Stability Evaluation of Reference Genes for Fluorescence Quantitative RT-PCR in Zizania latifolia
Zizania latifolia is a kind of perennial aquatic vegetable of Gramineae,and the edible organ was the swollen culm formed by the symbiotic of Zizania latifolia and Ustilago esculenta.The selection of suitable internal reference genes of fluorescence quantitative RT-PCR(RT-qPCR)is the basis of studying the gene expression pattern of the Z.latifolia in the interaction between host plants and fungus.In this study,roots,leaves,sheaths,stem apex and culm gall of Zizania latifolia at different stages were used as research materials.The expression levels of 11 reference genes such as β-Actin,Actin3,β-tubulin,GAPDH,UBQ5,UBC1,PK,elf1α,eIF4a,SFua2a and 18S rRNA were analyzed by RT-qPCR,and the expression stability of these reference genes were identified using the GeNorm,NormFinder,BestKeeper and RefFinder software,then the ZlADR1 gene was used for verification analysis to test the reliability of reference genes.The results showed that SFua2a and eIF4a followed by Actin3 were stably expressed in all samples and were the appropriate reference genes of Z.latifolia.
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