Rice Os-mir396 Overexpression Transgenic Plants To Obtain And Possible Functional Analysis

MicroRNAs play very important roles in the development of plants and animals. At present, the researchs of revealing the function of microRNA mainly focused on Arabidopsis. In the present study,we did exploratory research on the function of the 10 microRNA of rice. The target genes of these 10 microRNAs are F-box proteins, GRF(Growth regulational factor) and unknown proteins, and the anticipative functions are affecting flowering time or length of the caudex, signal transduction and stress tolerance. The main results obtained are as follows:1. Successfully amplified the precursor DNA of the 10 microRNAs from rice genomic DNA through devising primer based on the structure of the microRNA's precursor and insert it under the Ubiquitin I constitutive promoter.2. Constructed the 10 over-expressing vector and transferred them into rice callus using agrobacterium tumefaciens.After several steps of screening, we successfully obtained the T0 rice plantlet.3. By the morphological study, the over-expressing rice plantlets of T₀ generation have obvious dwarf trait, the height of them is only a half of that of the wild type. The seeds of T₁ generation bourgeoned about 5-7days later than the wild type, then presented growth retardation and dwarf trait in the seedling stage.4. Do the PCR staining on the genomic DNA of the transgenetic plants, confirm they contain the aim gene fragment.