| Maize ear development dysplasia will result in an empty bar, strong adverse phenomenon, such as serious effects on maize yield. Previous research of maize ear focused on the aspects such as the growth of maize seeds after pollination. The sthdy of maize ear in aspects of mi RNAs regulation from early development is less. A new technology of degradome sequencing i was used to find maize ear development related mi RNAs and target genes. The analysis to identify the the key genes in the process of ear development. The result may make a foundation for cloning and research of ear development for in further.Micro RNAs(mi RNAs) is a kind of endogenous small molecule RNA, with the length of 20-24 nt.The non-coding mi RNA is highly conserved in various animal, plant and microbial species. The plant mi RNAs act at the transcriptional and post transcriptional level, and play an important role in plant growth and development.Most of the previous studies focused on the development of the maize kernel after pollination, and the research on the ear from the growth cone formation to the floral organ formation stage was less.The parental of a good Chinese maize hybrid seed Zheng dan958, Chang 7-2 and Zheng 58 was used as experimental material. We obtain the maize ear development related mi RNAs and their target genes, using degradation sequencing technology.The expression of mi RNA and its target genes were verified by real-time fluorescence quantitative PCR(Real-time PCR). The main results were as follows:1.We got A total of 175214241 raw readsby Illumina sequencing. Each sample average reads was about 19468249. In addition to sequencing reads, the adapter sequence and sequence shorter than 18 nt, the reads was 168052273, and each sample average net reads was 18672474, ranging from 16909865 to 22244845. Except all other types of small RNA, there are a total of 138600762 candidate mi RNA reads, each sample 15400084 reads, ranging from 11902891 to 19538721. The candidate mi RNA was compared with the corn B73 genome(Ref Gen_v2), and the 21 nt mi RNA was the most abundant in all the samples, accounting for 61.60% of all. The nexthighest reads fragment was 19 nt, representing a total of 15.51%, which represents the typical length of the mi RNA representing the mature mi RNA of the maize.2. 16 mi RNA families, 95 mi RNAs corresponding 47 target genes were found from the results of target genes accurate prediction, with mi RNA targetfinder and m RNA sequences pairing predictions and damage density file.3. 47 target genes were Gene Ontology(go) classified. The nucleus and CCAAT binding factor complex target genes accounted for 78.5%; target gene involved in transcription, DNA dependent, transcription regulation, cell differentiation accounted for 53.2%; the target gene related to DNA binding and sequence specific DNA binding, metal ion DNA binding to accounted for 72.9%.4.Expression profile of selected seven conserved mi RNAs and their corresponding target genes wasconstructed by fluorescence quantitative PCR method. The results consistent with sequencing result, indicating the reliability of the sequencing results.5.According to previous related mi RNA research reports, combined with results of this study.The analysis of differentially expressed mi RNA, indicated that the maize ear developmental processes are dependent metabolic pathway of mi RNA regulation. |
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