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Transformation of Oat with PMSAMS from Broomcorn Millet and Cloning of a DREB Gene from Foxtail Milletand Its Expression during Drought Stress

Author: YangXiWen
Tutor: HuYinGang
School: Northwest University of Science and Technology
Course: Crop Genetics and Breeding
Keywords: Oat Millet Drought-Tolerant Related Genes Genetic Transformation RT-PCR
CLC: S515
Type: Master's thesis
Year: 2011
Downloads: 66
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Abstract


Drought occurs frequently and becomes the bottleneck of agricultural development in many areas. Crop genetic improvement on drought resistance is so urgent to overcome drought stress. The selection of drought genes and the use of drought-related genes have become an important prerequisite for breeding of drought- tolerance variety. In this study, plant expression vector with PmSAMS gene from broomcorn millet was constructed and used to transform oat with Agrobacterium by in-planta method and floral dip method. A DREB gene was cloned from foxtail millet by RT-PCR, and its expression patterns during drought stress were investigated by means of semi-quantitative RT-PCR. The main results were as follows:1. The fulllength cDNA of osmotic stress-related gene PmSAMS, was amplified from broomcorn millet, which with the 3 typical domains of SAMS (N terminal domain, inter domain and C terminal domain), and inserted into plant expression vector pCAMBIA1305. In which, the PmSAMS gene was driven by promoter CaMV35S.2. The PmSAMS gene which were driven by promoter CaMV35S was used to transform oat with in-planta method and floral-dip method by Agrobacterium mediated method. Concentration of Hygromycin (500mg/L) was dertermined in oat for screening of transformants. One resistant seedling was obtained and confirmed by PCR.3. A DREB gene was amplified from Foxtail millet which was 1 113 bp in length, encoding 259 amino acids with basic amino acid regions possible nuclear location sequences(NLSs) and a highly conserved AP2/EREBP domain in the encoded putative protein. The expression patterns of DREB from Foxtail millet during drought stress were investigated by means of semi-quantitative RT-PCR. The results showed that DREB from Foxtail millet might be one of the key genes for drought tolerance and water use efficiency of foxtail millet.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Millet ( millet, millet)
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