High-Throughput Sequencing And Real Time PCR Identifacation Of MicroRNAs In Immature Maize Leaf And Root Under The NaCl Treatment

A problem in China is that the serious salinization of arable land,and the ratio of salinization in arable land continues to rise,thus solving the problem of salinization of land is desperately needed.The soil salinization cause stunted plants,the leaves turn yellow,and the growth was inhibited.In recent years,the discovery has been confirmed the miRNA with answering multiple stresses.In this experiment,maize root were split to two parts and each part was under NaCl treatment or usual treatment;small RNAs of maize’s leaves and roots were sequenced with Solexa under NaCl treatment,looking to confirm the novel miRNA and predicting the novel miRNAs’ target gene.The main results are as follows:(1)this experiment built 5 more than 3 million reads small RNA using high-throughput sequencing.Remaining 1442733-3327070 tags contain theirs information in these 5 databases separately after the further filtering.24nt sequence contains most,occupied 12%to 51%respectively in these 5 databases indicated that 24nt RNA sequences were the main type.(2)Compared sequencing result and miRBase database,the result showed there were 116-171 mature miRNAs in 30-31 families conserved in this experiment.The others sequences were predicted 234 using mireap.(3)Predicted these novel miRNAs’ target gene,184 of them have one or more targets.109 of these targeted gene cluster had clear functional annotation include function of signaling pass way,electron transport,transferase,metabolism and nucleic binding.(4)It obtained 30 novel miRNAs’ target may be related to salt stress and 19 of them were confirmed to be real by Real Time PCR.According to their target gene function and miRNAs expression level,their molecular mechanism is predicted.This Solexa sequencing result not only enrich the maize small RNA database,more importantly,the identification of miRNA and target gene prediction contributed a lot for the further study of miRNA regulation mechanism of maize under salt stress.