Dissertation > Excellent graduate degree dissertation topics show

Cloning of 42kDa Chitinase Encoding Gene from Trichoderma Viride LTR-2 and Construction of Transgenic Tomato

Author: SunHongXing
Tutor: YangHeTong；HuangYuJie
School: Shandong University of Technology
Course: Biochemistry and Molecular Biology
Keywords: Trichoderma Chitinase Leaf disk Tomato Plant expression vector Southern hybridization
CLC: Q943.2
Type: Master's thesis
Year: 2008
Downloads: 175
Quote: 0
Read: Download Dissertation

Abstract


Tomato as a major economic crops and genetics of model organisms has important research value, and is widely used in plant genetic engineering research. Through to insert exogenous resistance gene, construction of transgenic disease-resistant tomato, is an effective way to reduce the use of pesticides and cultivating a new crop varieties. The chitinase (Chitinase) is a class of cell wall degrading enzymes commonly found in a variety of animals, plants and microorganisms. Chitinase encoding gene transgenic plants built effective against a variety of pathogens. Different sources, however, the chitinase antiviral effect very different, so found a suitable chitinase encoding gene is of great significance for construction of transgenic disease-resistant plants. Trichoderma as important biocontrol fungi, heavy parasitic chitinase able to degrade the cell wall of a variety of pathogenic microorganisms. The study confirmed that Trichoderma produce a variety of chitinase, 42kDa chitinase degradation best kind of pathogen cell wall. 42kDa chitinase encoding genes in Trichoderma in integrated into the plant chromosome resistant plants is one of the currently constructed transgenic plant research. Trichoderma viride LTR-2 (Trichoderma virede) from vegetable plant roots in soil by the laboratory screening, its preparation has gained national invention patents, and has been registered in the Ministry of Agriculture as a new pesticide, on a variety of pathogens has good antagonism, its mechanism of action is one of a variety of cell wall degrading enzymes, especially 42kDa chitinase. From Trichoderma viride LTR-2 genomic DNA amplification by PCR technology already on the GenBank 42kDa chitinase enzyme coding gene sequence design appropriate primers to a sequence, sequencing results showed that the size of the encoding gene fragment of 1508bp , including an open reading frame of 1459bp start codon is located 45bp, stop codon located 1501bp encoding amino acid 424. Through GenBank sequence alignment, the sequence with other Trichoderma Viride 42kDa chitinase amino acid sequence having 99% homology, and thus determine the sequence of the 42kDa chitinase gene encoding the sequence in GenBank on registration (GenbankID: EF635427). Connected to the a 42kDa chitinase genes with plasmid pCAMBIA1300 CaMV35S promoter and 35S-polyA terminator, inserted into a plant expression vector pCAMBIA1302 multicloning site, to construct plant expression on carrier pCHI1302-42, using the freeze-thaw method of the carrier pCHI1302-42 is introduced into Agrobacterium tumefaciens LBA4404. Leaf disc transformation method using Agrobacterium-mediated the 42kDa gene into the tomato tissue culture regenerated plants of tomato. Finally, using PCR and Southern blot renewable Transformation of Tomato detection proved 42kDa chitinase gene has been inserted into tomato chromosome genome.

Related Dissertations

  1. Mutagenesis of the Serratia marcescens chitinase,TQ925
  2. Construction of egfp Marked-Mutants from Pseudomonas Syringae Pv.tomato and Pv.tabaci,S436.412
  3. Function of Chitinase in Lysobacter Enzymogenes OH11,TQ925
  4. Cucumber Growth Promotion and Wilt Disease Suppression of Bacillus Subtilis SQR9 and Their Mechanisms,S436.421
  5. Tolerance of Tomato on Rare Earth Elements and Its Remediation Potential on Polluted Environment,S641.2
  6. Studies on Changes of Quality Characters and Prediction Model of Postharvest Tomato Fruit,S641.2
  7. Utilization of Echinocandin B Fermentation Waste,X787
  8. Effects of Different Rootstock Grafting on Yield, Quality and Seedling Calcium Chloride Tolerance in Tomatoes,S641.2
  9. Study on Growth and Physiological Metabolism of Tomato Seedlings under High Temperature Stress,S641.2
  10. Heme Oxygenase is Involved in Salinity-Induced Tomato Lateral Root Development and Its Relationship with Hydrogen Peroxide,S641.2
  11. Molecular and Physiological Regulation Mechanism of the Nitrogen and Organic Acid Metabolism in Tomato Fruit by Different Nitrogen Forms,S641.2
  12. QTL Analysis of Salt Tolerance from Solanum Cheesmanii during Seed Germination and Seedling Stages Using Advanced Backcross Population,S641.2
  13. The Effect of Reducing Fertilization on Yield and Quality of Greenhouse Tomato,S641.2
  14. Effect of Light Environment Control on the Growth and Development in Plant,Q945
  15. Purification and Some Properties of α-Amylase Inhibitor from Avena Nuda,S512.6
  16. Study on Effects of Exogenous Auxin on Fructose Metabolism in Tomato Fruits,S641.2
  17. The Involvement of Heme Oxygenase/Carbon Monoxide System in Cobalt Chloride-Induced Lateral Root Development in Tomato Seedlings,S641.2
  18. Studies on Stress Resistance, cDNA Microarray and Carbohydrate Metabolism in Tomato Seedlings under Low Night Temperature,S641.2
  19. Digital Simulation of Temperature in Modern Greenhouse,S625.51
  20. Research and Development of Vegetable Chips,TS255.52
  21. Preliminary Study of Tomato Transformation with Soybean Genome DNA by Low Energy Ion Beam Implantation,S641.2

CLC: > Biological Sciences > Botany > Plant Cell Genetics > Plant Genetic Engineering
© 2012 www.DissertationTopic.Net  Mobile