The authors have reported Regorafenib manufacturer that miR164 directs cleavage in vivo at a position complementary to the 10th nucleotide from the 5 end of the mature sequence. The SNP found in barley is in the 11th position, therefore it is likely to prevent the clea vage and produce phenotypic effects on root development. SNPs have been identified also in other two conserved miRNA targets, TIR1 and AGO4, targeted respectively by miR393 and miR408. TIR1 is an auxin receptor nega tively regulated by miRNAs in response to bacterial fla gellin, as a defence mechanism against Pseudomonas syringae. AGO4 is a protein involved in the siRNA mediated gene silencing, and it is required for the resis tance to the same pathogen. Therefore, miR393 and miR408 are likely to work in a coupled manner during P. syringae infection.

The two SNPs identified are in the 12th position and could potentially alter the levels of pathogens resistance. SNPs were also found in previously not reported miRNA targets, such as the AWPM 19 like protein matching to the miRNA 1134. AWPM 19 accumulates in wheat plasma membrane during cold acclimation in response to abscisic acid. If this miRNA really con trols the synthesis of this protein, a deleterious SNP in the 11th position could then change resistance to cold stress. Conclusions This study has thus provided an update of the informa tion on barley miRNAs and their targets representing a foundation for future studies. Novel putative target genes have been identified and most of them are involved in stress and hormone response.

Indeed, the role of plant miRNAs in abiotic and biotic stress response as well as in auxin signalling is well known. In particular, protein kinases such as protein kinase C and serine threonine kinase, known to be important regulator on abiotic stress resistance, are largely present in novel microRNA target pairs identified. The results have also shown that microRNA target sites can be an interesting source for the identification of functional genetic variability, representing an interest ing source of candidate molecular markers for applica tion in barley breeding. Putative polymorphisms Batimastat have now to be verified by amplification and sequencing of the target sequences on a larger set of genotypes. Sequence analysis based on known miRNAs can obviously give insights only on conserved mRNAs and related targets.

Future work will thus be based on the construction of a degradome library for parallel analysis of RNA end, a powerful approach for high throughput identification vali dation of conserved and non conserved targets. Methods miRNA reference dataset The initial miRNA dataset has been obtained by extract ing the mature sequences of the Viridi plantae group from the miRBase release 13. By removing identical mature sequences, the size of this dataset has been subsequently reduced to 1014 non redundant sequences related to selleck chem Volasertib 468 miRNA families.