Dissertation > Excellent graduate degree dissertation topics show

Functional Analysis of an Sclerotinia Disease Resistance Gene

Author: WangWeiQing
Tutor: WangXinYu
School: Nanjing Agricultural College
Course: Biochemistry and Molecular Biology
Keywords: Sclerotinia sclerotiorum Stellaria apetala ucria G-protein-coupled receptor Agrobacterium-medialed transformation Brassica napus Nicotiana tabacum
CLC: S435.654
Type: Master's thesis
Year: 2011
Downloads: 13
Quote: 0
Read: Download Dissertation


Oilseed rape (Brassica napus) is an important industrial crop and a major oil crop. It plays important roles in national economy and agricultural production. Sclerotinia stem rot caused by Sclerotinia sclerotiorum is the most devastating disease of oilseed rape in China. To date, no complete resistance has been found in rapeseed germlpasm and its relatives. Stellaria apetala ucria is a common weed in the field of canola, it has non-host resistance to Sclerotinia sclerotiorum. It is of very significance to understand the mechanisms from molecular level of the interactions between Brassica napus and Sclerotinia sclerotiorum and to isolate genes associated with susceptibility and resistance to the infection by Sclerotinia sclerotiorum, which should be helpful to protect plant from infection. Sspgld, one of endopolygalacturonases, is an important fungal effector secreted by the necrotrophic fungus Sclerotinia sclerotiorum during early infection. Using sspgld as bait, a membrane protein named ppn20 was identified by yeast two-hybrid screening of cDNA library of Stellaria apetala ucria. In previous study, we have demonstrated that ppn20 is a G protein-coupled receptor, and ppn20 transcription in Stellaria apetala leaf was induced by Sclerotinia sclerotioru. In this study, we want to further make clear its function. We use Agrobacterium-mediated transformation to obtain Brassica napus and Nicotiana Tabacum transgenic plants. Kanamicin resistance, PCR and RT-PCR were used to screen transgenic plants. The phenotype was analysed by inoculation with Sclerotinia sclerotiorum. Thus, plant expression vector ppn20-pBinplus was transformed into Agrobacterium. Agrobacterium-medialed transformation was used to transform Brassica napus and Nicotiana Tabacum. In total,10 positive Brassica napus plants were obtained by screening with kanamicin, of which 7 plants survived. Ppn20 can be detected in all of these positive plants by PCR and RT-PCR analysis. The resistance phenotype of these plants was assayed by inoculation with Sclerotinia sclerotiorum, the results showed that both of the control plants and the transgenic plants were easily infected with Sclerotinia sclerotiorum. But the fungus was highly restricted in the infection site of the transgenic Nicotiana tabacum, some plants even could not be infected by the fungus. Ppn20 was constructed in plant expression vector PVX, and which was transformed into Agrobacterium. Arobacterium infiltration assay was used to transiently express the ppn20 gene in N. benthamiana leaves. The resistance phenotype of the infiltration area was assayed by inoculating with Sclerotinia sclerotiorum, the results showed that the fungus has not restricted in the sites where ppn20 was infiltrated. Mating-based split-ubiquitin systems in yeast cells proved that there is no interaction between membrane protein ppn20 and the G protein of canola. This may be the reason that ppn20 gene does not show resistance to Sclerotinia sclerotiorum in Brassica napus.

Related Dissertations

  1. Identification of Cyto-and Molecular Genetics of Multisomic Addition Line "Nj08-063" in Brassica Napus and Its Agronomical Traits,S565.4
  2. Cloning Andanalysis of NAC Transcription Factor Gene in Tobacco(Nicotiana Tabacum L.),Q943.2
  3. Preliminary Studies on 4 EMS-Induced Mutants in Brassica Napus L,S565.4
  4. Studies on Extracting Mitochondrial DNA and Gene Expressions in Brassica Napus L,S565.4
  5. Studies on Screening in Progeny Population of Distant Hybridization between Brassica Napus L. and Descurainia Sophia,S565.4
  6. Preliminary Studies on Constructing Brassica Napus L Mutant Library by EMS Inducement,S565.4
  7. Anatomy of Root, Stem and Its Lignin Content and Expression of Related Key Genes in Lignin Synthesis of Lodging Resistant Brassica Napus L.,S565.4
  8. Analysis of Difference in Oil and Fatty Acids Accumulation in Developing Seeds of Brassica Napus L. from Various Altitude Localities,S565.4
  9. Salt-Tolerance Evaluation of Brassica Napus Germplasm in the Bud and Seeding and Establishment of the Two-Dimensional Electrophoresis System,S565.4
  10. Study on Fitness of Backcross Generations between Glyphosate-Resistant Transgenic Oilseed Rape and Four Geographic Populations of Wild Brassica Juncea,S565.4
  11. Effects and Mechanism of Exasa on the Growth of Rapeseed Exposed to CD,S565.4
  12. Cloning and Expressing of Lysophosphatidic Acid Acyltransferase Gene from Brassica Napus L,S565.4
  13. Ning RS-1 SSH cDNA library construction and PGIP gene and its expression vector,S565.4
  14. DES and INSL3 on cultured mouse gubernaculum testis cells LGR8 transcription and expression,R726.9
  15. Linkage Map Construction and Identification of Quantivitive Trait Loci (QTL) for Oil Content and Related Traits in Brasscia Napus,S565.4
  16. Preliminary Study on the Molecular Mechanism of Self-incompatibility in Brassica Napus L,S565.4
  17. Studies on Core SSR Primers for Brassica Napus,S565.4
  18. Comparative Analysis of the Genomic Region Containing Recessive Genic Male Sterility Gene in Rapeseed and Development of Sequence-specific Markers,S565.4
  19. Additional lines of Brassica napus and Brassica Genome A study interspecific hybridization,S634
  20. Identification of Molecular Markers and Preliminary Mapping of Fertility Restorer Gene (Rfp) for the ’Polima’ CMS in Brassica Napus L.,S565.4

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