Screening of Reference Genes for RT-qPCR Analysis of Anthocyanin Biosynthesis Related Genes in Cauliflower Curd
When cauliflower is harvested,the curd typically becomes purple,which is strongly related to the an-thocyanin content.To date,no reference genes for evaluating the expression of related genes in the anthocyanin production pathway in cauliflower have been discovered.White curd tissue in the none-turning purple curd,white curd,light purple curd and dark purple tissue in turning purple curd were employed as research materials in this study.The expression of 6 potential reference genes in various colored cauliflower curd was determined using RT-qPCR.The ΔCt method,GeNorm,NormFinder,Bestkeeper,and RefFinder software were used to assess the stability of potential reference genes.The stability of six putative reference genes in cauliflower was determined to be SKIP16＞ACT＞PP2A＞TUB＞UBQ＞EF1α.SKIP16 ranks in first in terms of overall stability,followed by ACT.They are suitable reference gene candidate for researching genes involved in the biosynthesis of anthocyanins in cauliflower.By calculating the paired variation value with GeNorm software,the best number of candidate refer-ence gene pairs was 2,and the SKIP16 and A CT genes were fully picked.The findings of this study can be utilized as a guide for accurately determining associated genes in the cauliflower anthocyanin synthesis pathway.
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