Agrobacterium -mediated transformation of cotton (Gossypium hirsutum) using a heterologous bean chitinase gene View Full Text


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Article Info

DATE

2005-10

AUTHORS

Masoud Tohidfar, Mojtaba Mohammadi, Behzad Ghareyazie

ABSTRACT

Cotton (Gossypium hirsutum L., var. Coker 312) hypocotyl explants were transformed with three strains of Agrobacterium tumefaciens, LBA4404, EHA101 and C58, each harboring the recombinant binary vector pBI121 containing the chi gene insert and neomycin phosphotransferase (nptII) gene, as selectable marker. Inoculated tissue sections were placed onto cotton co-cultivation medium. Transformed calli were selected on MS medium containing 50 mg l−1 kanamycin and 200 mg l−1 cepotaxime. Putative calli were subsequently regenerated into cotton plantlets expressing both the kanamycin resistance gene and βglucuronidase (gus) as a reporter gene. Polymerase chain reaction was used to confirm the integration of chi and nptII transgenes in the T1 plants genome. Integration of chi gene into the genome of putative transgenic was further confirmed by Southern blot analysis. ‘Western’ immunoblot analysis of leaves isolated from T0 transformants and progeny plants (T1) revealed the presence of an immunoreactive band with MW of approximately 31 kDa in transgenic cotton lines using anti-chitinase-I polyclonal anti-serum. Untransformed control and one transgenic line did not show such an immunoreactive band. Chitinase specific activity in leaf tissues of transgenic lines was several folds greater than that of untransformed cotton. Crude leaf extracts from transgenic lines showed in vitro inhibitory activity against Verticillium dahliae.Transgenic plants currently growing in a greenhouse and will be bioassayed for improved resistance against V. dahlia the causal against of verticilliosis in cotton. More... »

PAGES

83-96

References to SciGraph publications

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  • 1999-02. Transformation of a Texas cotton cultivar by using Agrobacterium and the shoot apex in THEORETICAL AND APPLIED GENETICS
  • 1997-04. A fungal chitinase gene fromRhizopus oligosporus confers antifungal activity to transgenic tobacco in PLANT CELL REPORTS
  • 1983-02. Chitinase in bean leaves: induction by ethylene, purification, properties, and possible function in PLANTA
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  • 2000-04. Susceptibility to Agrobacterium tumefaciens and cotyledonary node transformation in short-season soybean in PLANT CELL REPORTS
  • 1997-09. Heterologous chitinase gene expression to improve plant defense against phytopathogenic fungi in JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
  • 1987-03. Transformation of cotton (Gossypium hirsutum L.) by Agrobacterium tumefaciens and regeneration of transgenic plants in PLANT MOLECULAR BIOLOGY
  • 1996-06. Transgenic plant production mediated by Agrobacterium in Indica rice in PLANT CELL REPORTS
  • 1984-06. Presence of a plant cell wall is not required for transformation of Nicotiana by Agrobacterium tumefaciens in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 1996-04. Transformation of pickling cucumber with chitinase-encoding genes using Agrobacterium tumefaciens in PLANT CELL REPORTS
  • 2000-06. Transgenic grapevine plants expressing a rice chitinase with enhanced resistance to fungal pathogens in PLANT CELL REPORTS
  • 2001-06. A rapid and high yielding DNA miniprep for cotton (Gossypium spp.) in PLANT MOLECULAR BIOLOGY REPORTER
  • 1987-03. Genetically Transformed Cotton (Gossypium Hirsutum L.) Plants in NATURE BIOTECHNOLOGY
  • 1970-08. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 in NATURE
  • 1995-07-01. Genetic Engineering of Rice for Resistance to Sheath Blight in NATURE BIOTECHNOLOGY
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