, 18S rRNA, β-Actin, EF-1α, GAPDH, UBQ1 and UBQ2 were selected as candidate reference genes in the Atractylodes lancea transcriptome database, and the six candidate reference genes were analyzed under real and drought stress conditions by real-time fluorescence quantitative PCR The expression of candidate internal reference genes was evaluated by Delta CT, geNorm, Norm-Finder, Best Keeper and RefFinder. The results showed that the expression of UBQ2 and EF-1α in different tissues under normal conditions were stable, and the expression of EF-1α was the most stable under drought stress. On this basis, using UBQ2 as a reference gene to study the expression characteristics of HMGR and FPPS, a key enzyme involved in the biosynthesis of key enzymes in the active ingredients of the Atractylodes lancea under normal conditions, the relative expression levels of HMGR and FPPS The highest, followed by the rhizome, the relative expression amount in stem and leaf was lower.