Dissertation > Excellent graduate degree dissertation topics show

Genetic Transformation and Molecular Analysis of Transgenic Stem Mustard Plants Coexpressing Coat Protein Genes of TuMV and CMV

Author: ZhangWei
Tutor: GuoDePing；ChenJiShuang
School: Zhejiang University
Course: Olericulture
Keywords: stem mustard cotyledon turnip mosaic virus cucumber mosaic virus transformation
CLC: S637.3
Type: Master's thesis
Year: 2006
Downloads: 58
Quote: 0
Read: Download Dissertation

Abstract


A transformation system of TuMV CP and CMV CP gene mediated by Agrobacterium tumefaciens was established in stem mustard (Brassica juncea var. tsatsa). Cotyledons of stem mustard were used as the explant, and transformants were successfully obtained in the present experiment. This study provided theoretical and technical basis for obtaining virus-resistant plants by modern molecular biotechnology and plant genetic engineering, and technical platform for gene transformation in mustard and Cruciferae as well. The resultes are as follows: 1. Construction of plant bivalent expression vector of TuMV CP and CMV CP genesThe specific primers were designed and synthesized according to the published sequence of TuMV CP and CMV CP gene, TuMV CP and CMV CP gene fragmentes were obtained by PCR amplification using pMD-TuMV CP and pUC-CMV CP as templates, and the TuMV CP amplified fragment was cloned into pCambial301 vector to produce recombinant clone pCam-TuMV CP, and CMV CP amplified fragment was cloned into pBI121 vector to produce recombinant clone pBI-CMVCP.containing CMV CP gene fragment was obtained by digesting the pBI-CMV CP vector with EcoRI and PstI,we could get Plant Bivalent Expression Vector of TuMV CP and CMV CP gene through containing CMV CP gene fragmentinserted into pCam-TuMV CP. pCam-TuMV CP-CMV CP was transformed into A. tumefaciens EHA105 by freezing and thawing method.The positive clones confirmed by PCR and digestion were used for transformation of stem mustard (Brassica juncea var. tsatsd).2. Establish of transformation system using cotyledon as explantThe effect of different concentration of antibiotics on regeneration of cotyledonswas studied. Meanwhile, the effect of pre-culture time(ld, 2d? 3d, 4d),co-culturetime(ld> 2d, 3d. 4d) and on transformation efficiency of cotyledon were investigated in the present study. The best transformation frequency was found in 2-day pre-cultured explants and 10-15 min infection after 2-day co-culture through 12.5mg/l Hgy and 500 mg/1 Carb differentiation medium at 28 °C.3. Molecular identification of transformed shootsBy PCR and RT-PCR analysis, we found that TuMV CP and CMV CP gene were inserted into the stem mustard genome and expressed successfully

Related Dissertations

  1. Rebellion of an and Shi and Poetic Style Evolution from Kaiyuan to Tianbao,I207.22
  2. Research of Wavefront Transformation Based on Liquid Crystal Spatial Light Modulator,TN761
  3. Service Model Driven Architecture (SMDA) Oriented Modeling Techniques and Tool Implemention,TP311.52
  4. Rural Energy Transformation and the Benefits of Ecological Environmental,S210.3
  5. Improvement of the Resistance of Nicotiana Tabaccum to Virus by RNAi and Initial Establishment of Genetic Transformation System of L.regale,S435.72
  6. Cloning 1Dx5 Gene from the Xinjiang Wheat Variety and Constructing Its Expression Vector,S512.1
  7. Study on Transformation of Cut Chrysanthmemum with DdICE1 Gene,S682.11
  8. Study on Agrobacterium-Mediated Transformation of ’Benihonpe’ Strawberry with ALA Synthase Gene,S668.4
  9. Studies on Aphid Resistance Identification, Resistance Related Mechanisms and LLA Gene Transformation of Cut Chrysanthemum,S682.11
  10. Functional Analysis of Solanum Torvum StPGIP Gene Related to Verticillium Resistance,S572
  11. Genetic Transformation of Potato with 3GT Gene,S532
  12. Cysteamine on T Lymphocyte Proliferation in Post-Operation Goats and Its Mechanism Underlying,S858.27
  13. China’s Third since the Social Transformation of Competitive Sports Development Path,G812.0
  14. Transformation of Tobacco with Arabidopsis Thaliana MAP65-6 and TUA-6,Q943.2
  15. Study on Management Performance of Enterprise Based on the Transformation of Value-added Tax,F224.32
  16. Local area WGS-84, Beijing 54 and 80 Xi'an coordinate conversion application,P226.3
  17. Research on Topology of Improved Unified Power Quality Conditioner and Control Strategy,TM761
  18. High-frequency Plant Regeneration and Genetic Transformation of Jatropha Curcas L.,S792.99
  19. Absorption and Transformation,I207.42
  20. Developing Second Best Institutions: A Competition Policy That Fits China’s Mode of Development,D922.294
  21. Classification Research Based on Evidence Theory and Covering Rough Sets,TP18

CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Mustard > Stem mustard
© 2012 www.DissertationTopic.Net  Mobile