solation of cleaved RNA fragments is laborious, time consuming and expensive. Recently, high throughput sequencing methods, known as degradome analysis or PARE that can globally EPZ-5676 leukemia identify small RNA targets have been developed to overcome such limitations. Soybean is one of the most important crops cultivated all over the world. It is a good source of vegetable pro tein and oil. However, the role of miRNAs in soybean seed development is mostly unknown. So it is important to identify the seed developmental stage specific and tissues specific miRNAs and their potential target genes. Identification of the consequences of miRNA guided tar get degradation that occurs in a developmental and tis sue specific manner could help to elucidate how lipid and protein metabolic pathways operated during seed development.

The soybean genome was decoded a year ago, and this information has accel erated molecular research on soybeans. Although many soybean miRNAs were identified in previous research, the number of miRNAs known in soybean is still very small and considerably lower than that in Ara bidopsis or rice. High throughput sequencing technolo gies such as massively parallel signature sequencing, 454 and sequencing by synthesis have enabled the identification of miRNAs in soybean. The extent of miRNA directed post transcriptional gene regulation in any organism can only be fully realized by identifying not only the miRNA component but also the set of their RNA targets.

Recently, miRNA targets have been reported for one of the many stages of soybean seed development, namely very early at 15 days after flowering, and without dissec tion of the maternal seed coats from the cotyledons which develop from the zygote. To comprehensively investigate small RNA targets and provide basic infor mation for further understanding of the miRNA mediated post transcriptional regulation during different soybean seed developmental stages, we constructed five separate degradome libraries derived from seed coats and cotyledons of different developmental stages repre senting the early, mid, and late maturation stages of seed development. The libraries were sequenced using SBS sequencing technology. The degradome dataset for the five different libraries was computationally analyzed. The majority of these reads mapped to the soybean transcrip tome.

A total of 183 target genes were confirmed as miRNA targets, which included both conserved and non conserved miRNAs. Additionally, we have identified Anacetrapib targets for 25 cotyledon neither specific miRNAs, as well as 12 miRNAs and their potential targets found only in the seed coats. We found 16 miRNA families and their large number of targets that are found in both tissues. More over, we have validated Auxin Response Factors to be targets of gma miR160, as verified by RNA ligase mediated 5 rapid amplification of cDNA ends. The identification of developmental stage specific and tissue specific miRNA targets including many transcription factors advance ou