Agrobacterium-mediated transformation of alfalfa (Medicago sativa) using a synthetic cry3a gene to enhance resistance against alfalfa weevil View Full Text


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Article Info

DATE

2012-11-25

AUTHORS

Masoud Tohidfar, Naser Zare, Gholamreza Salehi Jouzani, Seide Maryam Eftekhari

ABSTRACT

To introduce genetic resistance against alfalfa weevil (Hypera postica), leaves and petiole explants of three commercial alfalfa genotypes, including Km-27, Kk-14 and Syn-18 were transformed with Agrobacterium tumefaciens strains GV101, LBA4404 and AGL01. All the Agrobacterium strains used harbored the recombinant binary vector pBI121 containing a synthetic cry3a gene under the control of CaMV35S promoter as well as the nptII gene as selectable marker. Transformed explants were cultured on callus-induction medium, and the germinated somatic embryos were then transferred to the regeneration medium. The primary transformants were evaluated by PCR and Southern blot analysis. The results indicated successful integration of the target gene into the genomes of primary transgenic lines. Moreover, the expression of Cry3a protein in the transgenic plants was confirmed by ELISA method. Three transgenic lines, including TL6, TL8 and TL11 showed significantly higher levels of insect resistance against H. postica larvae (mortality rate of 73–90 % after infestation), in comparison with the control plants during the two-year bioassays. All transgenic plants were fertile and no irregular behavior in terms of growth and the morphological traits were observed. Transgenic plants developed during the course of this study are currently being grown in greenhouse and will be crossed with each other for seed production. More... »

PAGES

227-235

References to SciGraph publications

  • 2011-09-13. Transgenic expression of phytase and acid phosphatase genes in alfalfa (Medicagosativa) leads to improved phosphate uptake in natural soils in MOLECULAR BREEDING
  • 2009-11-03. Stress-inducible expression of GmDREB1 conferred salt tolerance in transgenic alfalfa in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 1997-10. Enhanced resistance to two stem borers in an aromatic rice containing a synthetic cryIA(b) gene in MOLECULAR BREEDING
  • 2008-10-14. Transgenic rice expressing Allium sativumleaf agglutinin (ASAL) exhibits high-level resistance against major sap-sucking pests in BMC PLANT BIOLOGY
  • 2008-02-14. Effect of proteinase inhibitors on Indian alfalfa weevil (Hypera postica Gyll.) growth and development in ACTA PHYSIOLOGIAE PLANTARUM
  • 2005-02. Transgenic alfalfa secretes a fungal endochitinase protein to the rhizosphere in PLANT AND SOIL
  • 2005-02. Comparative study of the expression of the native, modified, and hybrid cry3a genes of Bacillus thuringiensis in prokaryotic and eukaryotic cells in RUSSIAN JOURNAL OF GENETICS
  • 1995-12. Strain specificity in transformation of alfalfa by Agrobacterium tumefaciens in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 1995-02. Production and field performance of transgenic alfalfa (Medicago sativa L.) expressing alpha-amylase and manganese-dependent lignin peroxidase in EUPHYTICA
  • 1995-08. Genetic transformation of commercial breeding lines of alfalfa (Medicago sativa) in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2012-03-13. Expression of a recombinant human erythropoietin in suspension cell cultures of Arabidopsis, tobacco and Medicago in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2001-06. A rapid and high yielding DNA miniprep for cotton (Gossypium spp.) in PLANT MOLECULAR BIOLOGY REPORTER
  • 2007-12-19. Expression of hybrid cry3aM–licBM2 genes in transgenic potatoes (Solanum tuberosum) in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2011-09-22. Physiological responses to salt stress of T2 alfalfa progenies carrying a transgene for betaine aldehyde dehydrogenase in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 2001-07. Single-copy primary transformants of maize obtained through the co-introduction of a recombinase-expressing construct in PLANT MOLECULAR BIOLOGY
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