The Selection Of Endogenous Reference Genes In Phyllostachys Edulis Treated With Abiotic Stresses For QRT-PCR Analysis

Phyllostachys edulis belongs to Gramineae,Phyllostachys.Due to its rapid growth,excellent material quality,and wide range of uses,P.edulis has become China's bamboo species with the highest economic value.However,with the deteriorating environment,cold damage,drought,soil salinization and other factors are constantly affecting the production and quality of bamboo and bamboo shoots.Therefore,abiotic stress research has important implications for the production of bamboo.Quantitative Real-time PCR(q RT-PCR)is a commonly-used method for studying the temporal and spatial expression of genes during development currently,in which choosing appropriate reference genes is a prerequisite for accurately analyzing relative expressions of target genes.At present,there is no study of the selection of endogenous reference genes in P.edulis treated with abiotic stresses.This study took the roots,stems,and leaves of P.edulis treated with low temperature,drought and salt stresses as experimental materials and analyzed the expressions of ten kinds of candidate reference genes-TIP41,NTB,KDSA,PP2 A,SAND,e IF-4?,18 S r RNA,HSP,GAPDH and ACT in different tissues or organs by the aid of the q RT-PCR.After the comparative analysis using Ge Norm,Norm Finder and Best Keeper,the results were as fellows: 1.Under low temperature treatment,e IF-4? and TIP41 were viewed as the corrective reference genes when the genes expression in different leaves or tissues of P.edulis were analyzed and compared by q RT-PCR.2.Under salt stress treatment,when the differences of genes expression were compared in different leaves of P.edulis,e IF-4? could be chosen as the corrective reference gene.In order to compare the differences of gene expression in different tissues of P.edulis,e IF-4? or TIP41 could be chosen as the corrective reference gene.3.Under drought treatment,NTB and KDSA were the most appropriate reference genes when the differences of genes expression were compared in different leaves of P.edulis,and NTB and TIP41 were the most appropriate reference genes when the differences of genes expression were compared in different tissues of P.edulis.These findings were inconsistent with those of previous studies,which ACT was used as the only one reference gene.The results of this study would provide a theoretical basis for selecting the appropriate reference gene of P.edulis treated with abiotic stresses in the analysis of q RT-PCR.In addition,in order to explore whether the reference genes of P.edulis are suitable for other bamboo species,this study took Phyllostachys nigra,Bambusa multiplex and Sasaella kongosanensis‘ Aureostriatus ' as experimental materials and analyzed the expressions of ten candidate referencegenes of P.edulis in their different tissues or organs by the aid of the q RT-PCR.The results showed that the reference genes of P.edulis could be used for the relative quantitative PCR analysis of other bamboo species.The results of this study would provide a theoretical basis for the versatility of bamboo reference genes.