Dissertation > Excellent graduate degree dissertation topics show

Studies on Construction of Regeneration System and Genetic Transformation of Puccinellia Chinampoensis

Author: WangTao
Tutor: LiuCanKui
School: Northeast Forestry University
Course: Biochemistry and Molecular Biology
Keywords: Puccinellia chinampoensis embryo callus Agrobacter-ium tumefaciens genetic transformation tissue browning
CLC: S543.9
Type: Master's thesis
Year: 2011
Downloads: 46
Quote: 0
Read: Download Dissertation

Abstract


Puccinellia chinampoensis is a special gramineous plant like rice and wheat, which can survive well in the saline and alkali environment with severe drought and salt stress. The difference from rice or wheat is Puccinellia chinampoensis has evolved a series of resistance mechanism via years of adaptation in the saline and alkali soil. In this study, transformation system of Puccinellia chinampoensis has been constructed and optimized, following with optimized regeneration system, through plant tissue culture technique. The whole constructed system will be pre-determinative for the further studies as gene interference.In this study, some crucial factors which affected the embryo culturing using Puccinellia chinampoensis as the main plant material were discussed comprehensively and systemically. And the genetic transformation of put-LS2 mediated by Agrobacterium tumefaciens was followingly conducted using the above callus. The main results obtained in this study contains as follows.In the callus induction process of Puccinellia embryo, different sterilant and sterilized time to seed embryo showed different effects and induced situation obviously. In this study, the best combination was 70% ethanol for 3min and 10% sodium hypochlotous for 30min. The rate of callus induction was different in the medium containing different concentrations of 2,4-D and the optimized was 4 mg/L. The altered medium I (N6max+MSmin+2×MSFe+5×MSvitamin (inositol no change)) was chosen to be the basic medium to induce the callus. The time of cultivation in light also have effect on the callus induction. The shorter of the light time, the higher rate of the callus induction would get.In the callus subculture of Puccinellia chinampoensis seeds,different mediums showed different impacts. The result showed that the modified medium S(MSmax+4×MSmin+MSFe+ N6vitamin) was chosen as the basic subculture medium.1mg/Lphytohormone ABA was added to the medium to improve the quality and the quantity of embryo callus.In the differentiation of callus of Puccinellia chinampoensis embryo, MS was the differentiation medium. Different ratio of hormone had different effects on differentiation. The result indicated that the best was 6-BA 0.4mg/L+IAA 0.04mg/L.Sucrose was substituted with sorbital and maltose, which could inhibit callus browning obviously and increase differentiation rate.500mg/L proline was added into induction medium, succeeded medium and differentiation medium to effectively prevent callus from browning and to improve differentiation rate.In the transformation test of Puccinellia chinampoensis callus, using kanamycin (Kana) to screen transformed callus, the optimized concentration was 50mg/L. The beat way of remove of bacterial is filtration. pBl121-GUS was transformed into the callus mediated by Agrobacter-ium tumefaciens, and the transformation rate was 47.14%.

Related Dissertations

  1. Improvement of the Resistance of Nicotiana Tabaccum to Virus by RNAi and Initial Establishment of Genetic Transformation System of L.regale,S435.72
  2. Biological Effect of Rare Earth Element and Genetic Transformation on Arnebia Euchroma(Royle) Johnst.Cell,S567.239
  3. The Genetic Transformation and Functional Analysis of GmFtsH,S565.1
  4. Cloning of Proteinase Inhibitor Gene CYS and API from Wild Potatoes and Preliminary Study of Potato and Tobacco Transformation with Spcys,S532
  5. Study on Agrobacterium-Mediated Transformation of ’Benihonpe’ Strawberry with ALA Synthase Gene,S668.4
  6. AttM Transgenic Colored Calla Lily and Antibodies Preparation of Ahl-Degrading Enzyme AttM,S682.264
  7. Studies on Agrobacterium-midiated Genetic Transformation in Brassica Campestris Subsp. Chinensis Var. Parachinensis,S634.5
  8. Functional Analysis of Solanum Torvum StPGIP Gene Related to Verticillium Resistance,S572
  9. Construction of RNA Interference Vectors of Pin Genes Determining Grain Hardness and Genetic Transformation in Wheat,S512.1
  10. Lisianthus ACC synthase gene RNA interference,S682.19
  11. Optimization of Regeneration System of Sea Island Cotton and Study on the Genetic Transformation of NAC Transcripts Family Gene,S562
  12. Establishment of Technique System of Agrobacterium-mediated Transformation Cotton (Gossypium Barbadense L.) and Identification of Resistance to Cotton Boll Worm of Bt Transgenic Cotton,S562
  13. Preliminary Functional Study of Rice OsLIR1 Gene,S511
  14. Agrobacterium Rhizogenes Mediated Bt of Cry8e Transformation of Soybean Resistance to the Holotrichia,S565.1
  15. Studies on Gene Clone and Functional Analysis of a Flavonoid 3-O-glucosyltransferase Gh3GT from Natural Brown Cotton Fiber,S562
  16. Establishment of T-DNA Insertion Mutation Library of Fusarium Oxysporum f.sp.cubense Race1 and Location of Insertion Site of Some Related Pathogenicity Mutants,S436.68
  17. PGIP Cloning and Antifungal Activity,S432.44
  18. Optimization of Genetic Transformation System and Preliminary Investigation on the Infection Mechanism of Ustilago Hordei,S435.123
  19. Study on Transformation of Papaya Ringspot Vir (PRSV) ci and nib Gene Inverted Repeats Experssion Vector,S436.67
  20. Optimization of Regeneration System of Tissue Culture and Transformation of KN2 Gene in Wheat,S512.1
  21. The Research of the Role of EDR1,EDR2 in Plant Resistance,S435.111

CLC: > Agricultural Sciences > Crop > Fodder crops,pasture > Perennial grass forages > Other
© 2012 www.DissertationTopic.Net  Mobile