Screening And Validation Of Tiller Related MiRNA In Barley

Barley(Hordeum vulgare L)is one of the most important cereal crops in the world.Tiller is one component of yield for barley and controlled by miRNA.In order to identify tiller-related miRNAs in barley,low-tillering mutant GSHO1990 and wild type Bowman in this study were used as experimental materials based on high-throughput sequencing and bioinformatics prediction.The main results were as follows:1.From first leaf unfolded stage to five leaf unfolded stage,in total 1608 miRNAs were identified in two barley materials.And 505 known miRNAs were identified after removing redundancy and comparing to miRBase database.207 novel miRNAs were identified under miRDeep and psRobot software.2.To set Log|FC|>1 and FDR<0.01 as conditions,a total of 15 miRNAs significantly expressed at different developmental stages and the expression trend was unanimous.And according to the function of target genes,in total 37 miRNAs were selected as candidate miRNAs.Among the 52 miRNAs,the function of target gene of chr4H:400959067..400959145:+(miR400H)was predicted to be tiller-related.The expression trend of miR400 H was unanimous and determined as a novel miRNA in barley.So it was determined for functional verification.3.To set Log|FC|<0.5 as screening condition,in total 13 candidate genes were selected for screening miRNA reference genes for barley tiller.Under three software geNorm、Normfinder and Bestkeeper and to be verifird by two tillering related miRNAs,the most stable genes in barley tiller are miR5051 and miR5048a;4.The vector construction of miR400 H was used to transfer to rice and obtained 23 transgenic plant.Additionally,a total of 100 immature barley embryos were infected and green bud has already induced from tissue culture.