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Cloning and Expression Analysis of VpUSP Gen in Chinese Wild Vitis Pseudoreticulata

Author: ZhaoZuo
Tutor: WangYueJin
School: Northwest University of Science and Technology
Course: Of Horticultural Plant Germplasm Resources
Keywords: Chinese wild grapes RACE Grape powdery mildew
CLC: S663.1
Type: Master's thesis
Year: 2010
Downloads: 76
Quote: 0
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Abstract

Stress protein (Universal stress protein, USP) is a widespread resistance related protein in plants, is generally present in the cytoplasm by the upregulation of external stress in plants, enhanced stress resistance in plants. Grape powdery mildew [Uncinula necator (Schw.) Burr.] Seriously endanger the world grape production fungal diseases. In the grape famous producing areas such as California, France, Italy, Australia, China, Xinjiang and other regions, grape powdery mildew damage is more serious, causing huge economic losses to grape production. The combination of the use of genetic engineering and traditional breeding techniques, discover resistance genes for disease-resistant grape germplasm to breed new varieties of disease-resistant is one of the effective ways to solve this problem. Therefore, the usp gene to carry out China's Wild East grapes molecular biology research has a very important significance. To grape strains according to the Chinese wild grapes from our group pre-build Eastern Shirakawa -35-1 a powdery mildew induced cDNA library with a wide range of stress protein related EST sequences, primers were designed, and RACE clone Chinese wild the Huadong grapes Shirakawa -35-1 stress protein gene (Universal stress protein, USP) full-length sequences for sequence analysis by semi-quantitative RT-PCR technology, research VpUSP Wild East China grape disease resistance, sense of diseased plants Department of expression changes. The main findings are as follows: (1) and RACE amplification in Chinese wild the Huadong grapes Shirakawa-35-1usp the gene (VpUSP), bioinformatics analysis showed that the gene is 836 bp open reading frame of 528bp, encoding 175 amino acids , GenBank accession number GU946975. Chinese wild the Huadong grapes Shirakawa -35-1 VpUSP gene deduced amino acid sequence for the retrieval sequence through NCBI BLASTx program detects their amino acid sequence homology was found with similar sequences: Arabidopsis thaliana, beans, rice, homologous is between 59% -62%, showed that the sequence was obtained for the USP gene family members. (2) the gene expression patterns induced by the powdery mildew: stress proteins in the Wild East grapes sense of diseased Department powdery mildew before and after vaccination were expressed, but the expression levels did not change significantly; However, disease the Wild East grapes strains inoculated with powdery mildew VpUSP gene expression was significantly increasing trend and reached peak expression at 48h and then decreased, the preliminary indication of The Wild East the grapes VpUSP gene expression activity in the process of interaction with the powdery mildew fungus. (3) to Shirakawa -35-1 plantlets were used to study the combination of different hormone concentration Shirakawa -35-1 leaves and petioles, stems segment adventitious buds regeneration. The results showed that 0.6%, the induced the White River -35-1 leaves and petioles, stems segment optimum medium MS IBA 0.2 mg / L TDZ4mg / L sucrose 15g / L agar, the regeneration rate of 9%, 10%, 8%, respectively. (4) to plantlets Shirakawa -35-1 material to study the impact of combination of different hormone concentration Shirakawa -35-1 subculture, results show that best subculture medium 1/2MS IBA1.0mg / L NAA0 .1 mg / L sucrose 28g / L agar 0.6%.

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