Dissertation > Excellent graduate degree dissertation topics show

Identification and Detection of Cucumber Green Mottle Mosaic Virus

Author: HuangJing
Tutor: WangZongHua；LuGuoDong；LinShiMing
School: Fujian Agriculture and Forestry University
Course: Plant Pathology
Keywords: Cucumber green mottle mosaic virus Mechanical inoculation DAS-ELISA IC-RT-PCR Sequence analysis
CLC: S436.421
Type: Master's thesis
Year: 2007
Downloads: 164
Quote: 1
Read: Download Dissertation

Abstract


Cucumber green mottle mosaic virus (CGMMV) is an important seed-borne virus, which is responsible for devastating losses of cucurbitaceous crops. The virus is one of the potential hazardous pests in China. At present, it has been intercepted and captured frequently by port plant quarantine department, and it has been found at several regions recently. It’s long distance spread is transmitted through international trade or domestic trade of seeds. Therefore, it is impendency to develop detection progresses of CGMMV. In this study, methods of identification and molecular detection of CGMMV were developed. These methods can help to intercept and capture the virus more efficiently. Major results are described as follows:(1) CGMMV was detected by mechanical inoculation, transmission electron microscopy, double antibody sandwich ELISA ( DAS-ELISA ) and reverse transcription polymerase chain reaction (RT-PCR) in a cucumber sample which was collected from a greenhouse in Nanning city, Guangxi province.(2) Immuno-capture RT-PCR ( IC-RT-PCR ) method was established by optimizing the RT-PCR reaction system and amplification procedures. A coat protein(CP) encoding gene was amplified by IC-RT-PCR, and the PCR product was cloned into pMD18-T vector. Sequences analysis showed that CP gene of this isolate had 486 nucleotides, encoding 161 amino acids with theoretical molecular weights of 17．3 kDa. Sequence alignment showed it shared 91．4 %～99．4 % nucleotide acid identities and 98．8 %～100 % amino acid identities with the published sequences, respectively.(3) Six pairs of primers were designed to amplify the coding region of CGMMV which were isolated from Lagenaria siceraria leaves with inoculated by single ring spot on Chenopodium amaranticolor. Six nucleotide sequence fragments were cloned, sequenced and then reconstructed. Sequence analysis showed that this isolate had 6188 nucleotides in length, encoding 1144 amino acids. Comparison showed it shared 60．1 %～98．2 % nucleotide acid identities and 58．4 %～99．0 % amino acid identities with the published sequences, respectively.(4) The IC-RT-PCR was applied directly to detect the virus infested seeds. The results showed that IC-RT-PCR could directly detect CGMMV in infected seeds, which not only simplifies the RNA extraction, but also decline the demand of samples. IC-RT-PCR is specific, simple and convenient, and could be used as a rapid and efficient detection method for CGMMV diagnosis. The IC-RT-PCR is a hopeful method in practical application.

Related Dissertations

  1. Survey in Nanjing and Analysis of the Mitochondrial Genome of Frankliniella Occidentalis (Pergande),S433
  2. Isolation and Identification of Low Pathogenicity Avian Influenza (H9) Virus in Henan and Study on the Biological Charactistics,S852.65
  3. Isolation and Identification of Infectious Bronchitis Virus and Sequence Analysis of Its S1 Gene and N Gene,S852.65
  4. Pathogenic Mechanism of Invasion Associated Genes in Avian Pathogenic E. Coli Duck Isolate DE205B,S852.61
  5. Cloning 1Dx5 Gene from the Xinjiang Wheat Variety and Constructing Its Expression Vector,S512.1
  6. Mitochondrial Genome Sequence Analysis of a Brassica Juncea Landrace,S565.4
  7. Molecular Epidemiology about White Spot Syndrome in Jiangsu Province,S945.1
  8. Isolation and Identification of NDV Local Epidemic Strains and the Comparison of Vaccination Programs,S852.65
  9. Establishment of a Nested PCR Assay and Analysis of the Full Genome of HUMAN COSAVIRUS,S855.3
  10. Cloning, Expression and Enzymatic Characterization of Two Malate Dehydrogenases,Q946.5
  11. Cloning and Sequence Analyses of CONSTAN (CO) and LEAFY (LFY) Homologous Gene of Dendrocalamus Xishuangbannaensis,Q943.2
  12. The Fusion Expression of a Pea Actin Isoform (PEAc3) and Green Fluorescent Protein (GFP) and the Biochemical Characteristics Analysis to the Fusion Protein,Q946.1
  13. Sequences Variation of Mitochondrial 12S rRNA Gene and Molecular Phylogeny of 20 Species Birds(AVES: Charadriiformes),Q958
  14. Sequence Analysis of Mitochondrial DNA on Hyriopsis Schlegelii,S917.4
  15. Ae.tauschii gliadin gene cloning and sequence analysis,S512.1
  16. Preliminary Study on the Molecular Mechanism of Self-incompatibility in Brassica Napus L,S565.4
  17. Molecular Genetic Analyses of a Family with Congenital Cataract,R776.1
  18. Confirmation of the G3635A Mutation in the Mitochondrial ND1 Gene as Causative in Leber’s Hereditary Optic Neuropathy,R774.6
  19. Arbovirus Detection and Gene Sequence Analysis of Viral Encephalitis and Unknown Fever Patients in Yunnan Province,R512.3
  20. Study on the Expression of Partial Different Genes in Segments about Moniezia,Q78
  21. Evolutionary analysis of equine infectious anemia virus part attenuated strains in vitro and in vivo,S852.65

CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Horticultural Crops Pest and Disease Control > Vegetable pests > Melons, pests and diseases > Cucumber pests and diseases
© 2012 www.DissertationTopic.Net  Mobile