Dissertation > Excellent graduate degree dissertation topics show

Detection of Anti-TMV Activity of Brucein D by Green Fluorescent Protein Labeling

Author: DongLei
Tutor: ChenQiJian
School: Fujian Agriculture and Forestry University
Course: Plant Pathology
Keywords: Tobacco mosaic virus Active substances Green Fluorescent Protein Reporter gene Benthamiana Viral vectors Protoplast
CLC: S435.72
Type: Master's thesis
Year: 2010
Downloads: 38
Quote: 1
Read: Download Dissertation

Abstract


Tobacco mosaic virus (Tobacco mosaic virus, TMV) is one of the most serious virus infecting tobacco plants, and the wide distribution of the disease, resulting in huge economic losses. For these diseases, the prevention and control of active substances of plant origin is an effective and environmentally friendly measures. Therefore, screened out from the plant have a very good inhibitory effect on the virus active substance is very important. Of the laboratory has been from Chinese herbs Brucea Javanica (Brucea javanica (L.) Merr) in isolated an active substance - the Bruceine D (Brucein D), after a preliminary determination of tobacco mosaic virus (TMV) good inhibition. The expression of the green fluorescent protein (green fluoresence protein, GFP) as a reporter gene can be more intuitive and convenient observation of TMV in the receptor plants. In this experiment, the GFP gene is fused to the TMV gene constructed vector p35S: 30B: GFP for pilot viral vectors, respectively, from a study of the level of tobacco plants and protoplasts level the plant source active substance Brucein D inhibitory effect on plant viruses of TMV. By agroinoculation TMV containing the GFP reporter gene expression vector p35S: 30B: GFP injected into benthamiana leaves mixed with different concentrations of Brucein D processing injection, after a period of time by observing the expression of green fluorescent protein , screened Brucein D inhibit TMV best work concentration 10μg/mL; newborn leaves two weeks after the injection, the s tobacco PCR and western blot detection, in not add Brucein D blades to CP detected GFP gene and CP, GFP protein expression; added 10μg/mL Brucein D leaves not detected on these two genes and expression of the two proteins. The laser scanning confocal microscope in not added Brucein D blade observed containing a green fluorescent cells; 10μg/mL Brucein D leaves no green fluorescence was observed, consistent with the former results. The results also found Brucein D inhibition of TMV timeliness benthamiana plants by observing Agrobacterium transfection on the expression of green fluorescent protein to determine its inhibition of proliferation of effective time of 3-4 weeks. The present's isolated tobacco mesophyll protoplasts, protoplasts isolated yield several factors affecting the results show that when the hydrolysis time was 4 hours, the site of enzymatic hydrolysis leaves collected from plants upper newborn just commenced vacuum filtration 30 min, the enzymatic solution pH of about 6.0, the number of isolated protoplasts most for the best combination. Using PEG-mediated method successfully be transformed with foreign DNA to the present's tobacco protoplasts, detected after 48 hours, reporter gene GFP expression. Protoplast suspension plus of 10μg/ml Brucein D and without Brucein D for different processing, the results also showed Brucein D can inhibit TMV in vitro cell proliferation. Using friction inoculated with the TMV virus particles inoculated into ordinary tobacco K326 strain, two weeks after the ELISA test, the results show that, Brucein D also can effectively inhibit the proliferation of TMV in vivo in an ordinary tobacco.

Related Dissertations

  1. Breeding of 1, 3-propanediol-overproducing Strain by Genome Shuffling,TQ923
  2. Syntheses of Novelbenzopyran Compounds Containing Fluorine and Induced Resistance to TMV,S435.72
  3. The Construction and Evaluation of a Novel Non-viral Gene Transfection System and Its Application in Mesenchymal Stem Cells Gene Recombination,R346
  4. Promoter Activity Analysis of Cytochrome P450 Gene CYP9A17v2 from Helicoverpa Armigera (H(?)bner),S435.622
  5. Identification of Genes Related to Virulence of Fusarium Graminearum and Biopaning of DON Toxin Specific Affinity Peptides,S432.4
  6. Construction and Application of the Transgenic Yeast Strain Expressing Metallothionein,Q78
  7. Breeding of High-Yield Nosiheptide Strain and Studying of Its Formation Condition,TQ936.16
  8. Studies on Flocculating Mechanism of Lactobacillus Flocculating Starch in Sweet Potato Slurry and Isolation, Identification and Characterization about Active Ingredients,TS231
  9. The Toxic Threshold by Biuret to Crops in the Compound Fertilizer and Research on the Degrading Bacterium of Biuret,S143
  10. PPARs established based on human cells that target drug screening platform,R965.1
  11. Studies on Nano-drug Delivery System of Co-delivery of Gene and Doxorubicin,R943
  12. Construction of Plant Expression Vectors with Multi-Selectable Marker,Q943.2
  13. Anti-tobacco mosaic virus drug screening and preliminary study on the mechanism of,S482.2
  14. Dissertation for Master’s Degree Fujian Agriculture and Forestry University,S646
  15. The Research on the Optimized Transformation System of the Green Fluorescent Protein Gene to Fusarium Oxysporum from Solanaceae Crop and Its’ Colonization Characteristics,S436.41
  16. Construction and Genetic Transofrmation of the Rice Chloroplast-specific Expression Vector,S511
  17. Glucagon - like peptide-1 and its analogs gene therapy for diabetes research,R587.1
  18. Polymorphism of Serotonin Transporter Gene in the Promoter Region and Irritable Bowel Syndrome,R574
  19. Cloning and Function Identification of the Promoter of Δ6-Fatty Acid Desaturase Gene from Mucor sp.,Q78
  20. Construction of Genetically Engineered Streptomyces,Q78
  21. Study on Preparation and Application in Treatment of Petrochemical Wastewater of Microbial Flocculant,X703.5

CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Crop pests and diseases and their prevention > Economic crop pests and diseases > Tobacco pests and diseases
© 2012 www.DissertationTopic.Net  Mobile