Preliminary Studies On The Regulation Of OsMADS34 To The MicroRNA396b In Rice

Micro RNA?mi RNA?is a non-coding small molecule RNA with a length of about 21nt.It can specifically identify target m RNA and bind to it so as to degrade target m RNA and inhibit its subsequent translation and methylation.Through this series of operations,gene expression will be negatively regulated.As one of the major food crops in the world,rice production has a direct impact on global food security.With the deepening of the research on mi RNA,researchers have enhanced their understanding of the gene regulation mechanism and demonstrated a multi-level and all-dimensional regulatory system for gene expression in plant cells through its regulatory mechanism.The MADS-box family is an important transcriptional regulator involved in many aspects of rice growth and development.Depending on the protein domain,the mads-box family of genes can be divided into two types:Type I and Type II In rice,both types exist.In the previous laboratory,Henry sun constructed a high quality c DNA library with the yeast single hybridization kit,and constructed the Osmi R396b promoter bait plasmid to screen Os MADS34 and some other factors that may be related to its promoter.In this paper,the regulation effect of Os MADS34 on the promoter region of Osmi R396b was studied in depth through promoter activity detection,yeast monoheterozygous,subcellular localization and EMSA experiments.Through the study in this paper,we hope to lay a foundation for the subsequent elucidations of Os MADS34-Osmi R396b regulatory pathway,and further improve the rice yield on this basis.The specific research results are as follows:The combination of Os MADS34 and Osmi R396b promoter was confirmed by yeast mono-hybrid recovery verification.The key Domain in Os MADS34 was identified as mads-domain and then truncated to Os MADS34^88-147.Through software analysis and yeast oneo hybrid experiment,the core basis of Osmi R396b promoter region was determined to be CACCAACC.The core base was located as C through three series after the point mutation.The Osmi R396b promoter region was constructed onto a suitable vector and subcellular localization was performed by using rice protoplast transformation method.Microscopic observations showed that Os MADS34 is a transcription factor located in the nucleus.However,promoter activity detection and tobacco GUS staining experiments confirmed that the addition of Os MADS34 into the system could enhance the activity of Osmi R396b promoter.Os MADS34 was knocked out and overexpressed in vivo,and positive plants were obtained.