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Cloning and Functional Analysis of MicroRNAs from Populus Euphratica

Author: ZuoYuRong
Tutor: YinWeiLun；XiaXinLi
School: Beijing Forestry University
Course: Botany
Keywords: the microRNAs precursors and mature body Dehydration High salt Populus peu-MIR393a Overexpression
CLC: S792.119
Type: Master's thesis
Year: 2011
Downloads: 84
Quote: 0
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Abstract


MicroRNAs (miRNAs) are a class of about 22 nucleotide acid, small molecule non-coding RNA. miRNA can degrade or inhibit the protein synthesis of the target gene mRNA, thereby in plant abiotic stress responses play an important role. Populus (Populus euphratica) is an arid desert region in China's only natural forest of tall trees and bamboo, poplar has salinity, strong stress resistance barren, drought-resistant. Research in Abiotic Stress regulatory mechanism of miRNAs in Populus, has far-reaching value of basic research and applied research for the discovery of the mechanism of action of the miRNAs and their target proteins, cultivate new varieties of high stress resistance. In this experiment we determined by PCR amplification of 13 of the 12 family Populus 11 miRNAs miRNAs precursor and 12 miRNAs family miRNAs mature. In order to detect dehydration and high salt stress Populus of miRNAs precursor and mature expression differences, we apply SYBR Green RT-PCR method the real-time quantitative family Populus 13 miRNAs 14 miRNAs precursor and 17 miRNAs family miRNAs mature 17 body. This is the first reported the expression of the precursor and the mature form of the Populus miRNAs. Abiotic stresses stress expression by detection of of miRNAs precursor and mature body, it is possible to identify miRNA formation of the adjustment process in the face of adversity environment suffered. The majority of miRNAs predicted target genes involved in growth and development, stress resistance and metabolic processes. Populus vivo cloning experiments, we have identified five predicted target genes, and dehydration and high salt stress expression quantitative analysis of these five target genes. Based on this study, we propose a possible mechanism for poplar miRNA in abiotic stress response network. This study will contribute to our understanding of the complex regulatory networks of miRNA regulation poplar abiotic stress adaptive response. The the transgenic peu-MIR393a to the Arabidopsis (Arabidopsis thaliana) and found the the miR393a precursor transgenic plants with mature body than the wild-type Arabidopsis showed overexpression, the salt stress wild-type and T3 transgenic plants and the detection of wild-type indicators and transgenic Arabidopsis germination rate, survival rate, root length, higher than that of the control results show that the salt tolerance of the transgenic plants WRKY33 transcription factor target gene prediction and salt resistance has a negative effect, so that Populus Peu gene-MIR393a salt tolerance. In this study, the Peu-MIR393a gene specific functional studies laid the foundation for the later to carry miRNA resilience gene engineering.

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CLC: > Agricultural Sciences > Forestry > Forest tree species > Broad-leaved trees > Young > Other
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