Paper Information

Journal:   IRANIAN JOURNAL OF AGRICULTURAL SCIENCES (JOURNAL OF AGRICULTURE)   2006 , Volume 37-1 , Number 3 (AGRONOMY & CROP BIOTECHNOLOGY); Page(s) 539 To 544.
 
Paper: 

A STUDY OF GENE TRANSFORMATION BY VACUUM INFILTRATION INTO RAPESEED AND ARABIDOPSIS PLANTS

 
 
Author(s):  GHORBANI MESHKIN M., JALALI JAVARAN MOKHTAR, SEPAHVAND N.A.
 
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Abstract: 

Different gene transfer strategies, based on tissue culture, are employed in plants such as rapeseed. These strategies suffer from constraints like; need of transgenic cell regeneration, somaclonal variation, in ad9ition to being costly and time consuming. Considering the constraints, researchers often resort to planta transformation. This strategy, in addition to simplicity, low cost, and avoidance of somaclonal variation is of the feasibility for species for which tissue culture application hasn't been developed. Vacuum infiltration was primarily applied in Arabidopsis thaliana L. It has been used in several other species like radish. This investigation explains gene transfer into rapeseed using vacuum infiltration, for the first time in Iran. An arabidopsis inbred line was used as a check, and the frequencies of transgenic plants were compared. Two Agrobacterium strains, EHA101 and C58(pGV3101) containing pVW295 plasmid having NPTII and GDS genes were used in the investigation. The flowering rapeseed and arabidopsis plants were inverted into the agrobacterium culture in a vacuum chamber applying -350 to -400 mmHg of pressure for 30-40 seconds. Seeds from infiltrated rapeseed and arabidopsis plants were selected in media containing 50 and 30 mgL-1 Kanamycin respectively. To confirm the GUS gene transfer into the selected progenies histochemical evaluation was conducted. The histochemical assay confirmed the GDS gene transfer. Evaluation of the infiltrated plant seeds revealed that the frequency of transgenic arabidosis was more than that in rapeseed. Agrobacterium strain EHA101 was more efficient than C58 (pGV3101) in gene transfer into either of arabidosisor rapeseed.

 
Keyword(s): GENE TRANSFORMATION, ARABIDOPSIS, BRASSICA NAPUS, VACUUM INFILTRATION, GDS GENE
 
References: 
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