title: Identification of the valid reference genes for quantitative RT-PCR in annual ryegrass (Lolium multiflorum) under salt stress.
abstract: Annual ryegrass (Lolium multiflorum) is a cool-season annual grass cultivated worldwide for its high yield and quality. With the areas of saline soil increasing, investigation of the molecular mechanisms of annual ryegrass tolerance under salt stress has become a significant topic. qRT-PCR has been a predominant assay for determination of the gene expression, in which selecting a valid internal reference gene is a crucial step. The objective of present study was to evaluate and identify suitable reference genes for qRT-PCR in annual ryegrass under salt stress. The results calculated by RefFinder indicated that eEF1A(s) was the most stable reference gene in leaves, whereas EF1-a was the least stable; meanwhile, TBP-1 was the most optimal in roots and in all samples, and the eIF-5A shouldn't be utilized for normalization of the gene expression. eEF1A(s) is more suitable than TBP-1 as reference gene in leaves when verified with P5CS1 and Cyt-Cu/Zn SOD genes. We should choose optimal reference genes in specific tissues instead of the most stable one selected from different conditions and tissues.
authors.affiliation: Grassland Science Department, Sichuan Agriculture University, Chengdu 611130, China. wangxiayuyao@126.com.
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
authors.affiliation: Grassland Science Department, Sichuan Agriculture University, Chengdu 611130, China. maroar@126.com.
authors.forename: Xiao
authors.initials: X
authors.lastname: Ma
authors.affiliation: Grassland Science Department, Sichuan Agriculture University, Chengdu 611130, China. huanglinkai@sicau.edu.cn.
authors.forename: Linkai
authors.initials: L
authors.lastname: Huang
authors.affiliation: Grassland Science Department, Sichuan Agriculture University, Chengdu 611130, China. zhangxq@sicau.edu.cn.
authors.forename: Xinquan
authors.initials: X
authors.lastname: Zhang
mesh_terms: D020869:Gene Expression Profiling / Q000592:standards*; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D008129:Lolium / Q000235:genetics* / Q000254:growth & development; D009928:Organ Specificity; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012015:Reference Standards; D054712:Salinity; D013312:Stress, Physiological