Irreproducibility of the soybean pollen-tube pathway transformation procedure |
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Authors: | Huixia?Shou Reid?G?Palmer Email author" target="_blank">Kan?WangEmail author |
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Institution: | (1) Department of Agronomy, Iowa State University, 50011-1010 Ames, IA, USA;(2) Agricultural Research Service and Corn Insects and Corn Genetics Research Unit, USDA, USA;(3) Department of Agronomy, Iowa State University, 50011-1010 Ames, IA, USA;(4) Department of Zoology/Genetics, Iowa State University, 50011-1010 Ames, IA, USA |
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Abstract: | The interest in developing tissue culture-independent genetic transformation methods for plants has been growth. The pollen-tube
pathway transformation technique is one method; however, this method is controversial because it is difficult to duplicate
and produces insufficient molecular evidence to confirm transformation. Our objective was to evaluate the robustness of the
soybean pollen-tube pathway technique (Glycine max L. Merr.). Solutions of purified DNA constructs carrying abar marker gene and agus reporter gene or a gene of interest (npk1) were applied to severed styles of flowers 6–8 h after self-pollination. The experiment was repeated 3 summers in the field,
in which 4 DNA constructs and 7 soybean genotypes were tested. A total of 4793 progeny seeds were harvested from 5590 individually
treated soybean flowers. All seeds were germinated and screened for transformants with herbicide spray, histochemical GUS
assay, and Southern blot analysis. Although 2% of progenies showed partial resistance to the herbicide, no positive plants
were identified from GUS assay and Southern analysis. Our results indicate that soybean pollen-tube pathway transformation
is not reproducible. |
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Keywords: | pollen-tube pathway soybean transformation |
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